姜黄素对布比卡因诱导的心肌细胞毒性的作用及机制研究  

The role and mechanism of curcumin in bupivacaine-induced cardiotoxicity in cardiomyocytes

作　　者：司海超[1] 付莉萍[2] 刘展[1] 高毅[1] 司小萌[1] 马婕妤[3] 

机构地区：[1]南阳市中心医院麻醉科,河南南阳473000 [2]南阳市中心医院眼科 [3]郑州市中心医院麻醉科

出　　处：《毒理学杂志》2017年第6期451-455,共5页Journal of Toxicology

摘　　要：目的探究姜黄素对布比卡因诱导的心肌细胞毒性作用及机制。方法首先用WST-1法检测不同浓度姜黄素(0、25、50、75和100μmol/L)对H9C2心肌细胞增殖的影响,确定姜黄素的最适浓度。然后用最适浓度的姜黄素处理H9C2细胞24 h后,加入1 mmol/L布比卡因刺激24 h,WST-1检测细胞活力,流式细胞术检测细胞凋亡,Western blotting方法检测细胞中细胞色素C(Cyt C)、活化的Caspase-3、Bax、Bcl-2、p-Akt和p-GSK-3β的表达水平。最后,用GSK-3β抑制剂TWS119处理H9C2细胞,检测GSK-3β通路抑制后姜黄素对布比卡因诱导的细胞增殖和凋亡相关蛋白表达的影响。结果 75μmol/L开始抑制细胞增殖,因此姜黄素浓度的选择为50μmol/L。姜黄素能够缓解布比卡因对心肌细胞增殖的抑制,减少细胞凋亡,降低Cyt C和Caspase-3的表达及Bax/Bcl-2值,增加p-Akt和p-GSK-3β的表达。TWS119能减弱姜黄素对布比卡因诱导的心肌细胞增殖、Cyt C和活化的Caspase-3表达及Bax/Bcl-2值的影响。结论姜黄素能缓解布比卡因诱导的心肌细胞毒性,这与其激活Akt/GSK-3β信号通路、抑制线粒体凋亡通路相关。Objective To investigate the role and molecular mechanism of curcumin in bupivacaine-induced cardiotoxicity in cardiomyocytes. Methods The effects of different concentrations of curcumin （0, 25, 50, 75, 100 μmol/L） on the proliferation of H9C2 cardiomyocytes were detected by WST-1 method to confirm the proper concentration of curcumin in the following study. Subsequently, H9C2 cells were treated with indicated concentration of curcumin for 24 h, and then stimulated with 1 mmol/L bupivacaine for 24 h. Cell viability was determined by WST-1 assay, and cell apoptosis was assayed by flow cytometry. The levels of cytochrome C （CytC） , active Caspase-3, Bax, Bc1-2, p-Akt, and p-GSK-3β were analyzed by Western blotting. Additionally, H9C2 cells were treated with TWSll9, GSK-3β inhibitor, to assay the effects of curcumin on bupivacaine-induced cell proliferation and apoptosis related protein. Results 25 μmol/L and 50 μmol/L curcumin had no effect on cell proliferation, which could be inhibited from 75 μmol/L; hence, 50 μmol/L was used in the following study. Curcumin could attenuate bupivacaine-induced the inhibition of cell proliferation, the increase of cell apoptosis, the higher levels of CytC and active Caspase-3, and the enhanced ratio of Bax/Bcl-2, the reduction of p-Akt and p-GSK-3β levels. However, TWS119 could reduce the effects of curcumin on bupivacaine-induced cell proliferation and the expression levels of CytC and active Caspase-3, and the ratio of Bax/Bcl-2. Conclnsion Curcumin could attenuate bupivacaine-induced cardiotoxicity in cardiomyocytes, which is related with the activation of Akt/GSK-3β signaling and the inhibition of mitochondrial apoptotic pathway.

关 键 词：姜黄素 布比卡因 心肌细胞 毒性 Akt/GSK-3β信号通路 

分 类 号：R971.2[医药卫生—药品]