敲低Spectrin β Ⅱ加重棕榈酸诱导的心肌细胞损伤  

Spectrin β Ⅱ knockdown aggravates cardiomyocyte injury induced by palmitic acid

作　　者：杨容金 邢萍萍 张顺 张晓萌 张飞宇 王姗 陶凌 YANG Rong-jin;XING Ping-ping;ZHANG Shun;ZHANG Xiao-meng;ZHANG Fei-yu;WANG Shan;TAOLing(Department of Cardiology,Xijing Hospital,Air Force Medical University,Xi’an 710032,Shaanxi,China)

机构地区：[1]空军军医大学西京医院心内科,陕西西安710032

出　　处：《心脏杂志》2022年第2期125-129,共5页Chinese Heart Journal

基　　金：国家自然科学基金项目资助(82070867,81730011)。

摘　　要：目的研究血影蛋白β Ⅱ(Spectrin β Ⅱ)在棕榈酸诱导的小鼠原代心肌细胞DNA损伤和细胞凋亡中的作用。方法(1)Western blot检测棕榈酸处理后小鼠原代心肌细胞Spectrin β Ⅱ表达量变化。(2)分别用Ad-shscramble(对照组)和Ad-sh-Spectrin β Ⅱ(Spectrin β Ⅱ干涉组)的腺病毒感染小鼠原代心肌细胞,荧光显微镜下观察病毒的感染效率。利用Western blot和qPCR检测Spectrin β Ⅱ的干涉效率。(3)Western blot检测棕榈酸处理后Ad-sh-scramble组和Spectrin β Ⅱ干涉组小鼠原代心肌细胞DNA损伤指标γ-H2AX及凋亡蛋白剪切型半胱天冬酶3(cleaved-caspase 3)的表达,采用流式细胞术检测各组细胞凋亡水平。结果(1)给予棕榈酸处理的小鼠原代心肌细胞Spectrin β Ⅱ蛋白水平显著降低(P<0.05)。(2)与Ad-sh-scramble组相比,Ad-sh-Spectrin β Ⅱ组Spectrin β Ⅱ的蛋白和mRNA水平均显著降低(P<0.05)(3)给予棕榈酸处理后,与Ad-sh-scramble组相比,Ad-sh-Spectrin β Ⅱ组的γ-H2AX2和cleaved-caspase 3蛋白表达显著升高(P<0.05),并且细胞凋亡率显著升高(P<0.05)。结论敲低Spectrin β Ⅱ加重棕榈酸诱导的小鼠原代心肌细胞DNA损伤和细胞凋亡。AIM To investigate the effects of Spectrin β Ⅱ on cardiomyocyte DNA damage and apoptosis induced by palmitic acid(PA).METHODS Western blot was used to detect the protein levels of Spectrin β Ⅱ in mouse primary cardiomyocytes(MCMs)treated with PA.MCMs were infected with adenovirus expressing independent hairpin shRNA sequences for Spectrin β Ⅱ (Ad-sh-Spectrin β Ⅱ )or control sh-scramble sequence(Ad-sh-scramble).The infection efficiency of the adenovirus was examined by fluorescence microscope and confirmed by Western blot and qPCR.The protein levels ofγ-H2AX,an indicator of DNA damage,and apoptotic protein cleaved-caspase 3 were detected by Western blot.Flow cytometry was used to analyze the apoptotic rate.RESULTS The protein levels of Spectrin β Ⅱ were significantly decreased in MCMs treated with PA(P<0.05).Compared with those in the Ad-sh-scramble group,the protein and mRNA levels of Spectrin β Ⅱ in the Ad-sh-Spectrin β Ⅱ group were significantly decreased(P<0.05).Compared with those in the Ad-sh-scramble group,the protein levels ofγ-H2AX and cleaved-caspase3 were significantly increased(P<0.05)and the apoptotic rate was significantly increased(P<0.05)in the Ad-sh-Spectrin β Ⅱ group after treated with PA.CONCLUSION Spectrin β Ⅱ knockdown aggravates cardiomyocyte DNA damage and apoptosis induced by palmitic acid.

关 键 词：血影蛋白βⅡ 心肌细胞 棕榈酸 DNA损伤 细胞凋亡 

分 类 号：R541.9[医药卫生—心血管疾病]