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. 2022 Nov 23:13:1043945.
doi: 10.3389/fphar.2022.1043945. eCollection 2022.

Protective effects of mefunidone on ischemia-reperfusion injury/Folic acid-induced acute kidney injury

Jiajia Li  1   2   3 , Yupeng Jiang  1   2   3   4 , Qin Dai  1 , Yue Yu  1 , Xin Lv  1 , Yan Zhang  1 , Xiaohua Liao  1 , Liyun Ao  1 , Gaoyun Hu  5 , Jie Meng  2   6 , Zhangzhe Peng  1   2   3 , Lijian Tao  1   2   3 , Yanyun Xie  1   2   3
Affiliations

Protective effects of mefunidone on ischemia-reperfusion injury/Folic acid-induced acute kidney injury

Jiajia Li et al. Front Pharmacol. .

Erratum in

Abstract

Renal ischemia-reperfusion injury (IRI) is one of the most common causes of acute kidney injury (AKI). It poses a significant threat to public health, and effective therapeutic drugs are lacking. Mefunidone (MFD) is a new pyridinone drug that exerts a significant protective effect on diabetic nephropathy and the unilateral ureteral obstruction (UUO) model in our previous study. However, the effects of mefunidone on ischemia-reperfusion injury-induced acute kidney injury remain unknown. In this study, we investigated the protective effect of mefunidone against ischemia-reperfusion injury-induced acute kidney injury and explored the underlying mechanism. These results revealed that mefunidone exerted a protective effect against ischemia-reperfusion injury-induced acute kidney injury. In an ischemia-reperfusion injury-induced acute kidney injury model, treatment with mefunidone significantly protected the kidney by relieving kidney tubular injury, suppressing oxidative stress, and inhibiting kidney tubular epithelial cell apoptosis. Furthermore, we found that mefunidone reduced mitochondrial damage, regulated mitochondrial-related Bax/bcl2/cleaved-caspase3 apoptotic protein expression, and protected mitochondrial electron transport chain complexes III and V levels both in vivo and in vitro, along with a protective effect on mitochondrial membrane potential in vitro. Given that folic acid (FA)-induced acute kidney injury is a classic model, we used this model to further validate the efficacy of mefunidone in acute kidney injury and obtained the same conclusion. Based on the above results, we conclude that mefunidone has potential protective and therapeutic effects in both ischemia-reperfusion injury- and folic acid-induced acute kidney injury.

Keywords: acute kidney injury; chronic kidney disease; folic acid; mefunidone; renal ischemia-reperfusion injury.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Mefunidone ameliorated IRI-induced AKI (A) HE staining showed protective effect of mefunidone at various doses of 50 mg/kg, 75 mg/kg, 100 mg/kg, 125 mg/kg on renal tubular injury on day 2 after IRI modeling (×200). arrows for renal tubular damage. (B) The tubular injury scores of HE staining for kidney damage. (C) Serum creatinine (SCr) levels of mefunidone at various doses of 50 mg/kg, 75 mg/kg, 100 mg/kg, 125 mg/kg on renal tubular injury on day 2 after IRI modeling. (D) Blood urea nitrogen (BUN) levels of mefunidone at various doses of 50 mg/kg, 75 mg/kg, 100 mg/kg, 125 mg/kg on renal tubular injury on day 2 after IRI modeling. (E,F) Histological images of immunohistochemical staining with NGAL and evaluation of NGAL positive area in each group on day 2 after IRI modeling (×200). Mefunidone: 100 mg/kg. (G) Western blot analysis and quantitative data of NGAL in each group on day 2 after IRI modeling. Mefunidone: 100 mg/kg. Data represent mean ± SEM (n = 3–5). *p < 0.05, vs. Sham group; **p < 0.01, vs. Sham group; ***p < 0.001, vs. Sham group; #p < 0.05, vs. IRI group; ##p < 0.01, vs. IRI group; ###p < 0.001, vs. IRI group.
FIGURE 2
FIGURE 2
Mefunidone reduced IRI-induced oxidative stress accumulation. (A) DHE immunofluorescence staining for ROS detection in the renal tissues on day 2 after IRI modeling (×200). (B) Renal MDA level in the renal tissues on day 2 after IRI modeling. (C) RT-qPCR of CAT in the renal tissues on day 2 after IRI modeling. (D) RT-qPCR of mnSOD in the renal tissues on day 2 after IRI modeling. Mefunidone: 100 mg/kg. Data represent mean ± SEM (n = 3–5). *p < 0.05, vs. Sham group; **p < 0.01, vs. Sham group; ***p < 0.001, vs. Sham group; # p < 0.05, vs. IRI group; ## p < 0.01, vs. IRI group.
FIGURE 3
FIGURE 3
Mefunidone abated renal apoptosis (A) TUNEL staining in the renal tissues on day 2 after IRI modeling (×400). (B) Quantification of TUNEL positive cell count per scope. (C) Western blot analysis and quantitative data of cleaved-caspase3 in the renal tissues on day 2 after IRI modeling. (D,E) Flow cytometry and quantitative comparison were applied to detect apoptosis of HK-2 cells in H/R model with or without mefunidone. (F) Western blot analysis and quantitative data of cleaved-caspase3 in the H/R model of HK-2 cells with or without mefunidone. Dose of mefunidone in mice: 100 mg/kg; Dose of mefunidone in HK-2 cells: 80 μg/ml. Data represent mean ± SEM (n = 3–5). *p < 0.05, vs. Sham group; **p < 0.01, vs. Sham group; ***p < 0.001, vs. Sham group; # p < 0.05, vs. Modeling group; ## p < 0.01, vs. Modeling group; ### p < 0.001, vs. Modeling group.
FIGURE 4
FIGURE 4
Mefunidone protected IRI-induced mitochondrial damage and complex Ⅲ and Ⅴ of mitochondrial electron transport chain complex (A) Electron microscope analysis of mitochondria in the renal tissues on day 2 after IRI modeling (×8,000). (B,C) Western blot analysis and quantitative data of bax and bcl2 in the renal tissues on day 2 after IRI modeling. (D,E) Western blot analysis and quantitative data of mitochondrial electron transport chain complex in the renal tissues on day 2 after IRI modeling. Mefunidone: 100 mg/kg. Data represent mean ± SEM (n = 3). *p < 0.05, vs. Sham group; **p < 0.01, vs. Sham group; # p < 0.05, vs. IRI group.
FIGURE 5
FIGURE 5
Mefunidone reduced H/R-induced mitochondrial damage and complex Ⅲ and Ⅴ of mitochondrial electron transport chain complex in HK-2 cells (A,B) Western blot analysis and quantitative data of bax and bcl2 in the H/R model of HK-2 cells with or without mefunidone. (C) Flow cytometry and quantitative comparison were applied to test MMP of HK-2 cells in the H/R model of HK-2 cells with or without mefunidone. (D,E) Western blot analysis and quantitative data of mitochondrial electron transport chain complex in the H/R model of HK-2 cells with or without mefunidone. Mefunidone: 80 μg/ml. Data represent mean ± SEM (n = 3–5). *p < 0.05, vs. Control group; **p < 0.01, vs. Control group; ***p < 0.001, vs. Control group; ****p < 0.0001, vs. Control group; # p < 0.05, vs. H/R model group; ## p < 0.01, vs. H/R model group.
FIGURE 6
FIGURE 6
Mefunidone ameliorated FA-induced AKI (A) HE staining showed protective effect of mefunidone on renal tubular injury on day 2 after FA modeling. arrows for renal tubular damage. (×200). (B) The tubular injury scores of HE staining for kidney damage. (C) Serum creatinine (SCr) levels on renal tubular injury on day 2 after FA modeling. (D) Blood urea nitrogen (BUN) levels on renal tubular injury on day 2 after FA modeling. (E) Western blot analysis and quantitative data of NGAL in each group on day 2 after FA modeling. (F) Western blot analysis and quantitative data of cleaved-caspase3 in each group on day 2 after FA modeling. Mefunidone: 100 mg/kg. Data represent mean ± SEM (n = 3–6). *p < 0.05, vs. Sham group; **p < 0.01, vs. Sham group; ***p < 0.001, vs. Sham group; # p < 0.05, vs. FA group; ## p < 0.01, vs. FA group.
FIGURE 7
FIGURE 7
Potential mechanisms of mefunidone for the treatment of IRI/FA-induced AKI. Mefunidone may improve IRI-induced AKI by inhibiting oxidative stress, reducing apoptosis and protecting mitochondria.
See this image and copyright information in PMC

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