丙酮酸激酶通过靶向Bcl-2家族参与脂多糖诱导的肾小管上皮细胞凋亡  

Pyruvate kinase regulates apoptosis of renal tubular epithelial cells in LPS-induced acute kidney injury via Bcl-2 family

作　　者：赵延芳 胡锦秀 王荣[1] ZHAO Yanfang;HU Jinxiu;WANG Rong(Department of Nephrology,Shandong Provincial Hospital Affiliated to Shandong University,Jinan 250021,China)

机构地区：[1]山东大学附属省立医院肾内科,济南250021

出　　处：《免疫学杂志》2020年第5期390-396,共7页Immunological Journal

摘　　要：目的探讨丙酮酸激酶2(PKM2)在脂多糖(LPS)诱导人肾小管上皮细胞(HK-2)凋亡中的作用及分子机制。方法采用不同质量浓度的脂多糖作用于HK-2细胞,利用CCK-8实验检测细胞存活率;利用Real-time PCR、Western blot检测HK-2细胞中PKM2的mRNA及蛋白表达水平;利用Western blot检测Bcl-2家族关键蛋白B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)以及细胞色素C(Cytochrome C)的表达;利用Annexin-PE/7AAD双染法分析细胞凋亡水平;利用流式细胞术PI染色法检测细胞周期,并结合PKM2抑制剂,深入探讨PKM2在其中的作用及机制。结果脂多糖可诱导HK-2细胞损伤,使其活性降低、周期阻滞、凋亡增加,且可检测到Bcl-2家族关键蛋白的表达改变:Bcl-2降低、Bax增高,同时伴Cytochrome C增加。脂多糖可诱导PKM2表达增加,而使用PKM2抑制剂后,进一步增加了HK-2细胞凋亡以及细胞G1-S期阻滞,伴Bcl-2显著降低,Bax、Cytochrome C显著增加。结论脂多糖诱导HK-2细胞活力下降,周期阻滞,凋亡增加,PKM2可以通过靶向Bcl-2家族参与脂多糖诱导的HK-2细胞凋亡。To investigate the role of PKM2 in apoptosis of human renal tubular epithelial cells(HK-2) induced by lipopolysaccharide(LPS) and its molecular mechanism. Different concentrations of LPS were applied to HK-2 cells,Cell viability was measured by CCK8 assay;the expression of PKM2 were detected by Real-time PCR and Western blotting;Bcl-2, Bax and Cytochrome C were detected by Western blotting;apoptosis levels were analyzed by Annexin-PE/7 AAD double staining;cell cycle was detected by flow cytometry PI staining, and PKM2 inhibitors were combined to investigate the role and mechanism of PKM2. Data showed LPS could induce HK-2 cells injury,decrease cell activity, increase the cycle arrest and apoptosis, and changes of Bcl-2 family key proteins were detected: Bcl-2 decreased, Bax and Cytochrome C increased were. At the same time, LPS stimulation was accompanied by an increase in the expression of PKM2. Combined with PKM2 inhibitors, the results showed that the use of inhibitors further aggravated HK-2 cell apoptosis and G1-S phase arrest, accompanied by the significantly down-regulation of Bcl-2, and increase of Bax and Cytochrome C. LPS can induce HK-2 cells injury, decrease cell viability, increase the cycle arrest and apoptosis. Pyruvate kinase regulates apoptosis of renal tubular epithelial cells in LPS-induced acute kidney injury via Bcl-2 family.

关 键 词：丙酮酸激酶2 BCL-2家族 HK-2细胞 脂多糖 凋亡 

分 类 号：R392.12[医药卫生—免疫学]