Berberine protects renal tubular cells against hypoxia/reoxygenation injury via the Sirt1/p53 pathway
Berberine protects renal tubular cells against hypoxia/reoxygenation injury via the Sirt1/p53 pathway
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小檗碱通过 Sirt1/p53 通路保护肾小管细胞免受缺氧/复氧损伤
DOI：
10.1007/s11418-018-1210-1
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发表时间：
2018-06-01
影响因子：
3.3
通讯作者：
Yu, Wenli
中科院分区：
文献类型：
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作者：
Lin, Yuanbang;Sheng, Mingwei;Yu, Wenli
小檗碱（BBR）已被证明对肾缺血/再灌注损伤有保护作用，然而，其潜在的分子机制在很大程度上是未知的。在本研究中，我们研究了沉默信息调节因子1（Sirt 1）/p53在BBR对大鼠肾小管上皮细胞（NRK-52 E细胞）缺氧/复氧（H/R）介导的线粒体功能障碍的保护作用中的作用。NRK-52 E细胞在H/R之前用靶向Sirt 1的小干扰RNA（Sirt 1-siRNA）和BBR预处理。通过CCK 8测定和氧化参数检测来评估细胞损伤。流式细胞术和Hoechst 33258染色检测细胞凋亡率。Western blotting检测凋亡标志物Sirt 1、p53的表达及p53的易位。使用免疫沉淀法检测Sirt 1引起的核p53脱乙酰化。与H/R组相比，BBR预处理增加细胞活力，抑制线粒体氧化应激和凋亡。Sirt 1蛋白表达也随着p53的减少而沿着增强。此外，BBR预处理的NRK-52 E细胞中p53的核转位及其乙酰化均受到抑制。然而，Sirt 1的敲除抵消了BBR的肾保护作用。BBR预处理通过调节Sirt 1/p53通路保护大鼠肾小管上皮细胞对抗缺氧/复氧诱导的线粒体功能障碍
Berberine (BBR) has been demonstrated to protect against renal ischemia/reperfusion injury; however, the underlying molecular mechanism is largely unknown. In the present study, we examined the role of silent information regulator 1 (Sirt1)/p53 in the protective effect of BBR on hypoxia/reoxygenation (H/R)-mediated mitochondrial dysfunction in rat renal tubular epithelial cells (NRK-52E cells). NRK-52E cells were preconditioned with small interfering RNA targeting Sirt1 (Sirt1-siRNA) and BBR before subjected to H/R. Cell damage was assessed by CCK8 assay and detection of oxidative parameters. The apoptotic rate was determined by flow cytometry and Hoechst 33258 staining. The expression of apoptotic markers, Sirt1, p53 and the translocation of p53 were examined by Western blotting assay. Nuclear p53 deacetylation by Sirt1 was detected using immunoprecipitation. Compared with the H/R group, BBR pretreatment increased cell viability and inhibited mitochondrial oxidative stress and apoptosis. Protein expression of Sirt1 was also enhanced along with a reduction of p53. Furthermore, both nuclear translocation of p53 and its acetylation were inhibited in NRK-52E cells pretreated with BBR. However, the knockdown of Sirt1 counteracted the renoprotection of BBR. BBR preconditioning protects rat renal tubular epithelial cells against H/R-induced mitochondrial dysfunction via regulating the Sirt1/p53 pathway.