中介素预处理对NRK-52E细胞缺氧/复氧损伤的保护作用及其机制  

Protective effect of intermedin preconditioning against hypoxic/reoxygenation injury in NRK⁃52E cells and its mechanism

作　　者：冯殿卿 薛福平[2] 张媛媛 王艳红 FENG Dianqing;XUE Fuping;ZHANG Yuanyuan;WANG Yanhong(Taiyuan Center for Disease Control and Prevention,Tai-yuan 030012,China;Department of Nephrology,Armed Police Shanxi Corps Hospital;School of Basic Medical Sciences,Shanxi Medi-cal University)

机构地区：[1]太原市疾病预防控制中心,太原030012 [2]武警山西总队医院肾病科 [3]山西医科大学基础医学院

出　　处：《山西医科大学学报》2024年第10期1308-1313,共6页Journal of Shanxi Medical University

基　　金：山西省基础研究计划面上项目(202203021221188,201901D111187)。

摘　　要：目的 探讨中介素(intermedin,IMD)在缺氧/复氧(hypoxia/reoxygenation,H/R)条件下对大鼠近端肾小管上皮细胞(NRK-52E)损伤的保护作用及其机制。方法 体外培养NRK-52E细胞,随机分为空白对照(NC)组、氯化钴(CoCl2)组和IMD+CoCl2组;采用1.5×10^(-4) mol/L CoCl2处理24 h建立NRK-52E细胞H/R模型,IMD+CoCl2组采用IMD 20 ng/mL预处理4 h,保持IMD浓度(20 ng/mL)用1.5×10^(-4) mol/L CoCl2处理24 h。流式细胞术检测细胞凋亡率,DCFH-DA探针法检测细胞内活性氧(ROS)含量,JC-1探针法检测线粒体膜电位(MMP),real time-PCR法检测凋亡相关蛋白B淋巴细胞瘤-2(Bcl-2)、Caspase-3,以及氧化应激蛋白血红素加氧酶-1(HO-1)和内皮型一氧化氮合酶(eNOS)mRNA表达水平,Western blot法检测Bcl-2、Caspase-3、HO-1和eNOS蛋白表达。结果 与NC组相比,CoCl2组细胞凋亡率增加(P<0.01);ROS生成增加,MMP下降,同时Caspase-3表达增加(P<0.01),Bcl-2表达降低(P<0.01);与CoCl2组相比,IMD+CoCl2组细胞凋亡率显著下降(P<0.01),ROS生成显著减少而MMP显著增加,此外Caspase-3 mRNA及蛋白表达均下调(P<0.01),Bcl-2及eNOS、HO-1表达显著增加(P<0.05)。结论IMD能够改善NRK-52E细胞H/R诱导的损伤,其可能的机制是通过抑制H/R诱导的细胞凋亡和氧化应激,上调Bcl-2及eNOS、HO-1表达。Objective To explore the protective effect of intermedin(IMD)against rat renal tubular epithelial cell(NRK‐52E)injury induced by hypoxia/reoxygenation(H/R).Methods The NRK‐52E cells were randomly divided into normal control(NC)group,CoCl2 group and IMD+CoCl2 group.H/R injury models were induced by CoCl2(1.5×10^(-4) mol/L)for 24 h.The cells in IMD+CoCl2 group were incubated with IMD 20 ng/mL for 4 h,and then cultured with 20 ng/mL IMD and 1.5×10^(-4) mol/L CoCl2 for 24 h.Flow cytometry was used to access the apoptosis rate of cells,reactive oxygen species(ROS)generation was detected by DCFH‐DA probe,mitochon‐drial membrane potential(MMP)was detected by JC‐1 probe.Real time PCR was employed to detect the mRNA expression levels of Bcl‐2,Caspase‐3,HO‐1 and eNOS.Western blot was employed to detect the protein expression levels of Bcl‐2,Caspase‐3,HO‐1 and eNOS.Results Compared with NC group,the cell apoptosis rate was significantly increased in CoCl2 group(P<0.01),the intracellu‐lar ROS level was significantly increased and MMP was significantly decreased,the mRNA and protein expression levels of Caspase‐3 were significantly increased(P<0.01),and the protein expression of Bcl‐2 was significantly decreased(P<0.05).Compared with CoCl2 group,the cell apoptosis rate was significantly decreased in IMD+CoCl2 group(P<0.01),ROS level was decreased while MMP was increased,the mRNA and protein expressions of Caspase‐3 were significantly decreased(P<0.05),and the mRNA and protein expres‐sion levels of Bcl‐2,HO‐1 and eNOS were significantly increased(P<0.05).Conclusion IMD could ameliorate the hypoxia/reoxy‐genation‐induced injury in NRK‐52E cells by inhibiting cell apoptosis and oxidative stress,and increasing the expression levels of Bcl‐2,eNOS and HO‐1.

关 键 词：中介素 缺氧/复氧 NRK-52E细胞 细胞凋亡 氧化应激 线粒体膜电位 

分 类 号：R363[医药卫生—病理学]