自噬对于风湿性心脏病瓣膜间质细胞分化以及增殖能力的影响  

Effect of autophagy on the differentiation and proliferative capacity of interstitial cells in rheumatic heart disease valves

作　　者：贺洲 谢晓勇[1] 罗程[1] 黎玉贵 温大帅 朱晓莉 He Zhou;Xie Xiaoyong;Luo Cheng;Li Yugui;Wen Dashuai;Zhu Xiaoli(Department of Cardiothoracic Surgery,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院心胸外科,南宁530021

出　　处：《中华实验外科杂志》2023年第1期87-89,共3页Chinese Journal of Experimental Surgery

基　　金：广西自然科学基金面上项目(2019GXNSFAA185055)。

摘　　要：目的探讨自噬对于风湿性心脏病的瓣膜间质细胞(VICs)生物学特征改变以及增殖能力的影响。方法使用肝素诱导人主动脉瓣膜间质细胞(购自上海赛百慷生物技术股份有限公司)向肌成纤维细胞分化,而后使用自噬抑制剂LY294002(15μmol/L)进行干预,将细胞分为对照组、抑制剂组、诱导组、实验组。使用蛋白质印迹法(Western blot)检测各组细胞α-平滑肌肌动蛋白(α-SMA)、碱性磷酸酶(ALP)、Beclin1、微管相关蛋白轻链3-Ⅱ(LC3-Ⅱ)蛋白表达水平;采用细胞增殖及毒性检测试剂盒(CCK-8)分析自噬对4组细胞增殖的影响。结果蛋白印记结果显示对照组的ALP和α-SMA蛋白(0.827±0.123、0.716±0.141)显著低于诱导组(1.384±0.165、1.315±0.223),差异有统计学意义(t=4.669,P<0.01、t=3.929,P<0.05);对照组Beclin1和LC3-Ⅱ(0.807±0.182、0.392±0.150)显著低于诱导组(1.271±0.067、1.404±0.018),差异有统计学意义(t=4.131,P<0.05、t=11.535,P<0.01);诱导组的ALP和α-SMA蛋白(1.385±0.165、1.447±0.014)显著高于实验组(0.939±0.189、0.907±0.324),差异有统计学意义(t=2.882,P<0.05)。CCK-8细胞增殖实验结果显示各组VICs细胞的细胞活力差异有统计学意义(F=545.843,P<0.01,η2=0.986),对照组细胞活力显著高于诱导组,差异有统计学意义(t=29.753,P<0.01,d=1.101);实验组细胞活力也显著高于诱导组,差异有统计学意义(t=14.034,P<0.01,d=0.878)。结论自噬促进风湿性心脏病瓣膜间质细胞分化,同时抑制其细胞活性。Objective To investigate the effect of autophagy on the altered biological characteristics and proliferative capacity of valvular interstitial cells(VICs)in rheumatic heart disease.Methods Heparin was used to induce differentiation of human aortic VICs to myofibroblasts,while the cells were later intervened with the autophagy inhibitor LY294002(15μmol/L),and the cells were divided into control,inhibitor,induction,and experimental groups.The expression levels of α-smooth muscle actin(α-SMA),alkaline phosphatase(ALP),Beclin1,and microtubule-associated protein light chain 3-Ⅱ(LC3-Ⅱ)protein were detected in each group of cells using Western blotting.The effect of autophagy on the proliferation of cells in the four groups was analyzed using the CCK-8 assay.Results The protein blotting results showed that the expression levels of ALP andα-SMA proteins in the control group(0.827±0.123,0.716±0.141)were significantly lower than those in the induction group(1.384±0.165,1.315±0.223,t=4.669,P<0.01;t=3.929,P<0.05).The expression of Beclin1 and LC3-Ⅱ in the control group(0.807±0.182,0.392±0.150)was significantly lower than that in the induction group(1.271±0.067,1.404±0.018,t=4.131,P<0.05;t=11.535,P<0.01).The expression levels of ALP andα-SMA proteins in the induction group(1.385±0.165,1.447±0.014)were significantly higher than those in the experimental group(0.939±0.189,0.907±0.324,t=2.882,P<0.05).The results of CCK-8 assay showed that the viability of VICs in each group was significantly different and the difference was statistically significant(F=545.843,P<0.01,η2=0.986).The cell viability of the control group was significantly higher than that of the induction group(t=29.753,P<0.01,d=1.101).The cell viability of the experimental group was also significantly higher than that of the induction group(t=14.034,P<0.01,d=0.878).Conclusion Autophagy promotes differentiation of rheumatic disease heart valve mesenchymal cells and inhibits their cellular activity.

关 键 词：风湿性心脏病 瓣膜间质细胞 自噬 

分 类 号：R541.2[医药卫生—心血管疾病]