摘要
目的:探讨体外培养中I型磷酸酶的新型抑制亚基(IPP5)对高糖高胰岛素诱导的心肌细胞的肥大是否具有抑制作用。方法:新生SD乳鼠心肌细胞在含有10%胎牛血清的杜尔伯科改良伊格尔培养基(DMEM)中培养72h后,分别将IPP5野生型和短片段活化突变型基因通过脂质体介导转染入心肌细胞,48h后,加入高糖高胰岛素作用48h。利用图像分析软件分析心肌细胞表面积,并检测心肌细胞总蛋白含量、肌酸激酶(CK)和乳酸脱氢酶(LH)漏出量及心肌损伤标志物心肌肌钙蛋白I(TnI)的变化。结果:高糖高胰岛素可增加心肌细胞表面积,IPP5突变活化型基因可部分抑制心肌细胞肥大;转染IPP5突变活化型基因组的心肌细胞总蛋白含量较转染空载体对照组高,而CK和LH漏出量及TnI的含量却比转染空载体对照组明显减少。此外,转染IPP5突变活化型基因组的心肌细胞形态轮廓清楚,细胞质颗粒少,而转染空载体对照组的心肌细胞轮廓不清,细胞质颗粒粗大且多。结论:IPP5突变活化型对高糖高胰岛素诱导的心肌细胞肥大、损伤具有一定的抑制保护作用。
Objective:To determine whether the inhibitor-2 of protein phosphatase I(IPP5) has an inhibitory effect on cardiomyocyte hypertrophy induced by high glucose and insulin in neonatal rats.Methods:Cardiomyocytes of neonatal SD rats were cultured in DMEM containing 10% calf bovine serum for 72 hours.Plasmids containing wild type or activated mutant of IPP5 gene were transfected into cardiac myocytes by lipofectamine respectively.After 48 hours,cardiomyocytes were treated with high glucose plus high insulin for another 48 hours.Then image analysis software was used to analyse the surface area of cardiomyocytes,and the concentration of total protein,creatine kinase(CK),lactate dehydrogenase(LH).The injured markers of cardiomyocytes TnI in the supematant of cell culture were determined.Results:The cellular surface area was significantly increased after treatment with high glucose and insulin compared with the control.The activated mutant of PPI2 gene partially inhibited the increased celular surface area induced by high glucose and insulin.After high glucose and insulin treatment,the total protein content of activated mutant IPP5 gene transfected cardiomyocytes was significantly increased than that of mock-transfected cardiomyocytes,while the concentration of CK,LH and TnI was significantly decreased.In addition,in the activated mutant gene-transfected cardiomyocytes,the figure of cells was clear and the granules in cytoplasm was few and small,but in the mock-transfected cardiomyocytes,the figure was not clear,and it presented much larger granules in cytoplasm.Conclusion:The activated mutant of IPP5 gene might have inhibitory effects on cardiomyocyte hypertrophy induced by high glucose and insulin,and protect cardiomyocytes from injury by high glucose and insulin.
出处
《广西医科大学学报》
CAS
2010年第5期672-675,共4页
Journal of Guangxi Medical University
基金
广西自然科学基金(No.0542077)
