β-链蛋白基因沉默对高糖诱导血管内皮细胞钙化的抑制作用  

Inhibitory effect of β-catenin gene silencing on high glucose-induced endothelium calcification

作　　者：倪英群[1] 李居一[1] 施慧 牛云飞[1] 方朝晖[1] NI Ying-qun;LI Ju-yi;SHI Hui;NIU Yun-fei;FANG Zhao-hui(Department of Endocrinology,The First Affiliated Hospital,Anhui University of Traditional Chinese Medicine,Hefei 230038,Anhui Province,China;School of Nursing,Anhui University of Traditional Chinese Medicine,Hefei 230031,Anhui Province,China)

机构地区：[1]安徽中医药大学第一附属医院内分泌科,安徽合肥230038 [2]安徽中医药大学护理学院,安徽合肥230031

出　　处：《中国临床药理学杂志》2022年第17期2009-2012,共4页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金青年基金资助项目(81603602);安徽省自然科学基金资助项目(1908085MH267)。

摘　　要：目的 探究β-链蛋白(β-catenin)基因沉默对糖尿病血管钙化的抑制作用。方法 体外培养大鼠下肢股动脉内皮细胞。调整稳转细胞状态,分为空白组、阴性对照组、实验组和模型组。空白组给予常规培养;实验组用β-catenin慢病毒转染48 h后,给予钙化培养基培养12 d;阴性对照组用空载慢病毒细胞转染48 h后,给予钙化培养基培养12 d;模型组细胞在钙化培养基培养12 d,诱导细胞钙化。用实时荧光定量聚合酶链反应检测α平滑肌肌动蛋白(α-SMA)、β-catenin、碱性磷酸酶(ALP)和骨桥蛋白(OPN)mRNA的表达水平,用激光共聚焦检测细胞β-catenin蛋白定位。结果 病毒感染48 h后,空白组、阴性对照组和实验组的β-catenin mRNA分别为1.00±0.08,1.03±0.07和0.22±0.01。培养12 d后,空白组、模型组和实验组的α-SMA mRNA分别为1.00±0.03,3.68±0.23和1.99±0.10,β-catenin mRNA分别为1.00±0.07,4.51±0.16和2.15±0.15,ALP mRNA分别为1.00±0.09,4.02±0.13和2.30±0.12,OPN mRNA分别为1.00±0.08,5.32±0.50,3.12±0.27。实验组的上述指标与模型组比较,差异均有统计学意义(均P<0.01)。实验组核内β-catenin表达明显减少。结论 糖尿病大鼠下肢内皮细胞钙化发生机制是通过高糖诱导β-catenin mRNA翻译β-catenin蛋白在细胞核中的表达,从而激活下游基因的转录,沉默β-catenin基因可抑制钙化进程。Objective To explore the inhibitory effect of beta catenin(β-catenin) gene silencing on diabetic vascular calcification.Methods Endothelial cells were isolated and cultured in vitro.After adjusted to the stable state,the cells were divided into blank control group,model group,negative control group and experimental group.The blank control group was treated with conventional culture.The experimental group was transfected with β-catenin lentivirus for 48 h and then cultured in calcified medium for 12 days.The negative control group was transfected with empty lentivirus for 48 h and then cultured in calcified medium for 12 days.The model group was cultured in calcification medium for 12 days to induce cell calcification.The mRNA expression levels of α smooth muscle actin(α-SMA),β-catenin,alkaline phosphatase(ALP) and (OPN) were detected by reverse transcription-polymerase chain reaction.Cell β-catenin protein localization was detected by laser confocal detection.Results After 48 h virus infection,the results of β-catenin m RNA in blank,negative control and experimental groups were 1.00 ± 0.08,1.03 ± 0.07 and 0.22 ± 0.01,respectively.After 12 days of culture,the results of α-SMA m RNA in blank,model and experimental groups were 1.00 ± 0.03,3.68 ± 0.23 and 1.99 ± 0.10,β-catenin m RNA were 1.00 ± 0.07,4.51 ± 0.16,2.15 ± 0.15,ALP m RNA were 1.00 ± 0.09,4.02 ± 0.13,2.30 ± 0.12,OPN m RNA were 1.00 ± 0.08,5.32 ± 0.50 and 3.12 ± 0.27,respectively.The above indexes of the experimental group were compared with those of model group,and the differences were statistically significant( all P < 0.01).The expression of β-catenin in the nucleus was significantly decreased in the experimental group.Conclusion The mechanism of calcification in lower limb endothelial cells of diabetic rats is that high glucose inducesβ-catenin m RNA translation and β-catenin protein expression in the nucleus,thereby activating the transcription of downstream genes.Silencing β-catenin gene can inhibit the calcification proces

关 键 词：Β-链蛋白 糖尿病 内皮细胞钙化 基因沉默 

分 类 号：R97[医药卫生—药品]