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. 2024 May 2;9(19):21144–21151. doi: 10.1021/acsomega.4c01025

Green Synthesis of Gold Nanoparticles from Polygahatous Polysaccharides and Their Anticancer Effect on Hepatic Carcinoma through Immunoregulation

Maodong Leng †,‡, Huiqin Jiang §, Sitong Zhang †, Yixi Bao †,*
PMCID: PMC11097183  PMID: 38764635

Abstract

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Hepatic carcinoma is one of the leading causes of morbidity and mortality among all cancers, but no effective treatment measures have been developed. Herein, polystyrene polysaccharide (PSP) extracted from Polygonatum was used to synthesize gold nanoparticles (PSP-AuNPs) by heating and reduction methods, and the characteristics of the PSP-AuNPs were detected after successful synthesis. In vitro, the immunoregulatory effects of PSP-AuNPs were studied by testing the concentrations of NO, TNF-α, and IL-12p70 in the culture media of PSP-AuNPs-treated RAW264.7 macrophages, and the effect of biocompatibility on the viability of RAW264.7 macrophages and L02 cells was studied via a CCK-8 assay. In vivo, tumor-bearing mice were established and treated with PSP-AuNPs, and the anticancer effects were studied by detecting trends in tumor volume, tumor inhibition rate, and tumor cell proliferation index. Immunoregulation was assessed by evaluating the serum levels of TNF-α and IL-10, the CD4+/CD8+ lymphocyte ratio in peripheral blood and the spleen and thymus indices; toxicity was investigated by measuring body weight, liver and renal function indices. The results showed that PSP-AuNPs could regulate immune function both in vitro and in vivo with almost no toxicity. PSP-AuNPs exhibited excellent anticancer effects on hepatic carcinoma in vivo. The anticancer effect could be strengthened, and the toxicity could be reduced by the combined use of PSP-AuNPs and ADM. In conclusion, PSP-AuNPs could be effective as a therapy and adjuvant therapy for treating hepatic carcinoma, providing potential treatment strategies for this disease.

1. Introduction

Hepatic carcinoma is one of the most common cancers worldwide, with morbidity and mortality rates ranking sixth and third respectively among all cancers.1 The incidence of hepatic cancer remains stable among men, but obesity and hepatitis B and C virus infections have contributed to the stable increase in incidence among women (1.6–1.7% annually).2 Current therapeutic methods for treating hepatic carcinoma include surgery, targeted treatment, and immunotherapy, but these methods are restricted for use due to several limitations. Surgery is an effective treatment method for hepatic carcinoma, but it is not suitable for the majority of patients because of the usually advanced stages of hepatic carcinoma at diagnosis.3 Targeted treatment with sorafenib has achieved significant effect in the systemic treatment, but it can be ineffective due to the abnormal heterogeneity of hepatic carcinoma and rapid development of drug resistance, and the 3-month survival rate is still low.4−6 Immune checkpoint inhibitors such as nivolumab, altizolizumab, and bevacizumab are used for immunotherapy of hepatic carcinoma, and relatively good effects, including prolonged overall survival (OS) and progression-free survival (PFS), can be obtained.7−10 However, the effects of immunotherapy are still limited, and hepatic carcinoma is prone to recurrence and distant metastasis. In addition, targeted therapy and immunotherapy in hepatic carcinoma could cause various adverse effects such as gastrointestinal toxicity, hepatotoxicity, and endocrine-related toxicity. Therefore, there is an urgent need to develop new treatments for hepatic carcinoma.

Traditional Chinese medicine (TCM) can be used as an adjuvant therapy for radiation and chemotherapy to strengthen therapeutic effects and reduce adverse effects.11,12 Components, including polysaccharides, phenolic acids, flavonoids, esters, and cumarin, can be purified from TCMs via modern purification methods.13 Polysaccharides purified from TCMs can enhance immunity without causing toxicity during cancer therapy.14,15Polygonatum is the dry rhizoma of the plant Polygonatum sibiricum. Polygahatous polysaccharide (PSP) is extracted from Polygonatum and has various biological functions, including immunoregulation, antioxidant activity, reducing diabetes complications, preventing osteoporosis, and exerting antitumor effects.16−20 However, the immunoregulatory function and anticancer effect of PSP on hepatic carcinoma are unknown.

In recent years, an increasing number of nanomaterials have been studied for the treatment of hepatic carcinoma, and these materials can deliver small molecular substances such as chemotherapeutics, polysaccharides, and therapeutic agents; increase dispersion, solubility, and stability; and change pharmacokinetics.21−23 Uptake of the anticancer cargo inside the cells remains a major challenge in the application of nanomaterials; membrane potential and receptor-mediated uptake of nanoparticles are important factors, and receptor-mediated approach has been confirmed as a useful method to increase the uptake of nanomaterial drugs inside hepatic cancer cells.24,25 Gold nanoparticles (AuNPs) are the most widely studied type of nanoparticle, and they can be easily combined with drugs and increase the stability of drugs due to their optical, plasmon resonance, and biological conjugation properties.26 AuNPs possess advantages, including low toxicity, slowing drug degradation, and reducing the dosage and toxicity of drugs used.27 AuNPs have been widely used in biomedical fields, including clinical chemistry, immunology, microbiology, the diagnosis, and treatment of malignant tumors.28,29 Previous studies have demonstrated that the effects of the combination of AuNPs with extracts and polysaccharides from TCM are superior to the effects of single TCM ingredients in the treatment of liver cancer.30,31 The most common method for synthesizing AuNPs involves the reduction of dissolving gold salt, such as chloroauric acid. Therefore, AuNPs might be synthesized through the reduction of chloroauric acid with PSP (PSP-AuNPs), reducing the toxicity of drugs and extending the duration of drug circulation to achieve more efficient biological functions.

In this study, we will synthesize PSP-AuNPs via a green method and investigate the role of these nanoparticles in the treatment and adjuvant treatment of hepatic carcinoma through immunoregulation. In contrast to the earlier approaches for hepatic carcinoma treatments, this study will provide a new strategy for therapy. The new method for hepatic carcinoma therapy will increase the uptake of drugs in cells due to the use of AuNPs, reduce the adverse effects caused by targeted and immune inhibitor chemotherapy, and provide an alternative therapy method for patients at advanced stages who lose the opportunity of surgery.

2. Materials and Methods

2.1. Synthesis of PSP-AuNPs

The PSP powder used in this study was purchased from Sichuan Weikeqi Biological Technology Company (Chengdu, China) and had purity greater than 90% and was free of endotoxin. PSP-AuNPs were synthesized with the use of PSP and HAuCl4 (1 M) under heating conditions. To obtain the optimal conditions for the synthesis, we continuously changed the following reaction conditions: concentration of PSP (10 mg/mL-0.5 mg/mL); reaction volume (0.5 mL-5 mL); amount of HAuCl4 (3 μL, 2.5 μL, 2 μL, 1.5 μL, 1.25 μL, 1 μL, 0.5 μL, 0.2 μL); reaction time (10 min, 15 min, 30 min, 45 min); and reaction temperature (60 °C, 80 °C, 100 °C). When the color of the solution changed from light yellow to ruby red, the synthesis of PSP-AuNPs was preliminarily considered successful26 and further tests were performed to analyze the synthesis and characterization of the nanoparticles. In addition, sodium citrate dihydrate and HAuCl4 (1 M) were used to synthesize AuNPs as controls.32

2.2. Characterization of PSP-AuNPs

The ultraviolet–visible absorption spectrum (UV–VIS, range: 200–800 nm) was used to detect the PSP, PSP-AuNP, and AuNP solutions to confirm the success of the synthesis and analyze the absorption peak. Transmission electron microscopy (TEM) and atomic force microscopy (AFM) were used for the detection of the size and morphological characteristics (TEM for the overall shape and AFM for the surface morphology) of PSP-AuNPs and AuNPs, Fourier transform infrared spectroscopy (FTIR) was used to analyze the functional groups of PSP-AuNPs, dynamic light scattering (DLS) was used to detect the size and zeta distributions of PSP-AuNPs, and energy dispersive X-ray (EDX) was used to characterize the energy spectrum of PSP-AuNPs.

2.3. Cell Culture

RAW264.7 macrophages were donated by the Clinical Laboratory Department of the Second Affiliated Hospital of Chongqing Medical University. To prevent differentiation, RAW264.7 macrophages were cultured in specialized complete DMEM (Procell, Wuhan, China). We obtained H22 cells from Procell, and RPMI-1640 culture medium supplemented with 10% fetal bovine serum (FBS) was used for culture. RPMI-1640 supplemented with 20% FBS was used to culture L02 cells (Cellverse Bioscience, Shanghai, China). All the culture media were supplemented with 1% penicillin and streptomycin. The culture conditions for the three cell lines in the cell incubator were 5% CO2 and 37 °C.

2.4. Tests of NO, TNF-α, and IL-12p70 Levels

The in vitro immunoregulatory function of PSP-AuNPs was investigated in RAW264.7 cells. Equal amounts of RAW264.7 cells (5 × 104 cells/well) were seeded in each well of 24-well plates. After 24 h of adhesion and adaptation, the RAW264.7 cells were treated with LPS (100 ng/mL; ACMEC, China), different concentrations of PSP and PSP-AuNPs (100–500 μg/mL) separately. Cells not treated (normal control group, NC) and treated with LPS were regarded as negative and positive controls, respectively. The supernatants were collected and centrifuged for immunoregulation tests after 24 h of treatment. The Griess reaction was used to detect NO levels, and the levels of TNF-α and IL-12p70 were detected via ELISA. The tests were performed with a Micro Nitric Oxide (NO) Assay Kit (Abbkine, USA) and an ELISA Kit (Elabscience, Wuhan, China) according to the manufacturers’ instructions.

2.5. CCK-8 Assay

RAW264.7 macrophages and L02 cells were plated on 96-well plates with equal amounts of cells in each well (5 × 103 cells/well). After 24 h of incubation, both cell lines were treated with different concentrations of PSP and PSP-AuNPs (100–500 μg/mL). In addition, RAW264.7 macrophages and L02 cells were treated with LPS (100 ng/mL) and adriamycin (ADM) (2 μg/mL; HARVEYBIO, China), respectively, as controls. Normal controls without any treatment (NC) were used for both cell lines. After 24 h of treatment, 10 μL of CCK-8 solution (Biosharp Life Sciences, China) was added to each well. The plates were incubated for more than 1.5 h to react before the absorbance was measured at 450 nm with an ELISA instrument. The following formula was used for the calculation of cell viability: cell viability (%) = [OD (experiment) – OD (blank)]/[OD (NC) – OD (blank)] × 100%.

2.6. Animals

We purchased female C57BL/6 mice from the Laboratory Animal Center of Zhengzhou University. The weights of the mice were 16–18 g, and the mice were approximately 6 weeks old. Hepatic carcinoma tumor-bearing mice model was established through injecting H22 cells (4 × 105) into the subcutaneous right axilla of the mice. The tumor-bearing mice were randomly assigned to one of the following groups: (1) normal saline (NS) (400 μL/mouse/day), (2) PSP (200 mg/kg/day), (3) PSP-AuNPs (200 mg/kg/day), (4) ADM (4 mg/kg/3 days), and (5) PSP-AuNPs + ADM. Five mice were included in each group, and the drugs were administered via intraperitoneal injection.

During administration, the body weights of the mice and the diameters of the tumors were measured with a laboratory balance and a Vernier caliper every 3 days. The longest (a) and shortest (b) diameters of the tumors were obtained, and a × b2/2 was used to calculate the tumor volume.33 Moreover, the activities and mental states of the mice were recorded. The mice were administered with these agents for 21 days, then the mice were sacrificed by cervical dislocation. Peripheral blood was collected from the mice into heparin lithium-anticoagulant tubes for flow cytometry (FCM) analysis and procoagulant separation tubes for the separation of serum. After centrifugation, the serum was subjected to ELISA and biochemical tests. The weights of the tumors, spleens, and thymuses were recorded after isolation. The tumor inhibition rate was calculated as (1 – a/b) × 100% (a indicates the weights of the tumors in the experimental groups; b represents the weights in the NS group). The spleen and thymus indices of the mice were calculated with the following formula: Organ index = spleen (thymus) weight/body weight ×100%. Polyoxymethylene (4%) was used to preserve the tumors for immunohistochemical tests.

2.7. ELISA and Biochemical Tests

The serum levels of TNF-α and IL-10 in tumor-bearing mice were detected with ELISA kits (Elabscience, Wuhan, China) according to the manufacturer’s instructions. Biochemical analysis was performed with the use of a fully automatic biochemical analyzer (HITACHI, Japan). Qualified operations were performed to test the levels of liver (alanine aminotransferase [ALT] and aspartic transaminase [AST]) and renal (creatinine [CREA] and blood urea [UREA]) function indices.

2.8. FCM Analysis

FCM analysis was performed for the tests of CD3, CD4, and CD8 positive cells, and the whole blood was mixed with the corresponding antibodies at the proper proportions. The CD3 antibody was labeled with FITC, the CD4 antibody was labeled with PE, and the CD8 antibody was labeled with APC. The mixture was kept in the dark for 20 min at room temperature. Lysing buffer (BD Biosciences, USA) was used to lyse red blood cells, and the reaction was completed at 37 °C for 20 min. After three washes with PBS (×200 g, 5 min), the cells were suspended in PBS for FCM analysis. A BD AccuriFM C6 Plus Flow Cytometer was used for FCM analysis.

2.9. Immunohistochemical Analysis

Paraffinized tumor sections were generated from the tumor tissues. Proliferation cell nuclear antigen (PCNA) staining was performed with the following procedure: dewaxing, antigen retrieval, blocking endogenous peroxidase, nonspecific binding blocking, primary PCNA antibody (Servicebio, China), incubation (4 °C, overnight), and secondary antibody incubation (50 min, room temperature). 3DHISTECH was used to visualize the sections stained with diaminobenzidine (DAB), and three random fields (400× magnification) from each image were obtained. The proliferation index was calculated as the ratio of PCNA-positive cells to total cells.

2.10. Statistical Analysis

The average value and variation in the quantitative data are expressed as the mean ± standard deviation (SD). One-way analysis of variance (ANOVA) was performed when the data were compared among more than two groups, and differences between two groups were further analyzed with Student–Newman–Keuls multiple comparison tests. Student’s t test was used for comparisons between two groups. p < 0.05 was considered to indicate statistical significance. All the statistical analyses were performed with IBM SPSS Statistics 21.

3. Results

3.1. Characteristics of PSP-AuNPs

The optical conditions for the synthesis of PSP-AuNPs were as follows: 1 mg/mL PSP in a 3.5 mL reaction mixture with 0.5 μL of HAuCl4 solution (1 M) were reacted at 80 °C for 15 min. From the UV–VIS results (Figure 1A), the characteristic absorption peak of PSP-AuNPs is between 540 and 550 nm, which is near the absorption peak of AuNPs. The TEM (Figure 1B) and AFM images (Figure 1C) demonstrated the successful synthesis of PSP-AuNPs, and the overall shape and surface morphology of PSP-AuNPs were different from those of AuNPs. The size (Figure 1D) and zeta (Figure 1E) distributions of the PSP-AuNPs determined by DLS were similar to normal distribution. The sizes of the PSP-AuNPs were mainly between 91 and 459 nm, and the zeta potentials were ranged from −35 to 10 mV. According to the EDX results, the percentages of C, O, and Au are 74.70%, 7.78%, and 17.52%, respectively. According to the FTIR analysis, the absorption peaks at 1699.53 cm–1, 3443.75, and 1539.33 cm–1 were ascribed to the stretching vibrations of C=O, −OH, and C=C, respectively. The stretching vibration of C–H accounted for the absorption peaks at 3188.18 and 2892.31 cm–1.

Figure 1.

Figure 1

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Characteristics of PSP-AuNPs. (A) UV–VIS images of PSP, AuNPs, and PSP-AuNPs. (B) TEM images of AuNPs and PSP-AuNPs. (C) AFM images of AuNPs and PSP-AuNPs. (D) Size and (E) zeta distributions determined by DLS. (F) EDX analysis. (G) FTIR results.

3.2. Concentrations of NO, TNF-α, and IL-12p70 in the Culture Media of RAW264.7 Cells

As demonstrated (Figure 2), the concentrations of NO (Figure 2A), TNF-α (Figure 2B), and IL-12p70 (Figure 2C) in the culture media of RAW264.7 cells stimulated with PSP and PSP-AuNPs were greater than those in the NC group and lower than those in the LPS group. The concentrations of NO, TNF-α, and IL-12p70 in the culture media of the PSP-AuNPs were greater than those in the PSP group at stimulation concentrations from 200 to 500 μg/mL. The concentrations of NO, TNF-α, and IL-12p70 reached their highest values at 400 μg/mL for PSP and 300 or 400 μg/mL for PSP-AuNPs.

Figure 2.

Figure 2

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Concentrations of NO (A), TNF-α (B), and IL-12p70 (C) in the culture medium of RAW264.7 cells. *p < 0.05, **p < 0.01, compared with the NC group; #P < 0.05, ##P < 0.01, compared with the PSP group.

3.3. The Viability of RAW264.7 and L02 Cells Treated with PSP-AuNPs

The viability of RAW264.7 cells (Figure 3A) was greater in the PSP and PSP-AuNP groups than in the NC group and lower than that in the LPS group, and the viability in the PSP-AuNP group was greater than that in the PSP group at concentrations from 200 to 500 μg/mL. The viability of L02 cells (Figure 3B) treated with PSP and PSP-AuNPs was greater than that of the ADM and NC groups, and the viability of the PSP-AuNP group was greater than that of the PSP group at all treated concentrations.

Figure 3.

Figure 3

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Viability of RAW264.7 (A) and L02 cells (B). *p < 0.05, **p < 0.01, compared with the NC group; #P < 0.05, ##P < 0.01, compared with the PSP group.

3.4. Biocompatibility and Toxicity of PSP-AuNPs In Vivo

The body weights of the mice in the PSP, PSP-AuNPs, and NS groups were increased with time and were decreased in the ADM and PSP-AuNPs+ADM groups (Figure 4A). The body weights of the PSP group were greater than those of the NS group and lower than those of the PSP-AuNP group. The body weights of the PSP-AuNPs + ADM group were greater than those of the ADM group and lower than those of the NS group. The serum levels of ALT, AST (Figure 4B), CREA, and UREA (Figure 4C) were lower in the PSP and PSP-AuNP groups than those in the NS group. Liver and renal function indices were lower in the PSP-AuNP group than those in the PSP group. The liver and renal function indices were lower in the PSP-AuNPs + ADM group than those in the ADM group. In addition, PSP-AuNPs increased the activities of the mice in the PSP-AuNPs and PSP-AuNPs + ADM groups.

Figure 4.

Figure 4

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Biocompatibility and toxicity of PSP-AuNPs in hepatic carcinoma tumor-bearing mice. (A) The trends in body weight. (B) The serum levels of liver function indices (ALT and AST). (C) The serum levels of renal function indices (CREA and UREA). *p < 0.05, **p < 0.01, compared with the NS group; #P < 0.05, ##P < 0.01, compared with the PSP or ADM group.

3.5. Immunoregulation of PSP-AuNPs In Vivo

The spleen (Figure 5A) and thymus (Figure 5B) indices, serum TNF-α levels (Figure 5C), and CD4+/CD8+ lymphocyte ratio (Figure 5E) were greater, and the serum IL-10 level (Figure 5D) was lower in the PSP-AuNP group than in the NS and PSP groups. Similarly, compared with those in the ADM group, the spleen and thymus indices, serum TNF-α levels, and CD4+/CD8+ lymphocyte ratio were greater, and the serum IL-10 concentration was lower in the PSP-AuNPs + ADM group.

Figure 5.

Figure 5

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Immunoregulation of PSP-AuNPs in vivo. (A) Spleen index. (B) Thymus index. (C) Serum TNF-α concentration. (D) IL-10 concentration. (E) CD4+/CD8+ lymphocyte ratio. (F) FCM analysis of CD4+ and CD8+ lymphocytes.

3.6. Antitumor Effects of PSP-AuNPs

The PSP-AuNP group demonstrated a greater tumor inhibition rate (Figure 6B), slower tumor growth (Figure 6C), and lower proliferation index (Figure 6D) than the PSP and NS groups did. The tumor inhibition rate was greater, tumor growth was slower, and the proliferation index was lower in the PSP-AuNPs + ADM group than those in the ADM group.

Figure 6.

Figure 6

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Antitumor effect of PSP-AuNPs. (A) Images of the isolated tumors. (B) Tumor inhibition rate. (C) Growth trends of tumor volumes. (D) Proliferation indices of tumor cells. (E) Images of PCNA staining (×400).

4. Discussion

In this study, we adopted a green method to synthesize gold nanoparticles from PSP to reduce the toxicity and increase the function of PSP. The results of the in vitro and in vivo experiments revealed that, compared with PSP, PSP-AuNPs could increase immunoregulation, anticancer effects, and biocompatibility.

The successful synthesis of PSP-AuNPs was primarily identified by the color change of the reaction solution from yellow to ruby red, then confirmed by the UV–VIS, TEM, AFM, DLS, EDX, and FTIR results. The sizes of PSP-AuNPs determined by TEM were smaller than those of AuNPs, whereas the characteristic absorption peak of AuNPs was lower than that of PSP-AuNPs, which might be attributed to the different syntheses process between PSP-AuNPs and AuNPs. The AFM results showed that the surface morphology of PSP-AuNPs were more irregular than that of AuNPs, which might be due to the combination of PSP in PSP-AuNPs. The sizes of PSP-AuNPs tested by DLS were larger than those tested by TEM, which might be due to the different degree of dispersion of PSP-AuNPs in the solution and the difference between these two kinds of test methods. Nanoparticles with sizes larger than 6–8 nm and negatively charged can be prevented from filtration by the kidneys.27,34 Form the DLS results, the sizes of the PSP-AuNPs were large enough, and the zeta potentials were mostly negative, which prevented filtration by the kidneys and allowed the accumulation of PSP-AuNPs in the liver. The EDX results demonstrated the presence of C, O, and Au in the PSP-AuNPs, and the percentages of C and O in PSP-AuNPs were very high, demonstrating the large amounts of PSP combined to AuNPs. The functional groups of C=O, −OH, C=C, and C–H identified by FTIR suggested the endogenous reducibility of PSP35 which laid the foundations of PSP-AuNPs function.

PSP-AuNPs demonstrated greater biocompatibility than PSP did: in vitro, the viabilities of RAW264.7 macrophages and L02 cells treated with PSP-AuNPs were greater than those treated with PSP; and in vivo, tumor-bearing mice treated with PSP-AuNPs had greater body weights, greater activity, and lower liver function (ALT and AST) and renal function (CREA and UREA) indices. Therefore, PSP-AuNPs could be safer than PSP for drug use. Immunoregulatory function of PSP-AuNPs in vitro was evaluated by testing the levels of immunoregulation factors including NO, TNF-α, and IL-12p70 secreted by RAW264.7 cells. The serum levels of TNF-α and IL-10, CD4+/CD8+ T lymphocyte ratio showed by FCM analysis, and spleen and thymus indices were tested to demonstrate the in vivo immunoregulatory function of PSP-AuNPs. NO might be considered as a potential therapeutic agent for cancer because it can regulate immunity through the activation of T lymphocytes.36,37 TNF-α functions in innate immunity and the lymphocyte-mediated adaptive immune response, and high-dose local administration of TNF-α results in powerful tumor necrosis.38,39 IL-12p70 originates from monocytes, and systemic administration of IL-12p70 has been demonstrated to have antitumor effects on various preclinical animal models as well as modest activity in cancer patients.40 IL-10 is secreted mainly by macrophages, and this cytokine can induce immunosuppression and tumor progression.41 Increased expression of IL-10 is correlated with poorer prognosis in several viral-correlated malignancies.42 The CD4+/CD8+ T lymphocyte ratio is often used to evaluate immune system function, and an increased ratio can predict a favorable treatment response.43 PSP-AuNPs exhibited greater immunoregulatory effects than PSP in vitro because of the greater expression levels of NO, TNF-α, and IL-12p70 in the culture medium of RAW264.7 macrophages treated with PSP-AuNPs. In vivo, the serum TNF-α concentration, CD4+/CD8+ T lymphocyte ratio, and spleen and thymus indices were greater, and the serum IL-10 concentration was lower in the PSP-AuNPs group than those in the PSP group, suggesting that the immunoregulation of PSP-AuNPs was greater. The biocompatibility and immunoregulation functions of PSP-AuNPs and PSP were tested on different concentrations to determine the appropriate concentrations of PSP-AuNPs for use. According to the study by Long T et al., PSP exerts tumor inhibitory effects through immunoregulation.44 The hepatic carcinoma tumor volumes grew more slowly, the tumor inhibition rate was greater, and the tumor cell proliferation rate demonstrated by PCNA staining of the tumor sections was lower in the PSP-AuNPs-treated group than those in the PSP group. Therefore, PSP-AuNPs demonstrated greater hepatic carcinoma tumor inhibition effect through immunoregulation than PSP did. In contrast to previous studies,30,45 we synthesized PSP-AuNPs from PSP with high a purity (>90%), and this strength facilitated studying the particular immunoregulation and anticancer function of PSP-AuNPs. We not only synthesized the nanoparticles successfully but also confirmed the anticancer effect on hepatic carcinoma of PSP-AuNPs through immunoregulation.

Interestingly, the tumor inhibition effect was greater in the PSP-AuNPs + ADM group than that in the ADM and PSP-AuNPs groups. In contrast to the destruction of immunoregulation, liver and renal functions caused by ADM, the combined use of PSP-AuNPs and ADM could help reverse some of those functions and protect the organs. As a result, the combined use of PSP-AuNPs and ADM could strengthen the anticancer effects of ADM and reduce the adverse effects.

5. Conclusions

The synthesis of PSP-AuNPs via a green method could increase the tumor inhibition effect of PSP in the treatment of hepatic carcinoma through enhancing immunoregulation. The combined use of PSP-AuNPs could reduce the adverse effects caused by chemotherapy and strengthen the anticancer effects. Our study has provided potential strategies for the future treatment of hepatic carcinoma.

Acknowledgments

The authors thank the Pediatric Medical Research Institute of Children’s Hospital Affiliated to Zhengzhou University and Academy of Medical Sciences of Zhengzhou University for providing laboratories and instruments to perform the experiments.

Data Availability Statement

The data underlying the article are available throughout the manuscript and supporting files.

This work was supported by Tianjian Laboratory of Advanced Biomedical Sciences.

The authors declare no competing financial interest.

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