木犀草素调控内质网应激-线粒体凋亡通路在脂毒性心肌损伤中的作用  

The Role of Luteolin in Regulating the Endoplasmic Reticulum Stress-mitochondrial Apoptosis Pathway in Lipotoxic Myocardial Injury

作　　者：项云 梁潇 鲍翠玉[2] 刘涛 XIANG Yun;BAO Cui-yu;LIU Tao(School of Pharmacy,Xianning Medical College,Hubei University of Science and Technology,Xianning Hubei 437100,China)

机构地区：[1]湖北科技学院医学部药学院,湖北咸宁437100 [2]湖北科技学院糖尿病心脑血管病变湖北省重点实验室 [3]湖北科技学院附属第二医院

出　　处：《湖北科技学院学报（医学版）》2023年第3期185-189,195,F0002,共7页Journal of Hubei University of Science and Technology(Medical Sciences)

基　　金：国家自然科学基金项目(51703055);湖北省自然科学基金项目(2020CFB838);咸宁市科技计划项目(2018-49,2019kj03);湖北科技学院校内培育科研项目(2021-22X16);湖北科技学院五官医学院专项科研基金项(2020WG14);湖北科技学院校内科研发展项目附属第二医院专项(2021LCZ001)。

摘　　要：目的探讨木犀草素(LU)对H9c2心肌细胞的保护作用及与内质网应激-线粒体凋亡通路的关系。方法使用棕榈酸(PA)建立H9c2心肌细胞脂毒性损伤的模型,将实验组分为正常组、模型组及木犀草素组(15μmol/L)。采用CCK8法测定体外培养的细胞增殖水平,利用试剂盒检测细胞内超氧化物歧化酶(SOD)、丙二醛(MDA)水平,使用DCFH-DA检测细胞内活性氧(ROS)含量,JC-1荧光探针检测线粒体膜电位变化,Annexin V-FITC/PI测定细胞凋亡程度,Westren blot法检测细胞在内质网应激-线粒体凋亡通路中相关基因的表达。结果用PA(0.2mmol/L)直接刺激24h,可明显降低H9c2心肌细胞增殖率,增加细胞氧化应激水平,降低线粒体膜电位,使p-PERK、p-IRE1、GRP78、CHOP、Bax、caspase-3蛋白表达显著升高,Bcl-2蛋白表达降低,但使用木犀草素处理后,明显逆转了以上结果,降低了PA诱导的H9c2心肌细胞氧化应激水平,减轻细胞凋亡程度,逆转了在PA影响下细胞内内质网应激-线粒体凋亡通路相关蛋白的表达。结论木犀草素可减轻PA诱导的脂毒性心肌损伤,且这一过程可能与内质网应激-线粒体凋亡通路有关。Objective To investigate the protective effect of Luteolin(LU)on H9c2 cardiomyocytes and its relationship with the endoplasmic reticulum stress-mitochondrial apoptosis pathway.Methods Palmitic acid(PA)was used to establish the model of H9c2 cardiomyocyte lipotoxic injury.The experimental groups were divided into the normal group,model group,and LU group(15μmol/L).The proliferation level of cultured cells was measured by the CCK8 method,the intracellular superoxide dismutase(SOD)and malondialdehyde(MDA)levels were detected by kits,the intracellular reactive oxygen species(ROS)content was detected by DCFH-DA,and the changes of mitochondrial membrane potential were detected by JC-1 fluorescent probe.Annexin V-FITC/PI was used to determine the degree of apoptosis,and western blot was used to detect the expression of related genes in the endoplasmic reticulum stress-mitochondrial apoptosis pathway.Results Direct stimulation with PA(0.2 mmol/L)for 24 h significantly decreased the proliferation rate of H9c2 cardiomyocytes,increased the oxidative stress level of cells,decreased the mitochondrial membrane potential,significantly increased the protein expressions of p-PERK,p-IRE1,GRP78,CHOP,Bax,and caspase-3,and decreased the expression of Bcl-2.However,the above protein expressions were significantly reversed after treatment with LU,which meanwhile reduced the level of PA-induced oxidative stress in H9c2 cardiomyocytes,alleviated the degree of apoptosis,and reversed the expression of proteins related to the endoplasmic reticulum stress-mitochondrial apoptosis pathway under the influence of PA.Conclusion These results suggest that LU can alleviate PA-induced lipotoxic myocardial injury,and this process may be related to the endoplasmic reticulum stress-mitochondrial apoptosis pathway.

关 键 词：心肌细胞 高脂 木犀草素 内质网应激 凋亡 

分 类 号：R961[医药卫生—药理学]