原花青素B_(2)对LPS诱导的心肌细胞损伤的保护作用及机制  

Protective effect and mechanism of procyanidin B_(2) on LPS-induced cardiomyocyte injury

作　　者：李静 杨琴[1] 张雪峰[1] 杨吉明 龚芳 LI Jing;YANG Qin;ZHANG Xuefeng;YANG Jiming;GONG Fang(Department of Pharmacy,Jingzhou Second People's Hospital,Jingzhou,Hubei 434000,China;Department of Cardiology,Xiangyang Central Hospital&Affiliated Hospital of Hubei University of Arts and Science,Xiangyang,Hubei 441021,China)

机构地区：[1]荆州市第二人民医院药剂科,湖北省荆州市434000 [2]湖北文理学院附属医院襄阳市中心医院心内科,湖北省襄阳市441021

出　　处：《中国动脉硬化杂志》2021年第12期1028-1032,共5页Chinese Journal of Arteriosclerosis

基　　金：湖北省自然科学基金项目(WJ2015Q037)。

摘　　要：目的探讨原花青素B_(2)(PCB_(2))对LPS诱导的心肌细胞损伤的保护作用及机制。方法正常培养心肌细胞H9c2,用LPS诱导H9c2细胞建立细胞损伤模型,分别用6.25、12.5、25.0μmol/L的PCB_(2)处理模型细胞,25.0μmol/L的PCB_(2)处理模型细胞后加入核因子κB(NF-κB)信号通路抑制剂PDTC处理。采用MTT法检测细胞存活率;流式细胞术检测细胞凋亡率;酶联免疫吸附法(ELISA)检测细胞肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)和白细胞介素6(IL-6)的水平;丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)试剂盒分别检测MDA含量和SOD、GSH-Px活性;Westem blot检测细胞中NF-κB、IκB-α蛋白表达。结果LPS组细胞存活率较对照组显著降低(P<0.05),而PCB_(2)显著升高细胞存活率(P<0.05)。LPS组细胞凋亡率较对照组显著升高(P<0.05),而PCB_(2)显著降低LPS处理的细胞凋亡率(P<0.05)。LPS组细胞TNF-α、IL-1β、IL-6水平较对照组显著升高(P<0.05),而PCB_(2)显著降低LPS处理细胞TNF-α、IL-1β、IL-6水平(P<0.05)。与对照组比较,LPS组细胞MDA含量显著升高,SOD、GSH-Px活性显著降低(P<0.05);PCB_(2)显著降低LPS处理的细胞MDA含量,显著升高SOD、GSH-Px活性(P<0.05)。与对照组比较,LPS组细胞NF-κB蛋白表达显著升高,IκB-α蛋白表达显著降低(P<0.05);与LPS组比较,PCB_(2)显著降低细胞NF-κB蛋白表达,显著升高IκB-α蛋白表达(P<0.05)。与LPS+PCB_(2)组相比,LPS+PCB_(2)+PDTC能显著降低细胞凋亡率和TNF-α、IL-1β、IL-6、MDA含量,显著升高SOD、GSH-Px活性。结论PCB_(2)降低LPS诱导的心肌细胞凋亡率、炎症水平和氧化应激,提高细胞存活率,这可能与抑制NF-κB信号通路的活化有关。Aim To investigate the protective effect and mechanism of procyanidin B_(2)(PCB_(2))on lipopolysac-charide(LPS)-induced cardiomyocyte injury.Methods Cardiomyocytes H9c2 were cultured normally,and induced by LPS to establish cell damage models.The model cells were treated with PCB_(2)of 6.25,12.5 and 25.0μmol/L,25.0μmol/L PCB_(2)treatment model was then treated with NF-κB signaling pathway inhibitor PDTC treatment.Tetrazolium salt colorimetry(MTT)was used to detect cell survival;flow cytometry was used to detect cell apoptosis;enzyme-linked immu-nosorbent assay(ELISA)was used to detect cell tumor necrosis factor alpha(TNF-α),interleukin 1β(IL-1β)and inter-leukin 6(IL-6)levels;malondialdehyde(MDA),superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)kits were used to detect MDA content and activity of SOD and GSH-Px,respectively;Western blot was used to detect the expression of nuclear factor-κB(NF-κB)and IκB-αin cells.Results Compared with control group,the cell survival rate of the LPS group was significantly reduced(P<0.05);compared with LPS group,PCB_(2)significantly increased the cell survival rate(P<0.05).Compared with control group,the apoptosis rate was significantly increased in LPS group(P<0.05);compared with LPS group,PCB_(2)significantly reduced the apoptosis rate(P<0.05).Compared with control group,the levels of TNF-α,IL-1β,and were significantly increased IL-6 in LPS group(P<0.05);compared with LPS group,PCB_(2)significantly reduced the levels of TNF-α,IL-1β,and IL-6 in the cells(P<0.05).Compared with control group,the cell MDA content in LPS group was significantly increased,and the SOD and GSH-Px activities were significant-ly reduced(P<0.05);compared with LPS group,PCB_(2)significantly reduced the cellular MDA content and significantly increased the SOD and GSH-Px activities(P<0.05).Compared with control group,the cell NF-κB protein expression in LPS group was significantly increased,and the IκB-αprotein expression was significantly decreased(P<0.05);compared with LPS group,P

关 键 词：原花青素B_(2) LPS诱导 心肌细胞 细胞凋亡 

分 类 号：R363[医药卫生—病理学] R5[医药卫生—基础医学]