Skip to main content
NCBI home page
ACS Omega logoLink to ACS Omega
. 2026 Jul 27;11(31):47148–47161. doi: 10.1021/acsomega.6c07405

Folate-Targeted Co-Delivery of Curcumin and siRela Nanoparticles Synergistically Treat Sepsis-Associated Acute Kidney Injury

Xiangyi Xu †,‡,§, Jingjin Tao †,‡,§, Zhongjun Shen †,‡,§, Qian Zhang ‡,§, He Wang ‡,§, Yuying Nie ‡,§, Huike Guo †,‡,§, Qi Liu ‡,§, Zhongxin Li ‡,§, Chong Wang ‡,§, Donglin Liu ‡,§, Jiachen Pei ‡,§, Chengqian Ma †,‡,§, Shuo Yang ‡,§,*, Liyan Cui †,‡,§,*
PMCID: PMC13470973  PMID: 42597958

Abstract

The increasing burden of Acute Kidney Injury (AKI) significantly impacts human health. Approximately 70% of septic patients develop AKI, which markedly increases the risk of in-hospital mortality. However, current treatment for Sepsis-Associated Acute Kidney Injury (SA-AKI) is mainly supportive, and no specific therapy is available. siRNA therapy is an emerging approach that inhibits mRNA of specific targets to exert therapeutic effects. Based on the gene silencing principle of siRNA, we developed a targeted bifunctional coloaded nanoparticle, PDA@Cur–PEI-PEG-FA/siRela (PCPPFS), using folic acid (FA) as a targeting moiety. In this system, curcumin acts as a therapeutic agent and synergizes with siRela to exert anti-inflammatory and antioxidative stress effects, thereby reducing apoptosis and pyroptosis of renal tubular epithelial cells. This nanosystem demonstrated good targeting and therapeutic effects in both in vitro and in vivo experiments, providing a new approach for SA-AKI therapy.

graphic file with name ao6c07405_0010.jpg

graphic file with name ao6c07405_0008.jpg

1. Introduction

Acute Kidney Injury (AKI) is defined as an increase in serum creatinine level by at least 50% within 7 days, or an increase of at least 0.3 mg/dL within 2 days, or a significant reduction in urine output within 6 h. More than 10–15% of hospitalized patients experience AKI, and this proportion can reach 30–60% in intensive care units. The growing burden of AKI seriously affects people’s lives. Sepsis is a life-threatening organ dysfunction caused by a dysregulated host response to infection. SA-AKI requires meeting the diagnostic criteria for both sepsis and AKI. About 70% of septic patients develop AKI, and the condition often progresses to stage 3 AKI. Moreover, AKI significantly increases the risk of in-hospital mortality and the likelihood of developing chronic kidney disease. Notably, patients with AKI caused by bacterial infection have a higher risk of in-hospital death than those with AKI from other causes. The pathological mechanisms of SA-AKI are complex and may involve systemic and renal inflammation, oxidative stress, various forms of cell death (such as apoptosis, pyroptosis, and ferroptosis), microcirculatory and endothelial dysfunction, and metabolic reprogramming. They are not yet fully understood. Currently, treatment for SA-AKI is mainly supportive, with no effective prevention or specific therapeutic strategy. Therefore, developing specific treatments is of great significance. −

In septic AKI, the inflammatory response of renal tubular epithelial cells is a key pathological mechanism. Rela (p65) is the core functional subunit of the NF-κB pathway and a crucial regulator of renal inflammation and apoptosis in septic AKI. When LPS stimulates renal tubular epithelial cells, it binds to Toll-like receptor 4 (TLR4), thereby activating the NF-κB signaling pathway. Activated Rela acts as a transcription factor, promoting the production of pro-inflammatory cytokines such as IL-6, which activates inflammasomes, triggers a renal inflammatory storm, and exacerbates tubular damage and renal dysfunction. Excessive NF-κB activation is an important pathogenic mechanism of LPS-induced septic AKI, and inhibiting NF-κB activation significantly protects the kidneys and alleviates SA-AKI. − Therefore, inhibiting Rela expression is a promising therapeutic strategy for AKI.

Curcumin is a natural phenolic compound extracted from the rhizome of turmeric. Studies have shown it to be a potential kidney-protective agent. − Curcumin reduces the expression of various inflammatory factors, such as IL-1β and IL-6, thereby alleviating renal inflammatory damage. Curcumin also prevents renal tubular oxidative damage and activates Nrf2, HO-1, and other activities, thus reducing ROS production. Furthermore, curcumin restores mitochondrial homeostasis and reduces apoptosis, pyroptosis, and ferroptosis. − In septic AKI, curcumin exerts protective and therapeutic effects, lowering the levels of kidney injury markers and improving renal function. − However, the administration efficiency and bioavailability of curcumin are very low, necessitating suitable carriers to improve its absorption. Nanomaterials can improve the bioavailability of curcumin, enhance targeting, and reduce toxicity, making them advantageous dosage forms for curcumin delivery. ,

siRNA therapy is an emerging therapeutic approach. In 2018, the first siRNA drug, Patisiran, was approved in the United States and Europe, marking the official beginning of the RNAi therapy era. siRNAs are approximately 20–25 nucleotides in length. They form the RNA-induced silencing complex (RISC) inside cells. RISC binds to the mRNA of a specific target gene, leading to mRNA degradation and preventing normal protein translation, thus reducing the expression of a specific protein. This gene silencing effect can theoretically inhibit almost any target, making it a highly promising novel therapy. However, siRNA has low cellular uptake, is susceptible to degradation by RNases, and has poor stability in serum, with a half-life of approximately 5 min. Therefore, developing a carrier with good biosafety that can stabilize siRNA delivery and uptake is crucial for siRNA administration. −

Active targeting strategies involve modifying and designing nanodelivery vehicles, coupling them with moieties that enable active targeting of specific organs or cells. Folate receptors are highly expressed on renal tubular epithelial cells; thus, folic acid can serve as a targeting group to actively target these cells. Polydopamine (PDA) is produced by the self-polymerization of dopamine hydrochloride. It has good biocompatibility, can carry drugs into cells, and possesses abundant active sites for binding various drugs or targeting groups. Studies have shown that PDA nanoparticles degrade more readily under ROS-rich conditions, promoting drug release in tissues with intense inflammatory reactions. PEI is a cationic polymer and a traditional, potent material for loading siRNA. Modification with PEG can improve the biosafety of PEI and prolong the circulation of nanomedicines in vivo. ,

This study aimed to target renal tubular epithelial cells that highly express folate receptors using folic acid as a targeting moiety, delivering siRela and curcumin via PDA and PEI. Our results demonstrate effective Rela knockdown in vitro, along with antiapoptotic, antioxidative stress, and anti-inflammatory effects. In a CLP mouse model of sepsis, this nanomedicine effectively knocked down Rela expression and improved renal function, representing a potential treatment for SA-AKI (Figure ).

1.

1

Open in a new tab

Schematic illustration of PCPPFS nanoparticle synthesis and therapeutic effect in mice. PDA was generated by self-polymerization of dopamine hydrochloride, followed by sequential modification with curcumin, PEI, PEG, FA, and siRela. After intravenous injection into mice, the FA moiety of PCPPFS nanoparticles actively targets folate receptors highly expressed on renal tubular epithelial cells. Following cellular internalization via endocytosis, curcumin and siRela work together to exert therapeutic effects, including antioxidation, reduction of inflammation, antipyroptosis, and antiapoptosis, thereby improving kidney injury markers. Some of the images were created by biogdp.com.

2. Materials and Methods

2.1. Materials

LPS (L2630) was provided by Sigma-Aldrich (St. Louis, MO, USA). Anti-β-Actin (A0101) was obtained from Lablead. Anti-NGAL (26991–1-AP) was purchased from Proteintech. Anti-Rela (D14E12) was obtained from Cell Signaling Technology. Anti-GSDMD (PU224937S) was from Abmart. HRP-conjugated goat antimouse IgG (H+L) secondary antibody (S0100) and HRP-conjugated goat antirabbit IgG (H+L) secondary antibody (S0101) were purchased from Lablead.

N-Hydroxysuccinimide (CAS# 6066–82–6, 1037423) and 1-Ethyl-(3-(dimethylamino)­propyl)­carbodiimide hydrochloride (CAS# 25952–53–8, 1027609) were obtained from Leyan. Folic acid (CAS# 59–30–3, F809516) and branched polyethylenimine (CAS# 25987–06–8, P766520) were obtained from MACKLIN. NH2–PEG-COOH (N861524), Ammonium hydroxide solution (CAS# 1336–21–6, A834475), and Dopamine hydrochloride (CAS# 62–31–7, D756911) were from MACKLIN. Curcumin (CAS# 458–37–7, C400271) was purchased from Aladdin. siRNA and Cy5.5-labeled siRNA were purchased from hippobio, with sequences listed in Table S1.

2.2. Synthesis of PCPPFS

First, polydopamine (PDA) nanoparticles were synthesized using an alkali-catalyzed self-polymerization method. Dopamine hydrochloride (50 mg/mL) was added dropwise to a mixed solution containing aqueous ammonia (3.5 mL), anhydrous ethanol (40 mL), and distilled water (90 mL), and the reaction was stirred at room temperature for 24 h. During this process, the reaction solution gradually turned brownish-yellow, as dopamine underwent oxidative self-polymerization under alkaline conditions to form PDA nanoparticles with catechol and quinone groups via covalent and noncovalent interactions. The reaction mixture was centrifuged four times (13,000 rpm, 40 min, 4 °C), and the pellet was resuspended in distilled water to obtain purified PDA nanoparticles. The concentration was determined by dry weight. Subsequently, a curcumin (Cur) ethanol solution (2 mg/mL, 10 mL) was added to the PDA dispersion and stirred at room temperature for 24 h. Curcumin was loaded onto the PDA surface through hydrophobic interactions, π–π stacking, etc., forming PDA@Cur nanoparticles (PC). After four washes by centrifugation (13,000 rpm, 40 min, 4 °C), an aqueous solution of branched polyethylenimine (PEI) was added dropwise to the PDA@Cur dispersion and reacted for 2 h at room temperature. The amino groups on the PEI chains underwent Michael addition with the catechol/quinone groups on the PDA surface, achieving covalent grafting of PEI to obtain PDA@Cur-PEI nanoparticles (PCP). Amino-polyethylene glycol-carboxyl (NH2–PEG-COOH) was dissolved in distilled water, and the carboxyl group was activated with EDC·HCl and NHS for 2 h at room temperature. This mixture was then added dropwise to the PDA@Cur-PEI dispersion and reacted for another 2 h. The activated PEG-COOH underwent amide condensation with the amino groups on the PEI surface, forming stable amide bonds, yielding PDA@Cur–PEI-PEG (PCPP). For folic acid (FA) conjugation, FA, EDC·HCl, and NHS were dissolved in distilled water, activated for 2 h, then added to the nanoparticle dispersion and stirred for 2 h to obtain PDA@Cur–PEI-PEG-FA (PCPPF). Finally, the resulting PDA@Cur–PEI-PEG-FA nanoparticles were mixed with siRNA at different mass ratios (2:1 to 20:1) and allowed to stand at room temperature for 15–20 min. The positively charged PEI chains efficiently complexed the negatively charged siRNA via electrostatic interactions, forming stable nanocomplexes designated PDA@Cur–PEI-PEG-FA/siRela (PCPPFS).

2.3. Nanoparticle Characterization

Nanocomplexes with different PCPPF:siRNA ratios (w/w from 2 to 20) were loaded into a 2% agarose gel to evaluate the optimal ratio for complete siRNA loading using agarose gel electrophoresis. M5 Hipure Next III Gelred dye (Mei5bio, MF380–01) was added to the gel during preparation. The electrophoresis buffer was 1× TAE (50× TAE, Beyotime, ST716). Electrophoresis was performed at 150 V for 15 min. Zeta potential was measured using a Malvern Zetasizer to monitor the potential at different synthesis steps to confirm successful synthesis. The size and morphology of the nanoparticles were characterized using scanning electron microscopy (SEM) and transmission electron microscopy (TEM).

2.4. Cell Culture

The HK-2 cell line (SCSP-511) was purchased from the Cell Resource Center, Peking Union Medical College (Beijing, China). The TCMK-1 cell line (CCL-139) was obtained from the American Type Culture Collection (ATCC). All cell lines were authenticated by STR genotyping and confirmed to be mycoplasma-free. HK-2 cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin. TCMK-1 cells were cultured in DMEM supplemented with 10% FBS and 1% penicillin-streptomycin. Cells were maintained at 37 °C in a 5% CO2 humidified incubator. For subculturing, cells were washed with PBS, detached using 0.25% trypsin-EDTA at 37 °C for 3 min, neutralized with culture medium, centrifuged at 800g, and resuspended in fresh medium before seeding into new flasks.

2.5. Western Blotting

RIPA buffer supplemented with protease and phosphatase inhibitors was used to extract protein from cells and kidney tissues. After adding lysis buffer according to sample volume, the samples were vortexed and placed on ice for 40 min. After centrifugation at 1000 g for 5 min, the supernatant was collected, mixed with loading buffer, and stored at −80 °C. SDS-PAGE gels (10% or 12%) were prepared for protein electrophoresis. Electrophoresis was performed at 60 V for 50 min, followed by 120 V for 60 min. After electrophoresis, proteins were transferred onto preactivated PVDF membranes using the sandwich transfer method (250 mA constant current). The membranes were then blocked with 5% nonfat milk for 1 h at room temperature. After washing three times, the membranes were incubated with primary antibodies overnight at 4 °C on a shaker. After three washes, the membranes were incubated with secondary antibodies for 1 h at room temperature, washed three times, and visualized. β-actin was used as the loading control.

2.6. Real-Time Quantitative PCR (RT-qPCR)

Total RNA was extracted from cell pellets and kidney tissues using the FastPure Cell/Tissue Total RNA Isolation Kit V2 (Vazyme, RC112–01). RNA concentration was measured using a NanoDrop One spectrophotometer. cDNA was synthesized using the Hifair III first Strand cDNA Synthesis SuperMix for qPCR (Yeasen Biotec, 11141ES60). RT-qPCR was performed using Hieff UNICON Universal Blue qPCR SYBR Green Master Mix and primer solutions, following the manufacturer’s instructions. Primers were synthesized by Sangon Biotech, and sequences are listed in Table S2.

2.7. Cellular Uptake

Nanoparticle uptake was evaluated in LPS-stimulated HK-2 cells. HK-2 cells were seeded in confocal dishes and allowed to grow overnight. Cells were then treated with 10 μg/mL LPS for 24 h, followed by incubation with different nanoparticles (w/w 20, 50 nM siRNA; control nanoparticles received equivalent amounts without siRNA) for 6 h. PCPP and PCPPF were labeled with FITC. For dual labeling, FITC-labeled PCPP or PCPPF were complexed with Cy5.5-labeled siRNA to form PCPPS or PCPPFS, respectively. After incubation, cells were gently washed three times with PBS and fixed with 4% paraformaldehyde for 20 min at room temperature. After three washes, an antifade mounting medium containing DAPI (Lanboride, D0125) was added. Fluorescence images were acquired using a Leica TCS-SP8 confocal microscope.

2.8. Immunofluorescence

HK-2 cells seeded in confocal dishes were grown overnight. The treatment groups were pretreated with PCPPF or PCPPFS (w/w 20, 50 nM siRNA) for 24 h, then stimulated with 10 μg/mL LPS for 24 h. Cells were washed with PBS, fixed with 4% paraformaldehyde for 20 min at room temperature, and permeabilized with 0.3% Triton X-100 for 5 min. After washing, cells were blocked with 3% BSA for 30 min at room temperature, then incubated with primary antibodies overnight at 4 °C. After washing, cells were incubated with secondary antibodies for 1 h at 37 °C. Following washing, antifade mounting medium containing DAPI was added. Images were acquired using a Leica TCS-SP8 confocal microscope.

2.9. In Vitro Cytotoxicity

Cytotoxicity of PCPPFS was assessed using the CCK-8 kit (Lanboride, CK001). HK-2 cells seeded in 96-well plates were grown overnight. The treatment groups were pretreated with PCPPF or PCPPFS (w/w 20, 50 nM siRNA) for 24 h, then stimulated with 10 μg/mL LPS for 24 h. CCK-8 reagent was added according to the manufacturer’s instructions, and cell viability was measured using a microplate reader.

2.10. ROS Detection

HK-2 cells seeded in confocal dishes were grown overnight. Treatment groups were pretreated with PCPPF or PCPPFS (w/w 20, 50 nM siRNA) for 24 h, then stimulated with 10 μg/mL LPS for 24 h. Cells were then incubated with DCFH-DA probe and Hoechst 33342 staining solution for 20 min at 37 °C. After gentle washing with PBS, PBS was added, and images were quickly acquired using a Leica TCS-SP8 confocal microscope.

2.11. Animal Experiments

All animal experiments were approved by the Animal Ethics Committee of Peking University Third Hospital (Ethics Project No. A20250219). Eight-week-old male C57BL/6J mice were purchased from Vital River Laboratories. Mice were anesthetized with isoflurane inhalation. A 1 cm longitudinal incision was made in the midline of the abdomen, and the cecum was exposed. The cecum was ligated with a 4–0 silk suture at a location between the third and fourth vascular arches, then punctured twice through-and-through at the distal end of the ligation using a 25-gauge sterile needle, and a small amount of feces (∼1 mm in length) was gently extruded. The cecum was returned to the abdominal cavity, and the incision was closed in layers. All mice received subcutaneous injection of 1 mL sterile saline for fluid resuscitation and were placed on a heating pad until they recovered from anesthesia. The sham-operated control group underwent the same procedure without cecal ligation and puncture. Mice were euthanized 24 h after CLP surgery, and samples were collected for subsequent experiments.

2.12. In Vivo Therapeutic Evaluation

The therapeutic effect of PCPPFS was evaluated in CLP mice. Mice were randomly assigned to 4 groups: Group 1 (sham), Group 2 (untreated CLP), Group 3 (CLP + PCPPF), and Group 4 (CLP + PCPPFS). PCPPF and PCPPFS (0.5 mg/kg) were administered via tail vein injection 24 h before CLP surgery and immediately after surgery. At the end of the experiment, mice were anesthetized with isoflurane, and blood was collected from the retro-orbital venous plexus. Serum was separated for biochemical assays. Kidneys and other major organs were collected. Rela knockdown efficiency was assessed by RT-qPCR, Western blot, and immunohistochemistry. Changes in inflammatory markers, kidney injury markers, apoptotic pathway, and pyroptotic pathway after treatment were also evaluated.

2.13. Histological Staining (HE, PAS, TUNEL) and Immunohistochemistry

Mouse tissues were fixed in 4% paraformaldehyde at 4 °C for 18–24 h. After fixation, tissues were washed with PBS, then dehydrated through graded ethanol and precleared. Tissues were immersed in a melted mixture of paraffin and xylene for 30 min, then in pure paraffin (I) and (II) for 1 h each at 60 °C. Tissues were embedded in paraffin blocks, sectioned at 4–5 μm thickness, deparaffinized, and rehydrated before staining.

For HE staining, sections were stained with Harris hematoxylin, differentiated in 0.5% hydrochloric acid ethanol until nuclei appeared purple-red, blued under running tap water, then stained with 1% eosin. After dehydration and clearing, sections were mounted with neutral balsam. For PAS staining, sections were stained with 0.5–1% periodic acid solution, then with Schiff reagent for 30 min in the dark, washed, counterstained with hematoxylin, differentiated, blued, dehydrated, cleared, and mounted. For TUNEL staining, sections were deparaffinized, rehydrated, and incubated with Proteinase K. After equilibration, TdT reaction mixture was applied for 60 min at 37 °C in the dark. The reaction was terminated, and nuclei were stained with DAPI. For immunohistochemistry, sections underwent antigen retrieval, blocking of endogenous peroxidase, blocking with serum, incubation with primary antibody overnight at 4 °C, incubation with biotinylated secondary antibody, DAB development, counterstaining with hematoxylin, dehydration, clearing, and mounting.

2.14. Statistical Analysis

All data were analyzed using GraphPad Prism 10 software. Results are presented as mean ± standard error. Statistical parameters and the number of independent replicates are indicated in the figure legends. Two-tailed unpaired t test or Mann–Whitney U test was used for statistical analysis. P < 0.05, P < 0.01, or P < 0.001 was considered statistically significant; P > 0.05 was considered not significant.

3. Results and Discussion

3.1. In Vitro and In Vivo Validation of Increased Rela Expression in Septic AKI

The NF-κB pathway plays a key regulatory role in the inflammatory response of sepsis and is involved in the pathological process of AKI. Several active drugs for treating septic AKI exert their effects by inhibiting this pathway. − Rela is the core subunit of NF-κB and an indispensable protein for the function of this pathway. To verify whether Rela is highly expressed in septic AKI, we first examined Rela mRNA and protein expression levels in LPS-stimulated renal tubular epithelial cells and CLP-induced AKI mouse models. Compared with the control group, LPS stimulation of HK-2 cells for 24 h significantly increased Rela mRNA levels by approximately 2-fold (P < 0.01, Figure a). As shown in Figure b, renal Rela mRNA levels in CLP mice were 1.5-fold higher than those in sham mice (P < 0.05). Western blot analysis confirmed higher Rela protein expression in the model groups of HK-2 cells, TCMK-1 cells, and mouse kidneys (Figure c–e). Confocal microscopy revealed that in control HK-2 and TCMK-1 cells, Rela (green) primarily localized in the cytoplasm; after LPS stimulation, Rela showed increased nuclear accumulation and overall fluorescence intensity (Figure f,g). These results confirm that both transcription and protein levels of Rela are upregulated in the in vitro LPS model and the in vivo CLP model of septic AKI, providing experimental evidence for subsequent therapeutic strategies targeting Rela.

2.

2

Open in a new tab

Rela levels are elevated in in vitro and in vivo models. (a) Relative Rela mRNA levels in NC and LPS-treated HK-2 cells, n = 3. (b) Relative Rela mRNA levels in sham and CLP mice, n = 3. (c, d) Western blot of Rela expression in NC and LPS-treated HK-2 cells and TCMK-1 cells. (e) Western blot of Rela expression in sham and CLP mice. (f, g) Immunofluorescence confocal images of Rela in NC and LPS-treated HK-2 cells and TCMK-1 cells. Scale bar = 50 μm. (*P < 0.05, **P < 0.01, ***P < 0.001, ns = not significant).

3.2. Synthesis and Characterization of PCPPFS Nanoparticles

To achieve targeted combination therapy for septic AKI, we designed PCPPFS nanoparticles (Figure a). PDA served as the nanocarrier, adsorbing the therapeutic drug curcumin. Branched PEI was modified to bind siRNA. PEG was modified to improve solubility and biocompatibility. Finally, FA was modified to enable kidney targeting. The synthesis of PCPPFS was characterized using various methods. SEM and TEM images showed the morphology of the nanoparticles; PDA exhibited a spherical nanostructure, with slightly increased size after sequential modifications, ultimately resulting in PCPPFS nanoparticles with a diameter of approximately 110 nm (Figure c,d,f,g,h). Zeta potential was measured using a Malvern Zetasizer. As shown in Figure e, PDA had a zeta potential of approximately −23 mV. After curcumin loading, the potential shifted near zero. Following PEI modification, the nanoparticle potential became strongly positive (approximately +40 mV). After PEG and FA modifications, the potential remained positive, with PCPPF showing a zeta potential of approximately +19 mV, making it suitable as an siRNA carrier. A gel retardation assay showed partial siRNA retardation at a polymer/siRNA ratio of 2:1, and complete retardation at ratios of 4:1 and above (Figure i). Fourier-transform infrared (FTIR) spectroscopy was used to monitor the stepwise synthesis of PCPPF nanoparticles (Figure b). PDA displayed a broad absorption peak at 1612 cm–1, corresponding to overlapping quinone C  O and aromatic ring CC, and a peak at 1285 cm–1 for phenolic C–O stretching, confirming successful oxidative self-polymerization of dopamine. After curcumin loading, the PDA@Cur spectrum showed a sharpened peak at 1510 cm–1 and a new peak at 1036 cm–1 attributed to curcumin methoxyC–O–CH3 bending vibration, indicating successful curcumin loading. Following PEI coating, a peak at 1162 cm–1 appeared, characteristic of tertiary amine C–N stretching. The primary amine N–H bending (∼1635 cm–1) overlapped with the strong aromatic ring absorption of PDA/curcumin (1612 cm–1), resulting only in slight peak broadening. After EDC/NHS activation, the carboxyl group of NH2–PEG-COOH underwent amidation with surface amino groups of PEI. The FTIR spectrum showed complete disappearance of the free carboxyl acid peak at 1710 cm–1, appearance of a rounded broad peak at 1592 cm–1 (amide I band overlapping with carboxylate), and a characteristic peak at 842 cm–1 for PEG crystal −CH2– rocking vibration, confirming successful PEG grafting via covalent amide bonds. Finally, the γ-carboxyl group of FA was activated by EDC/NHS and reacted with residual amino groups of PEG via another amidation reaction. The spectrum displayed a strong sharp peak at 1604 cm–1 for pteridine/benzene ring CC stretching of folic acid, while the free carboxyl acid signal in the 1690–1720 cm–1 region disappeared, proving successful covalent modification of folic acid on the nanoparticle surface. This sequence of FTIR spectral evolution consistently indicates the successful construction of the multifunctional PDA@cur–PEI-PEG-FA nanoplatform.

3.

3

Open in a new tab

Synthesis and characterization of PCPPFS nanoparticles. (a) Schematic of PCPPFS nanoparticle synthesis. (b) FTIR spectra of PDA, PC, PCP, PCPP, and PCPPF. (c, d) SEM images of PDA and PCPPF. Scale bar = 100 nm. (e) Zeta potential of PDA, PC, PCP, PCPP, and PCPPF. (f–h) TEM images of PDA, PCPPF, and PCPPFS. Scale bar = 100 nm. (i) Gel retardation assay of PCPPFS.

3.3. In Vitro Cellular Uptake and Cytotoxicity of PCPPFS

Folic acid, a vitamin, is a classic targeting moiety for nanocarriers due to its high affinity for folate receptors. Many nanodelivery systems exploit high folate receptor expression at target sites for active targeting, such as in tumor therapy. − In AKI, several studies have used folic acid as a targeting group to develop kidney-targeting drugs or agents. − Because folate receptors are highly expressed on renal tubular epithelial cells, we hypothesized that the FA moiety on PCPPFS could enhance nanoparticle uptake in LPS-activated HK-2 cells. Under LPS treatment, uptake of FITC-labeled PCPPF was higher compared to FITC-labeled PCPP (Figure a). Next, we loaded Cy5.5-labeled siRela onto FITC-labeled nanoparticles. Under LPS treatment, HK-2 cells showed higher uptake of PCPPFS compared to PCPPS (Figure c). Furthermore, we assessed the cytotoxicity of the nanoparticles using the CCK-8 assay. As shown in Figure b, PCPPFS nanoparticles (w/w 20, 50 nM siRNA) did not significantly affect cell viability compared to the control group, indicating good safety at the cellular level. FA modification enhanced the uptake efficiency of PCPPFS nanoparticles in LPS-injured HK-2 cells, and the nanoparticles exhibited good cytocompatibility, laying the foundation for subsequent siRNA delivery and therapeutic effects.

4.

4

Open in a new tab

In vitro uptake, knockdown efficiency, and changes in NGAL and IL-6 upon PCPPFS treatment. (a) Uptake of PCPP and PCPPF by LPS-treated HK-2 cells. Scale bar = 50 μm. (b) Cell viability of untreated negative control and PCPPFS-treated (without LPS) HK-2 cells determined by CCK-8 assay, n = 3. (c) Uptake of PCPPS and PCPPFS by LPS-treated HK-2 cells. Scale bar = 50 μm. (d) Western blot of NGAL and RELA in different treatment groups. (e) Rela mRNA expression in different treatment groups, n = 3. (f) IL-6 mRNA expression in different treatment groups, n = 3. (g) Immunofluorescence images of RELA in different treatment groups. Scale bar = 50 μm. (*P < 0.05, **P < 0.01, ***P < 0.001, ns = not significant).

3.4. In Vitro Knockdown Efficiency of PCPPFS and Its Regulation on Inflammatory and Renal Injury Biomarkers

After verifying enhanced uptake, we evaluated whether PCPPFS could effectively silence Rela expression and reduce renal tubular cell injury and inflammation. RT-qPCR results (Figure e) showed that compared to the LPS group, PCPPF treatment reduced Rela mRNA levels by approximately 56% (P < 0.001), while PCPPFS treatment significantly reduced Rela mRNA levels by approximately 78% (P < 0.001). The knockdown efficiency of PCPPFS was significantly better than that of PCPPF (P < 0.05). Western blot results (Figure d) were consistent with the mRNA trends; the LPS group showed increased Rela protein expression, while PCPPFS treatment notably reduced band intensity. Immunofluorescence staining (Figure g) visually demonstrated the decrease in Rela green fluorescence and reduced nuclear accumulation after PCPPFS treatment. Meanwhile, after PCPPFS treatment, the protein level of the kidney injury marker NGAL (Figure d) decreased, and the reduction in IL-6 mRNA levels was approximately 74% (P < 0.001, Figure f); the PCPPFS group demonstrated an approximately 81% reduction compared to the LPS group (P < 0.001, Figure f). Both nanoparticles reduced intracellular NGAL expression and IL-6 mRNA levels, and the inhibitory effect of PCPPFS was stronger than that of PCPPF, indicating that the coloaded curcumin and therapeutic siRNA exert a synergistic effect, more effectively alleviating kidney injury and inflammation.

3.5. In Vitro Antiapoptotic, Antipyroptotic, and Oxidative Stress-Alleviating Effects of PCPPFS

Cell death modalities such as apoptosis and pyroptosis also contribute to the exacerbation of acute kidney injury. , Activation of the NF-κB pathway promotes the formation of the NLRP3 inflammasome, further facilitating pyroptosis. Additionally, NF-κB pathway activation leads to excessive inflammation, which may subsequently induce apoptosis. Oxidative stress is a key mechanism in the development and progression of AKI and chronic kidney disease, with ROS accumulation being a crucial factor in oxidative stress. Antioxidant therapy is a strategy for AKI treatment. Therefore, we further examined the effects of PCPPFS on the aforementioned pathways. We assessed changes in apoptosis and pyroptosis levels after nanoparticle treatment. Western blot results (Figure b) showed that compared to the LPS group, PCPPFS treatment significantly reduced the band intensity of the pro-apoptotic protein Bax and increased the band intensity of the antiapoptotic protein Bcl-2. In the pyroptosis pathway, the band intensities of NLRP3, cleaved Caspase-1, and GSDMD-N were all decreased. Immunofluorescence results (Figure a) also confirmed that both nanoparticles downregulated the expression of the pro-apoptotic protein Bax (green), with the PCPPFS treatment group showing weaker fluorescence intensity. ROS levels were detected using the DCFH-DA probe (Figure c). The LPS group exhibited intense green fluorescence in cells, indicating substantial ROS production. PCPPF and PCPPFS treatments significantly reduced the green fluorescence, suggesting effective alleviation of oxidative stress. Moreover, the inhibitory effect of PCPPFS was stronger than that of PCPPF, indicating that curcumin and siRela synergistically alleviate oxidative stress. CCK-8 results (Figure d) showed that cell viability in the LPS group dropped to approximately 44%; it recovered to approximately 82% in the PCPPF group (P < 0.001) and further to approximately 94% in the PCPPFS group (P < 0.001 vs LPS group; P < 0.05 vs PCPPF group). Therefore, PCPPFS exerts antiapoptotic, antipyroptotic, and oxidative stress-alleviating effects in vitro and improves cell viability. Rela knockdown inhibits NF-κB pathway activation, thereby downregulating apoptosis-related proteins (Bax/Bcl-2) and pyroptosis-related proteins (NLRP3/Caspase1/GSDMD); concurrently, curcumin nanoparticles scavenge ROS and suppress inflammation. The synergistic action of siRela and curcumin collectively alleviates LPS-induced HK-2 cell injury and improves cell survival, providing in vitro experimental support for in vivo treatment of septic AKI.

5.

5

Open in a new tab

In vitro antiapoptotic, antipyroptotic, and oxidative stress-alleviating effects of PCPPFS. (a) Immunofluorescence of Bax in different treatment groups. Scale bar = 50 μm. (b) Western blot of Bcl-2, Bax, cleaved GSDMD, NLRP3, and cleaved Caspase1 in different treatment groups. (c) ROS fluorescence levels in different treatment groups. Scale bar = 50 μm. (d) Cell viability in different treatment groups, n = 3. (*P < 0.05, **P < 0.01, ***P < 0.001, ns = not significant).

3.6. PCPPFS In Vivo Silencing and Amelioration of Renal Pathology and Inflammatory Biomarkers

To verify whether folic acid modification enhances targeted accumulation in AKI kidneys, we compared the distribution of PCPPS (without FA) and PCPPFS (with FA) in the kidneys of CLP mice. 24 h after intravenous injection, ex vivo kidney fluorescence imaging (Figure f) showed significantly higher fluorescence intensity in the PCPPFS group compared to the PCPPS group. This suggests that folic acid modification effectively enhances the accumulation of nanoparticles in injured kidneys.

6.

6

Open in a new tab

In vivo knockdown efficiency of PCPPFS and changes in renal histopathology, NGAL, and IL-6 in mice. (a) HE and PAS staining of different treatment groups. Scale bar = 20 μm. (b) Western blot of NGAL and RELA in different treatment groups. (c, d) Rela and IL-6 mRNA expression in different treatment groups, n = 3. (*P < 0.05, **P < 0.01, ***P < 0.001, ns = not significant). (e) Immunohistochemistry of NGAL and RELA in different treatment groups. Scale bar = 50 μm. (f) Kidney uptake of PCPPS and PCPPFS in l mice. Scale bar = 50 μm.

After tail vein injection of nanoparticles, we evaluated renal histopathology in different treatment groups. As shown in Figure a, compared to the CLP group, PCPPF and PCPPFS treatments resulted in more intact tubular structures and clearer brush borders; the improvement was more pronounced in the PCPPFS group. We also detected renal Rela levels in different treatment groups using immunohistochemistry, Western blot, and RT-qPCR (Figure b,c,e). Compared to CLP mice and the PCPPF-treated group, the PCPPFS-treated group showed reduced renal Rela levels at both mRNA and protein levels. Specifically, RT-qPCR results (Figure c) showed that compared to the CLP group, the PCPPFS-treated group exhibited a significant reduction in renal Rela mRNA levels by approximately 73% (P < 0.001), while the PCPPF group showed a reduction of approximately 41% (P < 0.001). The knockdown efficiency of PCPPFS was significantly superior to that of PCPPF (P < 0.001). IL-6 mRNA levels (Figure d) were reduced by over 90% in the PCPPFS group (P < 0.001). Simultaneously, PCPPFS reduced NGAL expression and IL-6 mRNA levels in kidney tissues (Figure b, d, e), suggesting it may also exert anti-inflammatory effects. Immunohistochemical staining (Figure e) showed dense NGAL-positive areas (brown) in the kidneys of CLP mice, which were significantly reduced after PCPPFS treatment. These results indicate that PCPPFS nanoparticles effectively target the kidneys, significantly knock down Rela expression, inhibit renal NGAL expression and IL-6 transcription, and ameliorate renal histopathological damage in septic AKI mice, demonstrating good therapeutic potential in vivo. In addition, the hemocompatibility and hepatorenal toxicity of this nanoformulation were preliminarily evaluated (Figures S1 and S2).

3.7. PCPPFS Inhibits Apoptosis and Pyroptosis in Kidney Tissues of CLP Mice and Improves Renal Function

We further evaluated the effects of PCPPFS on pyroptosis and apoptosis pathways in kidney tissues of CLP mice. As shown in Figure a, TUNEL staining of mouse kidney tissues revealed green signals indicating apoptotic nuclei. The CLP group showed numerous apoptotic cells in the kidney, which decreased in the PCPPF group and were only sporadically observed in the PCPPFS group. Western blot assessed the protein levels of Bcl-2, Bax, cleaved GSDMD, NLRP3, and cleaved Caspase1 in different treatment groups (Figure b). The Western blot results from mouse kidney tissues were consistent with the in vitro findings; PCPPFS treatment inhibited the expression of Bax, NLRP3, cleaved Caspase-1, and GSDMD-N, while upregulating Bcl-2. Immunohistochemistry for Bax (Figure a) similarly supported this conclusion. Additionally, we evaluated the effect of the nanoparticles on renal function in CLP mice by measuring serum levels of creatinine (Crea), urea, and cystatin C (CysC). Both nanoparticle treatments improved renal function parameters in CLP mice. Compared to the CLP group, the PCPPFS group showed significantly decreased serum Crea, urea, and CysC levels. Specifically, relative to the CLP group, Crea levels (Figure c) were reduced by approximately 61% in the PCPPFS group (P < 0.01), urea levels (Figure d) by approximately 86% (P < 0.001), and CysC levels (Figure e) by approximately 69% (P < 0.001). These results indicate that PCPPFS effectively inhibits apoptosis and pyroptosis of renal tubular epithelial cells in septic AKI mice and significantly improves renal function.

7.

7

Open in a new tab

PCPPFS inhibits apoptosis and pyroptosis in kidney tissues of CLP mice and improves renal function. (a) Immunohistochemistry of BAX and TUNEL staining in different treatment groups. Scale bars = 50 and 100 μm, respectively. (b) Western blot of Bcl-2, Bax, cleaved GSDMD, NLRP3, and cleaved Caspase1 in different treatment groups. (c–e) Serum levels of Crea, urea, and CysC in different treatment groups, n = 5. (*P < 0.05, **P < 0.01, ***P < 0.001, ns = not significant).

4. Conclusion

In summary, we synthesized a targeted bifunctional siRNA nanoparticle that effectively knocked down Rela at both the cellular level and in mouse kidney tissues. Together with the loaded curcumin, it synergistically treats septic acute kidney injury. At both in vitro and in vivo levels, PCPPFS exerted antiapoptotic, antipyroptotic, and inflammation-reducing (IL-6) effects, and reduced the kidney injury molecule NGAL. In CLP mice, PCPPFS improved renal function and renal pathology, exhibited low hepatorenal toxicity in normal mice, and demonstrated good biosafety, making it a promising therapeutic agent for septic AKI.

Supplementary Material

ao6c07405_si_001.pdf (198.3KB, pdf)

The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acsomega.6c07405.

  • Hemocompatibility and hepatorenal toxicity of this nanoformulation (Figures S1–S2); siRNA sequences and RT-qPCR primer sequences (Tables S1–S2) (PDF)

X.X., S.Y., and L.C. designed the experimental plan. X.X., J.T., Z.S., Y.N., and H.G. conducted the cell experiments. X.X., J.T., Q.Z., H.W., and C.W. carried out the animal experiments. X.X., Q.L., Z.L., D.L., J.P., and C.M. wrote the paper. L.C. improved and supervised the submitted and revised manuscript. All authors read and approved the final manuscript.

This work is supported by the Special Project for Health Development Research of the Capital and the Mindray Joint Project (2026-M3–2–409–1).

The study was approved by the Ethics Committee of Peking University Third Hospital (Approval No.: A20250219).

The authors declare no competing financial interest.

References

  1. Pais T., Jorge S., Lopes J. A.. Acute Kidney Injury in Sepsis. Int. J. Mol. Sci. 2024;25(11):5924. doi: 10.3390/ijms25115924. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ostermann M., Lumlertgul N., Jeong R., See E., Joannidis M., James M.. Acute kidney injury. Lancet. 2025;405(10474):241–256. doi: 10.1016/S0140-6736(24)02385-7. [DOI] [PubMed] [Google Scholar]
  3. Kounatidis D., Vallianou N. G., Psallida S., Panagopoulos F., Margellou E., Tsilingiris D., Karampela I., Stratigou T., Dalamaga M.. Sepsis-Associated Acute Kidney Injury: Where Are We Now? Medicina. 2024;60(3):434. doi: 10.3390/medicina60030434. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Yubolphan R., Kobroob A., Kongkaew A., Chiranthanut N., Jinadang N., Wongmekiat O.. Berberine Mitigates Sepsis-Associated Acute Kidney Injury in Aged Rats by Preserving Mitochondrial Integrity and Inhibiting TLR4/NF-κB and NLRP3 Inflammasome Activations. Antioxidants. 2024;13(11):1398. doi: 10.3390/antiox13111398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ji Y., Zhao Z., Yang Y., Wang X., Qiao R., Yu X., Gong X., Feng Z., Hong Q.. Mechanisms Underlying the Impact of Interleukin Family on Acute Kidney Injury: Pathogenesis, Progression, and Therapy. Research. 2025;8:0738. doi: 10.34133/research.0738. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ren Q., Guo F., Tao S., Huang R., Ma L., Fu P.. Flavonoid fisetin alleviates kidney inflammation and apoptosis via inhibiting Src-mediated NF-κB p65 and MAPK signaling pathways in septic AKI mice. Biomed. Pharmacother. 2020;122:109772. doi: 10.1016/j.biopha.2019.109772. [DOI] [PubMed] [Google Scholar]
  7. Fu H., Hu Z., Di X., Zhang Q., Zhou R., Du H.. Tenuigenin exhibits protective effects against LPS-induced acute kidney injury via inhibiting TLR4/NF-κB signaling pathway. Eur. J. Pharmacol. 2016;791:229–234. doi: 10.1016/j.ejphar.2016.08.013. [DOI] [PubMed] [Google Scholar]
  8. Tang Y., Wang C., Chen S., Li L., Zhong X., Zhang J., Feng Y., Wang L., Chen J., Yu M., Wang F., Wang L., Li G., He Y., Li Y.. Dimethyl fumarate attenuates LPS induced septic acute kidney injury by suppression of NFκB p65 phosphorylation and macrophage activation. Int. Immunopharmacol. 2022;102:108395. doi: 10.1016/j.intimp.2021.108395. [DOI] [PubMed] [Google Scholar]
  9. Li T., Sun H., Li Y., Su L., Jiang J., Liu Y., Jiang N., Huang R., Zhang J., Peng Z.. Downregulation of macrophage migration inhibitory factor attenuates NLRP3 inflammasome mediated pyroptosis in sepsis-induced AKI. Cell Death Discovery. 2022;8(1):61. doi: 10.1038/s41420-022-00859-z. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Yang Y., Xu J., Tu J., Sun Y., Zhang C., Qiu Z., Xiao H.. Polygonum cuspidatum Sieb. et Zucc. Extracts improve sepsis-associated acute kidney injury by inhibiting NF-?B-mediated inflammation and pyroptosis. J. Ethnopharmacol. 2024;319:117101. doi: 10.1016/j.jep.2023.117101. [DOI] [PubMed] [Google Scholar]
  11. Cao Y., Liu X., Guo C., Yang W., Wang X., Wang X., Xu H., Wang W., Liu D., Zhang J., Cui W., Chen Y., Guo X., Chen D.. Biomimetic reactive oxygen/nitrogen nanoscavengers inhibit ″ferroptosis storm″ and modulate immune targeting for acute kidney injury. J. Controlled Release. 2025;379:59–76. doi: 10.1016/j.jconrel.2025.01.006. [DOI] [PubMed] [Google Scholar]
  12. Huang X., Zhang F., Yang Y., Liu J., Tan X., Zhou P., Tang X., Hu J., Chen L., Yuan M., Zheng G., Xu Z., Qiu Z.. Curcumin-copper complex nanoparticles as antioxidant nanozymes for acute kidney injury alleviation. Mater. Today Bio. 2025;32:101794. doi: 10.1016/j.mtbio.2025.101794. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Cai Y., Huang C., Zhou M., Xu S., Xie Y., Gao S., Yang Y., Deng Z., Zhang L., Shu J., Yan T., Wan C. C.. Role of curcumin in the treatment of acute kidney injury: research challenges and opportunities. Phytomedicine. 2022;104:154306. doi: 10.1016/j.phymed.2022.154306. [DOI] [PubMed] [Google Scholar]
  14. Huang J., Guo J., Dong Y., Xiao H., Yang P., Liu Y., Liu S., Cheng S., Song J., Su Y., Wang S.. Self-assembled hyaluronic acid-coated nanocomplexes for targeted delivery of curcumin alleviate acute kidney injury. Int. J. Biol. Macromol. 2023;226:1192–1202. doi: 10.1016/j.ijbiomac.2022.11.233. [DOI] [PubMed] [Google Scholar]
  15. Li H., Sun H., Xu Y., Xing G., Wang X.. Curcumin plays a protective role against septic acute kidney injury by regulating the TLR9 signaling pathway. Transl. Androl. Urol. 2021;10(5):2103–2112. doi: 10.21037/tau-21-385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Yang Y., Xiao Z., Liu L., Hu T., Liu Y., Wang D.. Curcumin regulates ferroptosis by activating α7 nicotinic acetylcholine receptor to improve sepsis-induced acute kidney injury. Renal Failure. 2026;48(1):2657652. doi: 10.1080/0886022X.2026.2657652. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Luo M., Zhu Q., Xu G., Liu D., Xiao J., Shi Q.. Intervention effect of curcumin on sepsis-associated acute kidney injury via regulation of p300 expression and protein lactylation. BMC Immunol. 2025;26(1):67. doi: 10.1186/s12865-025-00750-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Wang L., Wu N., Zhang W., Huang D., Li Y.. Curcumin Alleviates Sepsis-Associated Acute Kidney Injury Potentially by Inhibiting Ferroptosis Through the ACSL4/GPX4 Signaling Pathway. Drug Dev. Res. 2025;86(7):e70181. doi: 10.1002/ddr.70181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lan T. Y., Guo H. L., Lu X., Geng K. D., Wu L., Luo Y. J., Zhu J. F., Shen X. C., Guo Q. Q., Wu S. Z.. Dual-Responsive Curcumin-Loaded Nanoparticles for the Treatment of Cisplatin-Induced Acute Kidney Injury. Biomacromolecules. 2022;23(12):5253–5266. doi: 10.1021/acs.biomac.2c01083. [DOI] [PubMed] [Google Scholar]
  20. Guo H., Lan T., Lu X., Geng K., Shen X., Mao H., Guo Q.. ROS-responsive curcumin-encapsulated nanoparticles for AKI therapy via promoting lipid degradation in renal tubules. J. Mater. Chem. B. 2024;12(12):3063–3078. doi: 10.1039/D3TB02318D. [DOI] [PubMed] [Google Scholar]
  21. Chen X., Ren Q., Chen G., Yi Z., Tong Q., Ran Y., Ma L., Fu P., Ma L., Li X.. Self-Assembled Phytochemical Nanomedicines with Enhanced Bioactivities for Effective Acute Kidney Injury Therapy. ACS Sustainable Chem. Eng. 2023;11(19):7288–7300. doi: 10.1021/acssuschemeng.2c05786. [DOI] [Google Scholar]
  22. Xie X., Yue T., Gu W., Cheng W., He L., Ren W., Li F., Piao J.-G.. Recent Advances in Mesoporous Silica Nanoparticles Delivering siRNA for Cancer Treatment. Pharmaceutics. 2023;15(10):2483. doi: 10.3390/pharmaceutics15102483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sargazi S., Arshad R., Ghamari R., Rahdar A., Bakhshi A., Karkan S. F., Ajalli N., Bilal M., Diez-Pascual A. M.. siRNA-based nanotherapeutics as emerging modalities for immune-mediated diseases: A preliminary review. Cell Biol. Int. 2022;46(9):1320–1344. doi: 10.1002/cbin.11841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Friedrich M., Aigner A.. Therapeutic siRNA: State-of-the-Art and Future Perspectives. Biodrugs. 2022;36(5):549–571. doi: 10.1007/s40259-022-00549-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Chen Q., Yang Y., Ying X., Huang C., Chen J., Wang J., Wu Z., Zeng W., Miao C., Shi X., Nan Y., Huang Q., Ai K.. Hierarchical Targeting Nanodrug with Holistic DNA Protection for Effective Treatment of Acute Kidney Injury. Adv. Sci. 2025;12(6):2411254. doi: 10.1002/advs.202411254. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Liang Z., He Y., Ieong C. S. U., Choi C. H. J.. Cell-nano interactions of polydopamine nanoparticles. Curr. Opin. Biotechnol. 2023;84:103013. doi: 10.1016/j.copbio.2023.103013. [DOI] [PubMed] [Google Scholar]
  27. Chen L., Chen S., Yi Z., Wu X., Zhong S., Mao L., Wang Z., Shuai Q., Li X.. Oligosaccharide and alkyl chain-modified polyethyleneimines for efficient siRNA delivery. Eur. Polym. J. 2025;233:113988. doi: 10.1016/j.eurpolymj.2025.113988. [DOI] [Google Scholar]
  28. Guan X., Wang L.-l., Yang Z., Wang Y., Zhao Y., Wang D.. pH-responsive 2D niobium carbide nanosheets for targeted circPUM1 siRNA delivery in ovarian cancer therapy. Mater. Today Bio. 2025;35:102314. doi: 10.1016/j.mtbio.2025.102314. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Chunling W., Xiaoqian W., Wei D., Ningxin Z., Chen G., Yanlu X., Zhuo S., Xiaorui W., Minghao G., Ruizhe Z., Yan X., Liting W.. Tectorigenin could alleviate SA-AKI via reducing M1-like polarization of macrophages through KLF4/NF-κB pathway. Int. Immunopharmacol. 2025;164:115300. doi: 10.1016/j.intimp.2025.115300. [DOI] [PubMed] [Google Scholar]
  30. Liu T., Zhang Y., Liu X., Zuo Z.. Fortunellin attenuates sepsis-induced acute kidney injury by inhibiting inflammation and ferroptosis via the TLR4/NF-κB pathway. Histol. Histopathol. 2025;40(7):1083–1093. doi: 10.14670/HH-18-841. [DOI] [PubMed] [Google Scholar]
  31. Lou Y., Shi H., Sha N., Li F., Gu X., Lin H.. Ursodeoxycholic acid protects against sepsis-induced acute kidney injury by activating Nrf2/HO-1 and inhibiting NF-κB pathway. BMC Nephrol. 2025;26(1):45. doi: 10.1186/s12882-025-03977-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Shou D.-W., Li Y.-R., Xu X.-J., Dai M.-H., Zhang W., Yang X., Tu Y.-X.. Parthenolide Attenuates Sepsis-Induced Acute Kidney Injury in Rats by Reducing Inflammation. Evidence-Based Complementary Altern. Med. 2023;2023:8759766. doi: 10.1155/2023/8759766. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Han T., Jiang Y., Ge W., Lu Y., Liu R., Sun Z.. 2,5-Dihydroxyacetophenone attenuates acute kidney injury induced by intra-abdominal infection in rats. Nephrology. 2024;29(10):636–644. doi: 10.1111/nep.14335. [DOI] [PubMed] [Google Scholar]
  34. Shou D.-W., Yu Z.-L., Meng J.-B., Lai Z.-Z., Pang L.-S., Dai M.-H., Yang X., Tu Y.-X.. Panax notoginseng Alleviates Sepsis-Induced Acute Kidney Injury by Reducing Inflammation in Rats. Evidence-Based Complementary Altern. Med. 2022;2022:9742169. doi: 10.1155/2022/9742169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wang L., Evans J. C. C., Ahmed L., Allen C.. Folate receptor targeted nanoparticles containing niraparib and doxorubicin as a potential candidate for the treatment of high grade serous ovarian cancer. Sci. Rep. 2023;13(1):3226. doi: 10.1038/s41598-023-28424-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Werner M. E., Karve S., Sukumar R., Cummings N. D., Copp J. A., Chen R. C., Zhang T., Wang A. Z.. Folate-targeted nanoparticle delivery of chemo- and radiotherapeutics for the treatment of ovarian cancer peritoneal metastasis. Biomaterials. 2011;32(33):8548–8554. doi: 10.1016/j.biomaterials.2011.07.067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Ibrahim A. I., Sahyon H. A., Attia A. M., El-Shehawy A. A.. Novel protocatechuic acid encapsulated bovine serum albumin functionalized folic acid nanoparticles for targeted therapy in urethane-induced lung cancer model. Sci. Rep. 2025;15(1):25793. doi: 10.1038/s41598-025-08465-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Knight S. F., Kundu K., Joseph G., Dikalov S., Weiss D., Murthy N., Taylor W. R.. Folate Receptor-Targeted Antioxidant Therapy Ameliorates Renal Ischemia-Reperfusion Injury. J. Am. Soc. Nephrol. 2012;23(5):793–800. doi: 10.1681/ASN.2011070711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Li X., Cantley L. G., Xu L.. Engineering polyamidoamine dendrimer nanoparticles for targeted drug delivery to proximal tubular cells after renal ischemia/reperfusion injury. J. Controlled Release. 2025;388:114323. doi: 10.1016/j.jconrel.2025.114323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Du B., Zhao M., Wang Y., Yu L., Jiao Q., Bai Y., Cheng G.. Folic acid-targeted pluronic F127 micelles improve oxidative stress and inhibit fibrosis for increasing AKI efficacy. Eur. J. Pharmacol. 2022;930:175131. doi: 10.1016/j.ejphar.2022.175131. [DOI] [PubMed] [Google Scholar]
  41. Huang C., Zeng T., Li J., Tan L., Deng X., Pan Y., Chen Q., Li A., hu J.. Folate Receptor-Mediated Renal-Targeting Nanoplatform for the Specific Delivery of Triptolide to Treat Renal Ischemia/Reperfusion Injury. ACS Biomater. Sci. Eng. 2019;5(6):2877–2886. doi: 10.1021/acsbiomaterials.9b00119. [DOI] [PubMed] [Google Scholar]
  42. Xia W., Li Y., Wu M., Jin Q., Wang Q., Li S., Huang S., Zhang A., Zhang Y., Jia Z.. Gasdermin E deficiency attenuates acute kidney injury by inhibiting pyroptosis and inflammation. Cell Death Dis. 2021;12(2):139. doi: 10.1038/s41419-021-03431-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. He J., Chen Y., Li Y., Feng Y.. Molecular mechanisms and therapeutic interventions in acute kidney injury: a literature review. BMC Nephrol. 2025;26(1):144. doi: 10.1186/s12882-025-04077-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Fu Y., Cao J., Wei X., Ge Y., Su Z., Yu D.. Klotho alleviates contrast-induced acute kidney injury by suppressing oxidative stress, inflammation, and NF-KappaB/NLRP3-mediated pyroptosis. Int. Immunopharmacol. 2023;118:110105. doi: 10.1016/j.intimp.2023.110105. [DOI] [PubMed] [Google Scholar]
  45. Yin H., Yan Q., Li Y., Tang H.. Dihydromyricetin Nanoparticles Alleviate Lipopolysaccharide-Induced Acute Kidney Injury by Decreasing Inflammation and Cell Apoptosis via the TLR4/NF-κB Pathway. J. Funct. Biomater. 2024;15(9):249. doi: 10.3390/jfb15090249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Li Y., Wang J., Huang D., Yu C.. Baicalin Alleviates Contrast-Induced Acute Kidney Injury Through ROS/NLRP3/Caspase-1/GSDMD Pathway-Mediated Proptosis in vitro. Drug Des. Dev. Ther. 2022;16:3353–3364. doi: 10.2147/DDDT.S379629. [DOI] [PMC free article] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

ao6c07405_si_001.pdf (198.3KB, pdf)

Articles from ACS Omega are provided here courtesy of American Chemical Society

RESOURCES