藜芦醇对CoCl2诱导H9C2心肌细胞损伤的保护作用研究

doi:10.3969/j.issn.10073205.2018.08.003

摘要/Abstract

摘要： ［摘要］〓
〖HTH〗目的〖HTSS〗〖KG*2〗探讨白藜芦醇（Resveratrol,RES）对二氯化钴（CoCl2）诱导H9C2心肌细胞损伤保护作用的机制。
〖HTH〗方法〖HTSS〗〖KG*2〗500 μmol/L CoCl2处理H9C2心肌细胞24 h以建立H9C2心肌细胞缺氧损伤模型。实验分组：空白对照组（control）；模型损伤组（0.1% DMSO培养液预处理H9C2细胞24 h后，500 μmol/L CoCl2继续处理H9C2细胞24 h）；低剂量RES组（12.5 μmol/L RES预处理H9C2细胞24 h后，500 μmol/L CoCl2继续处理H9C2细胞24 h）；中剂量RES组（25 μmol/L RES预处理H9C2细胞24 h后，500 μmol/L CoCl2继续处理H9C2细胞24 h）；高剂量RES组（50 μmol/L RES预处理H9C2细胞24 h后，500 μmol/L CoCl2继续处理H9C2细胞24 h）。采用MTT法评价H9C2心肌细胞活力。Hoeschst 33342染色观察H9C2细胞核形态的变化。采用流式细胞技术检测H9C2细胞线粒体膜电位的变化、细胞内活性氧的产生及细胞凋亡的情况。Western blot 检测H9C2细胞中凋亡相关蛋白的变化。
〖HTH〗结果〖HTSS〗〖KG*2〗MTT结果显示，RES浓度依赖性抑制CoCl2诱导的H9C2细胞活力的降低；Hoeschst 33342染色观察显示，RES浓度依赖性抑制CoCl2引起的H9C2细胞凋亡形态的出现；流式细胞术显示，RES浓度依赖地抑制CoCl2诱导的H9C2细胞内活性氧的产生、线粒体膜电位的降低、细胞凋亡的增加；Western blot检测结果显示，RES浓度依赖性抑制CoCl2引起的H9C2细胞中促凋亡蛋白（Bax、cleavedcaspase3、cleavedcaspase9）的增加和抑凋亡蛋白（Bcl2）的降低。
〖HTH〗结论〖HTSS〗〖KG*2〗RES对CoCl2引起的H9C2心肌细胞损伤具有保护作用，其机制可能是通过抑制CoCl2引起的活性氧增加、线粒体膜电位降低进而阻碍H9C2细胞线粒体凋亡的发生而实现的。

关键词: 肌细胞, 心脏；白藜芦醇；细胞凋亡

Abstract: ［Abstract］ Objective〖HTSS〗〓To study the protective mechanism of resveratrol(RES) on H9C2 cells injury induced by two cobalt chloride(CoCl2).
〖HTH〗〖WTHZ〗Methods〖HTSS〗〓CoCl2 （500 μmol/L） was used to treat H9C2 cardiac muscle cell for 24 h in order to establish H9C2 myocardial cell hypoxia injury model. The experiment was divided into five groups: blank control group(control); model injury group(H9C2 cells were pretreated with 0.1% DMSO culture medium for 24h, and then they were continued to treat by 500μmol/L CoCl2 for 24h); low dose RES group(H9C2 cells were pretreated with 12.5μmol/L RES for 24h, and then they were continued to treat by 500μmol/L CoCl2 for 24h); medium dose RES group(H9C2 cells were pretreated with 25μmol/L RES for 24h, and then they were continued to treat by 500μmol/L CoCl2 for 24h); high dose RES group(H9C2 cells were pretreated with 50μmol/L RES for 24h, and then they were continued to treat by 500μmol/L CoCl2 for 24h). MTT method was used to evaluate the viability of H9C2 myocardial cell. Morphological changes of H9C2 nucleus were observed by Hoeschst 33342 staining. The changes of mitochondrial membrane potential, intracellular reactive oxygen species(ROS) production and cell apoptosis were detected by flow cytometry in H9C2 cells. Western Blot was used to investigate the changes of apoptosis related proteins in H9C2 cells.
〖HTH〗〖WTHZ〗Results〖HTSS〗〓MTT results showed that RES concentration dependently inhibited the decrease of H9C2 cell viability induced by CoCl2. Hoeschst 33342 staining showed that RES concentration dependently inhibited the appearance of H9C2 cells apoptosis induced by CoCl2. Flow cytometry revealed that RES concentration dependently inhibited ROS production, decreased mitochondrial membrane potential, and increased apoptosis in H9C2 cells induced by CoCl2. Western blot showed that RES concentration dependently inhibited the increase of proapoptotic proteins(Bax, cleavedcaspase3, cleavedcaspase9) and decreased expression of apoptotic protein(Bcl2) in H9C2 cells induced by CoCl2.
〖HTH〗〖WTHZ〗Conclusion〖HTSS〗〓RES has the protective effect on H9C2 myocardial cell injury induced by CoCl2, the mechanism may be related with inhibiting the increase of ROS induced by CoCl2 and decreasing the mitochondrial membrane potential, thus hindering the occurrence of mitochondrial apoptosis in H9C2 cells.

Key words: myocytes, cardica, Resveratrol, apoptosis

杜〓峥，王〓巍，路〓军，苏海明*. 藜芦醇对CoCl2诱导H9C2心肌细胞损伤的保护作用研究[J]. 河北医科大学学报, doi: 10.3969/j.issn.10073205.2018.08.003.

DU Zheng, WANG Wei, LU Jun, SU Haiming*.

Study on the protective effect of Resveratrol on H9C2 myocardial cell injury induced by CoCl2

[J]. Journal of Hebei Medical University, doi: 10.3969/j.issn.10073205.2018.08.003.

[1]	刘钰, 王星, 崔红占, 孙涌泉, 步纪强, 陈子英. 利拉鲁肽通过circ-sirt1/NF-κB信号通路抑制血管平滑肌细胞炎症的机制研究 [J]. 河北医科大学学报, 2022, 43(9): 996-1001.
[2]	闻松, 徐正宇(综述), 蔡子怡, 刘巍(审校). 慢性肾病血管钙化的发病机制研究进展[J]. 河北医科大学学报, 2022, 43(2): 233-239.
[3]	卢玉润, 唐宇帆. 黄连素通过miR-29b保护缺氧复氧诱导H9c2细胞损伤中的作用及其机制[J]. 河北医科大学学报, 2021, 42(3): 256-261,267.
[4]	张亚楠1，陈〓炜1，张金平1，齐莉莉2，张〓雷1，赵春芳1*. 过表达GATA4的骨髓间充质干细胞向心肌细胞分化的研究[J]. 河北医科大学学报, 2019, 40(8): 877-881,901.
[5]	贡〓悦，王齐松，王凡轲，孟泽琪，刘晓宁，聂〓磊* . 跨膜蛋白ESDN的表达与血管内膜増生的初步研究#br#[J]. 河北医科大学学报, 2019, 40(7): 753-757,798.
[6]	庞雅湘1，聂启昊2，刘晓宁3，王〓帅3，冯〓琦3，牛洪琳4*. 3种细胞裂解液提取总蛋白在Western blot中的效果分析[J]. 河北医科大学学报, 2019, 40(3): 263-267.
[7]	张曼莉1，张曼娜2，佟〓飞1，温进坤3. KLF5与NFκB p50相互作用促进高糖诱导的血管平滑肌细胞炎症[J]. 河北医科大学学报, 2018, 38(12): 1371-1375,1379.
[8]	黄海霞;侯东燕;辛方;任杰;刘萍;王伟. 肌动蛋白对结肠平滑肌大电导钙激活钾通道机械敏感性的影响[J]. , 2015, 36(11): 1245-1245.
[9]	陈鹏;赵丽丽;聂茜;李亮;吴珺;韩梅. 贴块法培养大鼠主动脉血管平滑肌细胞条件的优化[J]. , 2014, 35(10): 1220-1220.
[10]	李伟聪;杨晶;魏燕;张翼;朱晓光;张黎. 慢性间歇性低压低氧对大鼠心室肌细胞瞬时外向和稳态外向钾通道电流的影响[J]. , 2013, 34(4): 373-373.
[11]	王梅;李拥军;刘素云;张辉;杨蓉;苗成龙. 胰岛素抵抗心肌细胞模型的建立与评价[J]. , 2012, 33(9): 993-993.
[12]	斯琴高娃;魏燕;宋胜利;周京京;杨晶;张翼. 慢性间歇性低压低氧对大鼠心肌细胞内向整流钾通道的影响[J]. , 2012, 33(7): 745-745.