布托啡诺通过调控miR-665表达对脂多糖致心肌细胞炎症反应及细胞凋亡的影响  

The Effect of Butorphanol on the Inflammatory Response and Apoptosis of Cardiomyocytes Induced by LPS by Regulating the Expression of miR-665

作　　者：魏利娟[1] 马亚飞[1] 陈小莉[1] 郭仲辉[1] 王鹏华 WEI Lijuan;MA Yafei;CHEN Xiaoli;GUO Zhonghui;WANG Penghua(School of Clinical Medicine,Henan University of Science and Technology,First Affiliated Hospital of Henan University of Science and Technology,Luoyang 471000,Henan,China)

机构地区：[1]河南科技大学临床医学院,河南科技大学第一附属医院,河南洛阳471000

出　　处：《中西医结合心脑血管病杂志》2022年第2期240-245,共6页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：河南省医学科技攻关计划项目(No.201602167);河南省科技计划项目(No.172102310686)。

摘　　要：目的探讨布托啡诺对脂多糖(LPS)致心肌细胞炎症反应、细胞凋亡的影响及其对微小RNA-665(miR-665)的调控作用。方法LPS处理心肌细胞建立细胞损伤模型,使用不同剂量的布托啡诺处理细胞,将anti-miR-NC、anti-miR-665分别转染至LPS诱导的心肌细胞,将miR-NC、miR-665 mimics分别转染至LPS诱导的心肌细胞后加入布托啡诺处理;采用酶联免疫吸附实验(ELISA)检测肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平;采用流式细胞术检测细胞凋亡率;采用实时荧光定量PCR(qRT-PCR)检测miR-665的表达量;蛋白免疫印迹法(Western Blot)检测Bcl-2、Bax蛋白表达量。结果LPS处理后可提高心肌细胞中TNF-α、IL-6水平、细胞凋亡率、Bax蛋白水平及miR-665的表达水平(P<0.05),降低Bcl-2蛋白水平(P<0.05);与LPS组比较,布托啡诺不同剂量组可明显降低TNF-α、IL-6水平、细胞凋亡率、Bax蛋白水平及miR-665的表达水平(P<0.05),提高Bcl-2蛋白水平(P<0.05);转染anti-miR-665对LPS诱导的心肌细胞炎症反应及凋亡的作用与布托啡诺相似;布托啡诺与转染miR-665 mimics共同处理后可明显提高TNF-α、IL-6水平、细胞凋亡率及Bax蛋白水平(P<0.001),降低Bcl-2蛋白水平(P<0.001)。结论布托啡诺可通过下调miR-665的表达而抑制LPS诱导的心肌细胞炎症反应及细胞凋亡,从而减轻心肌细胞损伤。Objective To explore the effect of butorphanol on the inflammatory response and apoptosis of cardiomyocytes induced by lipopolysaccharide(LPS),and its regulatory effect on miR-665.Methods Cardiomyocytes were treated with LPS to establish cell damage models,and the cells were treated with different doses of butorphanol.The anti-miR-NC and anti-miR-665 were transfected into LPS-induced cardiomyocytes.The miR-NC and miR-665 mimics were transfected into LPS-induced cardiomyocytes and then treated with butorphanol.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6).Flow cytometry was used to detect the apoptosis rate.Real-time fluorescent quantitative PCR(qRT-PCR)was used to detect the expression of miR-665.Western Blot method was used to detect the expression of Bcl-2 and Bax protein.Results LPS treatment could increase the levels of TNF-α,IL-6,apoptosis rate,the protein level of Bax,and the expression level of miR-665 in cardiomyocytes(P<0.05),and reduce the protein level of Bcl-2(P<0.05).Compared with the LPS group,different doses of butorphanol could significantly reduce the level of TNF-α,IL-6,apoptosis rate,the protein level of Bax,and the expression level of miR-665(P<0.05),and increase the protein level of Bcl-2(P<0.05).The effect of transfection of anti-miR-665 on the inflammatory response and apoptosis of cardiomyocytes induced by LPS was similar to that of butorphanol.Butorphanol and transfected miR-665 mimics could significantly increase the levels of TNF-α,IL-6,apoptosis rate,and the protein level of Bax(P<0.001),and reduce the protein level of Bcl-2(P<0.001).Conclusion Butorphanol could inhibit inflammatory response and apoptosis of cardiomyocytes induced by LPS through down-regulating the expression of miR-665,thus reducing cardiomyocyte damage.

关 键 词：心肌细胞 炎症反应 细胞凋亡 布托啡诺 微小RNA-665 脂多糖 实验研究 

分 类 号：R73[医药卫生—肿瘤]