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. 2026 Sep 1;109(3):00368504261486989. doi: 10.1177/00368504261486989

Involvement of NOD1/NF-κB/MAPK pathway in the protective effect of procyanidin B2 against methotrexate-induced nephrotoxicity

Ahmed A Altuwaijri 1, Ahlam M Alhusaini 1,✉, Wedad S Sarawi 1, Reema A Alabdani 1, Reyouf I Alnajashi 1, Iman H Hasan 1, Hanaa M Altuwaijri 2, Qamraa H Alqahtani 1, Dalal A Alkhelb 1, Sahar S Alotaibi 1, Nadrah S Alamri 1, Raeesa A Mohammad 3, Ebtesam N Alsultan 4, Shaikha M Alsaif 5, Ebtesam A Aljasas 5
PMCID: PMC13535005  PMID: 42677976

Abstract

Objective

Methotrexate (MTX) is a very effective agent used to treat rheumatoid arthritis and malignant diseases. Despite its usefulness, the long-term use of MTX is associated with multiorgan toxicities, including kidney injury. Thus, there is a necessity to explore a renoprotective agent with low side effects. Procyanidin B2 (PCB2), a natural flavonoid, possesses antioxidant and anti-inflammatory activities. This study aims to investigate the potential protective efficacy of PCB2 against MTX-induced nephrotoxicity.

Methods

Rats were allocated into four groups: control group, PCB2 group (40 mg/kg, p.o), MTX-intoxicated group (20 mg/kg, i.p.), and PCB2+MTX group. PCB2 was administered daily for ten days, while MTX was injected as a single dose on day 8. Oxidative stress and inflammatory biomarkers in kidney tissues were assessed using biochemical, histological, and immunohistochemical examinations.

Results

MTX markedly elevated the levels of kidney malondialdehyde (MDA), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6), while glutathione (GSH) level and superoxide dismutase (SOD) activity were significantly reduced. In addition, rats intoxicated with MTX exhibited positive and strong immunoreactions for nucleotide-binding oligomerization domain 1 (NOD1), nuclear factor-kappa B (NF-κB), and mitogen-activated protein kinase (MAPK). The administration of PCB2 significantly prevented tissue injury and restored the altered parameters.

Conclusions

PCB2 reduced oxidative stress and inflammatory responses, indicating potential protective effects against MTX-induced kidney damage.

Keywords: methotrexate, nephrotoxicity, procyanidin B2, NOD1, NF-κB, p38MAPK

1. Introduction

Methotrexate (MTX) is a potent dihydrofolate reductase (DHFR) inhibitor, triggering a reduction of intracellular tetrahydrofolate, which is the active form of folic acid. 1 This action makes MTX a very effective drug to treat rheumatoid arthritis and malignant diseases. 1 Although it is a highly effective drug, preclinical and clinical studies reported that high doses of MTX are linked with kidney injury.2,3 Even with close monitoring of blood MTX levels and using different preventive measures, 1%-48% of patients receiving MTX develop dose-dependent acute kidney injury.4–6 Accordingly, MTX-induced nephrotoxicity is a major limitation to its long-term use in patients.

The exact mechanism involved in MTX-induced nephrotoxicity is still not fully elucidated, but it could be attributed to the accumulation of MTX metabolite, 7-hydroxy MTX, in renal tubules. 7 Besides, the generation of reactive oxygen species (ROS), the release of inflammatory mediators, and the initiation of apoptosis are suggested mechanisms underlying MTX kidney injury.8,9 ROS production can initiate oxidative stress, which triggers the activation of proinflammatory pathways, including nuclear factor kappa-B p65 (NF-κB) and mitogen-activated protein kinase P38 (MAPK), both of which are upstream regulators in the release of inflammatory cytokines such as tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6). 10 Moreover, the activation of nucleotide-binding oligomerization domain 1 (NOD1) can induce inflammatory as well as stress cascades, mainly activating MAPK and NF-κB pathways. 11 Therefore, maintaining the balance of redox and inflammatory processes is necessary to address MTX-induced renal toxicity.

In this context, Procyanidin B2 (PCB2), which is found in the extracts of proanthocyanidin, has been found to exhibit antioxidant, anti-inflammatory, and antiapoptotic activities. It has been reported that PCB2 could inhibit NF-κB and prevent cell injury in human endothelial cells induced by oxidized low-density lipoprotein. 12 Additionally, PCB2 exhibited a protective activity against MTX-induced liver damage through reducing oxidative stress and downregulating inflammatory mediators, including TLR4, NF-κB, TNF-α, and IL-6. 8

Thus, the current study examined for the first time the protective effect of PCB2 against MTX-induced kidney injury in rats, focusing on its antioxidant and anti-inflammatory effects. Mechanistically, we investigated the role of the NOD1/NF-κB/MAPK pathway, providing molecular insight into the modulatory mechanisms of PCB2.

2. Materials and methods

2.1. Chemicals

Methotrexate (MTX) isotonic liquid vials (NDC 61703-350-38; Hospira, Illinois, USA) were obtained from King Saud Hospital, Unaizah, Saudi Arabia. Procyanidin B2 (PCB2) (Cat. No. BP1147) was purchased from Biopurify Phytochemicals Ltd (Chengdu, China). antibody against NOD1 (Cat. No. K002564P) was purchased from Solarbio Life Sciences Company. Beijing, China. Antibodies against NF-κB (Cat. No. 8242S) and MAPK (Cat. No. 9212S) were purchased from Cell signalling, Boston, USA. Biotinylated Anti-Rabbit Secondary Antibody (Cat. No. 31820) was purchased from Thermo Fisher Scientific.

2.2. Experimental design

A total of 32 healthy male adult Wistar Albino rats, with weights ranging from 200 to 250 g, were obtained from the Animal Care Centre at the Pharmacy College of King Saud University, Riyadh, Saudi Arabia. The sample size was calculated according to the method of “resource equation” 13 : E = Total number of animals − Total number of groups. So, E = 32 – 4 = 28 which is more than 20; hence sample size in this experiment is within the acceptable limit.

The rats were housed in standard polypropylene ventilated cages (four rats/cage) with free access to water and chows. The animals were maintained in a controlled environment at 25°C under a 12-h dark/light cycle for one week to acclimatize before the experiments. The animals have received adequate care according to the “Guide for the Care and Use of Laboratory Animals”. 14 The experiment was authorized by the Research Ethics Committee at King Saud University (Ethics Reference No: KSU‐ SE-23-15) on (18/4/2023), and reported in accordance of the ARRIVE guidelines. 15

The rats were randomly divided into four groups, 8 rats/each. Group I: Control group; rats administered an equivalent volume of carboxymethylcellulose (CMC) 1% orally. Group II: PCB2-control group; rats received a daily oral dose of PCB2 (40 mg/kg dissolved in CMC 1%) for ten days. 16 Group III: MTX group; rats received CMC 1% orally and MTX in a single dose (20 mg/kg, i.p.) 17 on day 8. Group IV: MTX+PCB2 group; rats received PCB2 for ten days and a single MTX dose on day 8. The doses of PCB2 and MTX were selected according to previously published studies.16,17

On day 11, the rats were euthanized using pure compressed CO2 gas approximately 99.99%, followed by decapitation. Blood samples were collected for serum separation by centrifugation at 3000 rpm at 4°C for 15 minutes. Kidney samples were collected and kept in -80 °C for biochemical analysis, and the other kidney samples were kept in 10% formalin for histological and immunological examinations. All samples were anonymously coded to prevent bias.

2.3. Biochemical assays

Serum creatinine level was assessed using a colorimetric assay kit (Cat. No. 033K-240) which purchased from (United Diagnostic Industry kits, Dammam, Saudi Arabia). The serum levels of uric acid (Cat. No. EEA053) and urea (Cat. No. EIABUN) were measured using colorimetric assay kits (Invitrogen, Thermo Fisher Scientific, Massachusetts, USA).

Renal malondialdehyde (MDA) levels were measured using Solarbio Life Sciences kits (Cat. No. BC0025). In addition, reduced glutathione (GSH) levels and superoxide dismutase (SOD) activity in kidney tissues were assessed using Solarbio Life Sciences kits (Cat. No. CB1175) and (Cat. No. BC0175), respectively.

Renal inflammatory biomarkers TNF-α and IL-6 levels, were quantified using Solarbio Life Sciences ELISA kits (Cat. No. SEKR-0009 and SEKR-0005, respectively), following the manufacturer’s instructions.

2.4. Histopathological and immunohistochemistry examination

Fixed tissue specimens were processed and embedded in paraffin to obtain sections of 5 µm thickness. For histopathological evaluation, sections were stained with hematoxylin and eosin (H&E) to assess structural alterations in the renal glomeruli and tubules. The quantitative histopathological analysis was performed using the images at x200 magnification, and the percentage of injured glomeruli in all groups was normalized relative to the control group. The qualitative and quantitative analyses of histopathological images were blindly performed.

For immunohistochemical (IHC) analysis of NOD1, NF-κB, and MAPK, sections were deparaffinized in xylene and rehydrated through a descending ethanol series. Antigen retrieval was achieved by heating the slides in 10 mM citrate buffer (pH 6.0) for 6 minutes. To minimize background interference, endogenous peroxidase activity was quenched with 3% hydrogen peroxide, followed by a 30-minute incubation in a 5% blocking buffer to prevent non-specific protein binding.

The sections were then incubated overnight at 4°C with primary antibodies against NOD1, NF-κB, and p38MAPK as (1:50) in blocking buffer. After washing with phosphate-buffered saline (PBS), the slides were treated with the secondary antibody. The localized expression of target proteins was visualized using 3,3′-Diaminobenzidine (DAB) as the chromogen. Finally, the sections were counterstained with hematoxylin, dehydrated, and mounted for microscopic examination.

2.5. Statistical analysis

The Shapiro–Wilk normality test was used to assess whether the data were normally distributed and to determine the appropriate statistical tests. Statistical analysis was performed using one-way analysis of variance (ANOVA), followed by Tukey’s post hoc test for multiple comparisons. All analyses were conducted using GraphPad Prism 9 software. Data are presented as mean ± SEM, and p-value of < 0.05 was considered statistically significant.

3. Results

3.1. PCB2 restored the serum levels of creatinine, uric acid, and urea

Figure 1(a), (b), and (C) show a significant elevation in serum levels of creatinine, uric acid, and urea in rats intoxicated with MTX compared to the control group (p < 0.0001, η2 = 0.6797, 0.7768 and 0.8063, respectively). However, treatment with PCB2 significantly restored their levels compared to the MTX-intoxicated group (Figure 1(a),(b), (c)).

Figure 1.

Figure 1.

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Effects of PCB2 on serum levels of (A) creatinine, (B) uric acid, and (C) urea in MTX-induced nephrotoxicity in rats. Data are expressed as mean ± SEM. *** p < 0.001, **** p < 0.0001.

3.2. PCB2 reduced oxidative stress in renal tissues

As seen in Figure 2(a), MTX caused a marked elevation in the renal MDA level compared with the control group (p < 0.0001, η2 = 0.8656), while its level markedly reduced with the administration of PCB2 (p < 0.0001). In addition, rats treated with MTX exhibited a significant reduction in the levels of endogenous antioxidants, GSH and SOD (p < 0.0001, η2 = 0.7936 and 0.8910, respectively, Figure 2(b) and (c)). On the other hand, the intervention of PCB2 significantly restored GSH level (p < 0.01) and SOD (p < 0.0001) activity in renal tissues compared to the MTX-intoxicated group (Figure 2(b) and (c)).

Figure 2.

Figure 2.

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Effects of PCB2 on the levels of (A) MDA, (B) GSH, and (C) SOD in MTX-induced nephrotoxicity in rats. Data are expressed as mean ± SEM. ** p < 0.01, **** p < 0.0001.

3.3. PCB2 downregulated the expression levels of TNF-α and IL-6 in renal tissues

The expression levels of pro-inflammatory cytokines TNF-α and IL-6 were significantly elevated in the MTX-intoxicated group compared to the control group (p < 0.0001 η2 = 0.9532 and 0.9465, respectively, Figure 3(a) and (b)). However, administration of PCB2 caused a substantial reduction in TNF-α and IL-6 levels compared to the MTX-intoxicated group (p < 0.0001) (Figure 3(a) and (b)).

Figure 3.

Figure 3.

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Effects of PCB2 on the expression levels of (A) TNF-α and (B) IL-6 in MTX-induced nephrotoxicity in rats. Data are expressed as mean ± SEM. **** p < 0.0001.

3.4. PCB2 improved the morphological alterations in kidney tissues induced by MTX

The kidney sections obtained from the control and PCB2-treated rats (Figure 4 (a) and (b)) exhibited normal kidney tubules and corpuscles. The kidney section from MTX-administered rats displayed degenerated renal corpuscles with completely damaged glomeruli (Figure 4(c)). However, PCB 2 improved the histopathological changes induced by MTX overdose and restored the normal architecture (Figure 4(d)). Moreover, as illustrated in Figure 4(e), kidney sections from MTX-intoxicated group showed a significant increase in the injured glomeruli in comparison to normal control group (p < 0.0001). While the group treated with (MTX+PCB2) exhibited a marked reduction in the injured glomeruli when compared to MTX group (p < 0.0001).

Figure 4.

Figure 4.

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Photomicrograph of H&E-stained kidney sections, scale bar; 50μm. (A &B) are Kidney section from control and PCB2 administered rats, showing normal kidney tubules (arrows) and renal corpuscles (stars). (C) Kidney section from MTX administered rat shows apparently normal tubules (arrow), while renal corpuscles are markedly degenerated with completely destructed glomeruli (bent arrows). (D) Kidney section from rat received MTX and PCB2, show apparently normal tubules (arrows) and renal corpuscles (stars). (E) Percentage of injured glomeruli in all treated groups relative to the control. Data are expressed as mean ± SEM. **** p < 0.0001.

3.5. PCB2 reduced the positive immune reaction of NOD1, NF-kB, and MAPK in kidney tissues induced by MTX

Figure 5(a) and (b) showed mild immune reactions of NOD1 in both renal glomerulus and tubular epithelium in kidney tissues obtained from control and PCB2-treated rats. However, the kidney section from rat exposed to MTX showed a strong immunoreactivity of NOD1 in epithelium of the glomerulus and most of the renal tubule’s epithelium (Figure 5(c)). The immune reaction of NOD1 was reduced in renal tubules and glomeruli of kidney tissues obtained from rat received co-treatment of MTX and PCB2 (Figure 5(d)).

Figure 5.

Figure 5.

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Kidney sections immuno-stained with NOD1, NF-κB, and p38MAPK. Scale bar; 50μm. (A, E, and I) Control sections showing mild immune reaction in renal glomeruli (short arrow) and tubular epithelium (long arrow). (B, F, and J) Kidney sections from rat received PCB2, showing also mild immune positivity of renal glomerulus (short arrow) and epithelium in renal tubules (long arrow). (C, G, and K) Kidney sections from rats exposed to MTX showing strong immune reactions of epithelium of the glomerulus (short arrow) and most of the renal tubule’s epithelium (long arrows). (D, H, and L) Kidney sections from rats received MTX and PCB2 showing moderate immune positivity in renal glomeruli (short arrow) and tubules (long arrow).

Likewise, the administration of high dose of MTX caused a strong positive immune reaction of NF-κB of the epithelium of the renal corpuscle and most renal tubules, as well as interstitial cells (Figure 5(g)) compared to the control and PCB2-treated groups (Figure 5(e) and (f)). The kidney sections from rats treated with MTX and PCB2 exhibited moderate immunoreactivity of NF-κB in both renal glomerulus and tubules (Figure 5(h)).

As presented in Figure 5(k), kidney section from rat exposed to MTX showed strong immune positivity of p38MAPK in epithelium of the renal corpuscle and most renal tubules compared to sections extracted from control and PCB2-treated rats (Figure 5(i) and (j)). However, the kidney sections collected from rat received MTX and treated with PCB2 showed a moderate immune reaction of p38MAPK in both renal glomerulus and tubules (Figure 5(l)) compared to MTX-intoxicated rats (Figure 5(k)).

4. Discussion

In this study, we showed that PCB2 mitigates nephrotoxicity induced by MTX via modulation of oxidative stress, inflammation, and the NOD1/NF-κB/MAPK pathway. In the male adult Wistar Albino rats, a single intraperitoneal dose of MTX (20 mg/kg) increased the markers of nephrotoxicity, including uric acid, creatinine, and urea. Moreover, it increased oxidative stress and inflammation, as indicated by elevated MDA and depletion of GSH and SOD, along with elevated levels of TNF-α and IL-6. However, treatment with PCB2 attenuated the increased uric acid, creatinine, and urea in the MTX-treated rats. In addition, it restored GSH level and SOD activity, and reduced MDA, TNF-α, and IL-6 levels. Histological analysis showed that MTX injection resulted in degeneration of renal corpuscles and disruption of glomeruli, an effect that was efficiently reversed by PCB2 treatment. In addition, MTX was found to increase the protein expression of NF-κB, MAPK and NOD1 in the kidney. However, the administration of PCB2 reduced the activation of NF-κB, MAPK, and NOD1 associated with MTX treatment.

In the present study, MTX increased oxidative stress in the kidney, as demonstrated by elevated MDA and depletion of GSH, SOD, and CAT. Consistent with the results of this study, previous studies have reported that MTX increases ROS and lipid peroxidation, manifested by markers such as MDA, while depleting antioxidant defenses like GSH and SOD in different organs, including the kidney.18–21 MTX disrupts the balance between oxidant production and antioxidant defense systems via multiple biochemical pathways. 22 For example, MTX inhibits NADP-dependent dehydrogenases and NADP malic enzymes, reducing NADPH production and consequently impairing GSH regeneration.22,23 In addition, inhibition of dihydrofolate reductase promotes nitric oxide synthase uncoupling, leading to superoxide generation.24–26 MTX also increases xanthine oxidase activity, further enhancing ROS production. 23 Collectively, these mechanisms partially explain the elevated oxidative stress that contributes to nephrotoxicity in this model.

However, PCB2 reduced oxidative stress in the present study, evidenced by decreased MDA and increased GSH and SOD. PCB2 has been reported to have antioxidant activity through direct scavenging of ROS and enhancement of endogenous antioxidant systems. Consistently, pre-treatment with PCB2 reduced hepatic MDA and increased the activity of SOD, CAT, and GSH in a mouse model of acute liver injury. 27 Moreover, PCB2 restored the antioxidant capacity of GSH and SOD while decreasing hepatic MDA in MTX-injured rats. 8 In vitro, PCB2 decreased intracellular ROS and MDA levels while increasing SOD and glutathione peroxidase (GPx) activities in MCF-7 cells exposed to H2O2. 28 In addition, PCB2 promotes the nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2). Activated Nrf2 upregulates the synthesis of antioxidant enzymes, including oxygenase-1 and NADPH quinone oxidoreductase 1.29–31 These findings support the notion that PCB2 has an antioxidant property that contributes to kidney protection.

In addition to its antioxidant activity, PCB2 exhibited an anti-inflammatory effect through modulating the protein levels of the pro-inflammatory cytokines TNF-α and IL-6. In the current study, we showed that MTX upregulated the expression of TNF-α and IL-6 in kidney tissues compared to control group, suggesting that MTX activates innate immune and downstream inflammatory signaling pathways. Also, we showed that the rats treated with PCB2 exhibited reduced levels of TNF-α and IL-6 compared with MTX-intoxicated rats. Previous study has reported that MTX-induced nephrotoxicity is associated with increased renal expression of TNF-α and IL-6. 32 In accordance with our results, PCB2 suppressed the TLR4/NF-κB signaling pathway, resulting in lower production of inflammatory mediators such as TNF-α, IL-1β, and IL-6 in MTX-induced liver toxicity. 8 These findings suggest that the protective effects of PCB2 include the modulation of inflammatory signaling through TNF-α and IL-6.

To further evaluate the anti-inflammatory effectiveness of PCB2, immunohistochemical analysis was conducted. We showed that a high dose of MTX induced a positive immunoreactivity for NOD1, NF-κB, and MAPK compared to the control group. However, treatment with PCB2 reduced the immune reaction of NOD1, NF-κB, and MAPK, confirming its anti-inflammatory activity. Importantly, we proposed that PCB2 protects against kidney injury by modulating the NOD1 activity. NOD1 is a cytosolic pattern recognition receptor that senses cellular stress and initiates inflammatory responses.33–35 Activated NOD1 upregulates downstream pathways such as NF-κB and MAPK.34,35 The activation of NF-κB and MAPK signaling pathways induces the secretion of the pro-inflammatory cytokines, including TNF-α and IL-6.34,36 Moreover, NF-κB and MAPK promote apoptosis, fibrosis, and immune cell infiltration in kidney injury.34,37–41 In Consistent with our findings, previous studies have shown that MTX-induced nephrotoxicity involves activation of the NF-κB and p38MAPK signaling pathways, resulting in increased oxidative stress, inflammation, and apoptosis. 37 Together, these findings support a central role for the NF-κB and MAPK signaling pathways in the pathogenesis of MTX-induced renal injury. Importantly, our findings further suggest that modulation of NOD1 may contribute to the protective mechanism of PCB2 against MTX-induced nephrotoxicity.

Although this study showed that PCB2 has a protective effect against MTX-induced nephrotoxicity, several limitations should be acknowledged. First, the small sample size may limit the generalizability of the findings. Second, a single dose of MTX was used, which may not reflect their use in the clinical setting. Third, short duration (10 days) may not reflect the long-term effects of MTX and therefore the protective effect of PCB2. Moreover, we recognize the importance of incorporating additional molecular techniques, such as RT-PCR, Western blotting, or phosphorylation assays to establish more comprehensive mechanistic understanding aa well as to confirm the involvement of the NOD1/NF-κB/MAPK pathway. Addressing these limitations in the future will further strengthen the translational relevance of the present finding.

In conclusion, MTX caused nephrotoxicity by inducing oxidative stress and inflammatory responses in rats. Nephrotoxicity was attenuated by PCB2 via balancing redox reaction and modulating the NOD1/NF-κB/MAPK axis. These findings highlight the therapeutic potential of PCB2 as a supportive strategy against MTX-induced nephrotoxicity. Nevertheless, the outcomes of the current study warrant further investigation to determine the human dose, further molecular mechanisms, pharmacokinetic, safety, and its clinical outcomes.

Supplemental material

Supplemental material - Involvement of NOD1/NF-κB/MAPK pathway in the protective effect of procyanidin B2 against methotrexate-induced nephrotoxicity
sj-pdf-1-sci-10.1177_00368504261486989.pdf (14.7MB, pdf)

Supplemental material for Involvement of NOD1/NF-κB/MAPK pathway in the protective effect of procyanidin B2 against methotrexate-induced nephrotoxicity in Ahmed A. Altuwaijri, Ahlam M. Alhusaini, Wedad S. Sarawi, Reema A. Alabdani, Reyouf I. Alnajashi, Iman H. Hasan, Hanaa M. Altuwaijri, Qamraa H. Alqahtani, Dalal A. Alkhelb, Sahar S. Alotaibi, Nadrah S. Alamri, Raeesa A. Mohammad, Ebtesam N. Alsultan, Shaikha M. Alsaif, Ebtesam A. Aljasas in Science Progress.

Author contributions: Conceptualization: Alhusaini A. and Sarawi W.; Funding acquisition: Alhusaini A.; Resources: Alhusaini A., Sarawi W., Alotaibi S. Alsultan E., Alsaif S., Aljassas E.; Methodology: Alhusaini A., Sarawi W., Hasan I., Alabdani R., Alnajashi R., Alotaibi S. Altuwaijri H.; Formal analysis: Altuwaijri A., Hasan I., Sarawi W., Alamri N.; Writing - original draft: Alhusaini A., Altuwaijri A., Sarawi W., Alamri N., Hasan I., Alqahtani Q., Alkhelb D. All authors have read and agreed to the final version of the manuscript.

Funding: The authors disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This research was funded by the Ongoing Research Funding Program (ORF-2026-822), King Saud University, Riyadh, Saudi Arabia.

The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Supplemental material: Supplemental material for this article is available online.

ORCID iDs

Ahlam M. Alhusaini https://orcid.org/0000-0003-2905-8882

Iman H. Hasan https://orcid.org/0000-0001-7311-1412

Data Availability Statement

All supplementary data is available on request from the corresponding author.*

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Associated Data

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Supplementary Materials

Supplemental material - Involvement of NOD1/NF-κB/MAPK pathway in the protective effect of procyanidin B2 against methotrexate-induced nephrotoxicity
sj-pdf-1-sci-10.1177_00368504261486989.pdf (14.7MB, pdf)

Supplemental material for Involvement of NOD1/NF-κB/MAPK pathway in the protective effect of procyanidin B2 against methotrexate-induced nephrotoxicity in Ahmed A. Altuwaijri, Ahlam M. Alhusaini, Wedad S. Sarawi, Reema A. Alabdani, Reyouf I. Alnajashi, Iman H. Hasan, Hanaa M. Altuwaijri, Qamraa H. Alqahtani, Dalal A. Alkhelb, Sahar S. Alotaibi, Nadrah S. Alamri, Raeesa A. Mohammad, Ebtesam N. Alsultan, Shaikha M. Alsaif, Ebtesam A. Aljasas in Science Progress.

Data Availability Statement

All supplementary data is available on request from the corresponding author.*

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