高迁移率族蛋白1通过调控自噬因子P62调节心肌细胞纤维化  

HMGB1 induces myocardial fibrosis by regulating autophagy factor P62

作　　者：聂雅琴 乌日娜[2] 金慧[2] 张广平[2] 李永明[2] 智利 薛仕煊 娜日格乐 NIE Yaqin;WU Rina;JIN Hui;ZHANG Guangping;LI Yongming;ZHI Li;XUE Shixuan;NARI Gele(Baotou Medical College Center Clinical Medical College,Baotou 014040;Cardiovascular Department of Baotou Central Hospital,Baotou 014040,China)

机构地区：[1]包头医学院中心临床医学院,014040 [2]包头市中心医院心血管内科,014040

出　　处：《国际心血管病杂志》2024年第2期109-113,共5页International Journal of Cardiovascular Disease

基　　金：内蒙古医科大学科技百万工程联合项目[YKD2020KJBW(LH)065]。

摘　　要：目的:探究高迁移率族蛋白1(HMGB1)通过调控自噬活性对心肌纤维化的影响及作用机制。方法:将人心肌细胞AC16培养于DMEM培养基中,用不同浓度的HMGB1(0、4、20、100μg/L)干预AC16细胞6 h,免疫荧光染色检测心脏成纤维细胞活化的标志性蛋白α平滑肌肌动蛋白(α-SMA)的表达水平,Western blot法检测心肌自噬相关蛋白哺乳动物雷帕霉素靶蛋白(mTOR)、磷酸化mTOR(p-mTOR)、微管相关蛋白1轻链3(LC3)、磷酸肌醇-3-激酶3(PIK3C3)、可溶性及不可溶性P62、α-SMA、Ⅰ型胶原蛋白(CollagenⅠ)在AC16中的表达水平。免疫共沉淀检测α-SMA与P62之间的互相作用。结果:不同浓度的HMGB1刺激AC16细胞,AC16细胞中心肌纤维化标志蛋白CollagenⅠ与α-SMA的表达水平呈浓度依赖性升高,自噬蛋白PIK3C3、不可溶性P62的蛋白表达水平,p-mTOR与mTOR的比值,LC3-Ⅱ与LC3-Ⅰ的比值呈浓度依赖性升高,可溶性P62的蛋白表达水平呈浓度依赖性降低。各组间的差异均有统计学意义(P均<0.01)。免疫共沉淀实验显示P62与α-SMA存在相互作用。结论:外源性HMGB1可抑制AC16细胞中的自噬流,诱导AC16细胞纤维化。Objective:To explore the effect of high mobility group box protein 1(HMGB1)on myocardial fibrosis by regulating autophagy activity of human cardiomyocytes(AC16)and its possible mechanism.Methods:AC16 cells were cultured in DMEM medium and treated with different concentrations of HMGB1(0、4、20、100μg/L)for 6 h.The expression ofα-smooth muscle actin(α-SMA),a marker protein of cardiac fibroblast activation,was detected by immunofluorescence staining.Western blot was used to determine the expression levels of myocardial autophagy-related proteins mammalian target of rapamycin(mTOR),phosphorylated mTOR(p-mTOR),light chain 3(LC3),phosphoinositide-3-kinase 3(PIK3C3),soluble and insoluble p62,α-SMA,and collagenⅠ(CollagenⅠ)in AC16.The interaction betweenα-SMA and p62 was assessed by co-immunoprecipitation.Results:The expression levels of myocardial fibrosis marker protein(collagenⅠandα-SMA),autophagy protein PIK3C3,insoluble p62,P-mTOR/mTOR ratio,LC3-Ⅱ/LC3-Ⅰratio were increased in a dose-dependent manner.In contrast,the expression level of soluble p62 was decreased in a dose-dependent manner.The differences between groups were statistically significant(all P<0.01).There was an interaction between p62 andα-SMA.Conclusion:Exogenous HMGB1 inhibits autophagic flux in AC16 cells and induces myocardial fibrosis.

关 键 词：高迁移率族蛋白1 AC16细胞 自噬 心肌纤维化 

分 类 号：R542.2[医药卫生—心血管疾病]