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. 2018 Nov;40(1):498-505.
doi: 10.1080/0886022X.2018.1497517.

Effect of apigenin on apoptosis induced by renal ischemia/reperfusion injury in vivo and in vitro

Xiao Wang  1   2   3 , Wei Wang  1   2 , Jian-Zhong Wang  1 , Cheng Yang  1 , Chao-Zhao Liang  1   2
Affiliations

Effect of apigenin on apoptosis induced by renal ischemia/reperfusion injury in vivo and in vitro

Xiao Wang et al. Ren Fail. 2018 Nov.

Abstract

Objectives: This study aims to investigate the effects and molecular mechanisms of apigenin (ApI) on renal ischemia/reperfusion (I/R) injury in vivo and in vitro.

Methods: In vivo, the left renal artery was clamped for 45 min and the right kidney was removed to study renal I/R injury on Sprague-Dawley (SD) rats. ApI was injected at 60 min before renal ischemia. In vitro, renal tubular epithelial cells (HK-2) were pretreated with or without ApI (20 uM) for 60 min and then treated with hypoxia/reoxygenation (H/R). Renal function, histology, cells apoptosis, and cell viability were tested. Furthermore, the potential molecular mechanisms were assessed.

Results: ApI pretreatment could significantly alleviated the renal function and the pathological damage as well as cells apoptosis after I/R injury. Meanwhile, ApI treatment protects H/R induced HK-2 cell apoptosis in vitro. The results of Western blot showed that ApI significantly increased the expressions of B-cell lymphoma 2 (Bcl-2) and phosphor-AKt (p-AKt), Phosphoinositide 3-kinase (PI3K), while down-regulated the expressions of Caspase3 and Bax induced by H/R injury.

Conclusions: ApI pretreatment can protect renal function against I/R injury and prevent renal tubular cells from apoptosis in vivo and in vitro which might through PI3K/Akt mediated mitochondria-dependent apoptosis signaling pathway.

Keywords: AKt; Apigenin; Bcl-2; apoptosis; renal ischemia/reperfusion (I/R) injury.

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Figures

Figure 1.
Figure 1.
The Scr (A) and BUN (B) values measured after 45 min of ischemia followed by 24 h of reperfusion. #p < .05 vs. sham group, *p < .05 vs. IR group.
Figure 2.
Figure 2.
(A) Representative micrographs of H&E staining and (B) Quantitative damage score of H&E staining. #p < .05 vs. sham group, *p < .05 vs. IR group, Scale bar =100 µm.
Figure 3.
Figure 3.
(A) Representative micrographs of Tunnel staining. (B) #p < .05 vs. sham group, *p < .05 vs. IR group, Scale bar =100 µm.
Figure 4.
Figure 4.
(A) CCK-8 method was employed to detect cell viability after the treatment. (B) Representative images of FACS to measure apoptosis of HK-2 cells after the treatment. (C) Quantitation of apoptotic cells in 10000 cells. #p < .05 vs. Control group, *p < .05 vs. H/R group.
Figure 5.
Figure 5.
Western blot analysis of Caspase-3 and Cleaved-caspase3 (A), Bcl-2 and Bax (B), p-AKT and AKT (C). (B,D,E,F,G) and PI3K (H,I). Semi-quantitative analysis of western blot for different proteins. *p < .05 vs. Control group, #p < .05 vs. H/R group.
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