摘要
目的探究CK1α表达与糖尿病心肌病细胞自噬、脂质堆积、细胞凋亡的关系。方法构建新生心肌细胞(NCM)脂毒性模型以及2型糖尿病雄性db/db小鼠模型分别进行体外研究以及体内研究,体外研究中将心肌细胞脂毒性模型分为腺病毒介导CK1α基因高表达组、阴性对照组、CRISPR/Cas9介导CK1α基因敲除组,体内研究的20只2型糖尿病雄性db/db小鼠模型分为CK1α过表达db/db组、注射对照组,并选取10只C57BL/Ks正常小鼠纳入正常小鼠组,对比各组的细胞自噬、脂质堆积、细胞凋亡指标差异。结果⑴Beclin-1蛋白主要在胞浆及细胞核中分布,阴性对照组的CK1α、Beclin-1蛋白、mRNA比3MA组高(P<0.05);⑵腺病毒介导CK1α基因高表达组的CK1α、Beclin-1蛋白、mRNA、Beclin-1蛋白绿色荧光强度最高,其次是阴性对照组、CRISPR/Cas9介导CK1α基因敲除组(P<0.05);⑶腺病毒介导CK1α基因高表达组的脂滴数量、二酰甘油及神经酰胺含量、细胞凋亡水平最低,其次是阴性对照组、CRISPR/Cas9介导CK1α基因敲除组(P<0.05);⑷CK1α过表达db/db组的LVIDd、LVIDs最低,FS、EF%水平更高,其次是正常小鼠组、注射对照组(P<0.05);⑸CK1α过表达db/db组具有最低的脂滴数量、二酰甘油及神经酰胺含量、细胞凋亡水平,其次是正常小鼠组、注射对照组(P<0.05);⑹CK1α过表达db/db组中CK1α、Beclin-1蛋白、mRNA表达水平最高,其次是正常小鼠组、注射对照组(P<0.05)。结论CK1α表达高表达会增强细胞自噬,CK1α表达可通过增强自噬能力途径改善糖尿病心肌病的脂质沉积、抑制细胞凋亡以及改善心功能的作用,可将CK1α与细胞自噬作为后续糖尿病心肌病治疗的新方向。
Objective To investigate the relationship between the expression of CK1αand autophagy,lipid accumulation and apoptosis in diabetic cardiomyopathy.Methods Neonatal cardiomyocyte(NCM)lipotoxicity model and type 2 diabetic male DB/DB mouse model were established for in vitro and in vivo study,respectively.In vitro study,cardiomyocyte lipotoxicity model was divided into adenovirus-mediated CK1αgene high expression group,negative control group,CRISPR/Cas9-mediated CK1αgene knockout group.In vivo,20 male db/db mice with type 2 diabetes were divided into CK1αoverexpression db/db group and injection control group,and 10 C57BL/Ks normal mice were included in the normal mice group,and the differences of autophagy,lipid accumulation and apoptosis indexes were compared among each group.Results(1)Beclin-1 protein was mainly distributed in cytoplasm and nucleus,and the levels of CK1α,Beclin-1 protein and mRNA in negative control group increased than those in 3MA group(P<0.05);(2)The green fluorescence intensity of CK1α,Beclin-1 protein,mRNA and Beclin-1 protein in adenovirus-mediated CK1αhigh expression group was the highest,followed by the negative control group,CRISPR/Cas9-mediated CK1αknockout group(P<0.05);(3)The number of lipid droplets,the contents of dicylglycerol and neuramide and the apoptosis level of adenovirus-mediated CK1αgene high expression group were the lowest,followed by the negative control group,CRISPR/Cas9-mediated CK1αgene knockout group(P<0.05);(4)The LVIDd and LVIDs of CK1αoverexpressing db/db group were the lowest,and FS and EF%increased in CK1αoverexpressing db/db group,followed by normal mice group and injection control group(P<0.05);(5)CK1αoverexpressed db/db group had the lowest number of lipid droplets,the contents of dicylglycerol and amide,and the level of apoptosis,followed by normal mice group,injection control group(P<0.05);(6)The expression levels of CK1α,Beclin-1 protein and mRNA in db/db overexpressed CK1αgroup were the highest,followed by normal mice group and injection con
作者
余华
谷翔
张军
周利伟
YU Hua;GU Xiang;ZHANG Jun(Department of Cardiology,Jiujiang University Affiliated Hospital,Jiangxi Jiujiang 332000)
出处
《江西医药》
CAS
2021年第12期2118-2123,共6页
Jiangxi Medical Journal
基金
江西省卫生健康委科技计划项目,编号20202047。
