丹酚酸B对高糖干预下的人肾小管上皮细胞自噬相关蛋白的影响

Effect of salvianolic acid B on high glucose under the intervention of human renal tubular epithelial cell autophagy related protein

目的探究丹酚酸B对高糖干预下的人肾小管上皮细胞自噬相关蛋白的影响。方法根据人近端肾小管上皮细胞HK-2细胞培养方式的不同,分为对照组、葡萄糖组和实验组,对照组加入5.5mmol/L的葡萄糖;葡萄糖组加入葡萄糖的浓度分为10、20和30mmol/L;实验组在培养基中分别加入丹酚酸B和葡萄糖(30mmol/L),丹酚酸B的浓度分为0.1、1、10和100μmol/L,培养24h。使用蛋白提取试剂盒分别提取3组培养细胞中的总蛋白,采用Bradford法测定不同培养组细胞自噬相关蛋白的含量,进行分析比较。结果随着葡萄糖刺激浓度的增高,p62蛋白表达增加。24h后,高糖可诱导HK-2细胞自噬相关蛋白p62表达增加,LC3表达减少,且呈剂量依赖性。与对照组相比,随着丹酚酸B干预剂量的增高,24h后的细胞LC3的表达增加,不同剂量丹酚酸B干预可减少高糖诱导下的p62蛋白的表达增加,同时LC3的表达得到明显恢复,且呈剂量依赖性(P<0.05)。Logistic回归分析的结果显示,LC3蛋白含量、p62蛋白含量是影响细胞自噬的独立影响因素。结论在高糖的干预下,不同剂量丹酚酸B干预可减少高糖诱导下的p62蛋白的表达增加,同时LC3的表达得到明显恢复,且呈剂量依赖性。

Objective To explore the effect of salvianolic acid B on high glucose under the intervention of human renal tubular epithelial cell autophagy related protein.Methods According to the cell culture in different ways,divided into control group,D-glucose stimulation group and salvianolic acid B＋D-glucose stimulation group.The control group was to join the 5.5mmol/L concentration of glucose.Glucose stimulated D-group added glucose was divided into 10mmol/L,20mmol/L,30mmol/L.Salvianolic acid B＋D-glucose stimulation group in the cultivation of salvianolic acid B and D-were added to the medium（glucose concentration is 30mmol/L）,the concentration of salvianolic acid B was divided into 0.1mol/L,1mol/L,10mol/L,100mol/L.The use of protein extraction kit to extract cell total protein content,for different groups of cells autophagy related protein culture carries on the analysis comparison.Results With the increase of the concentration of glucose stimulation was increased expression of p62.High glucose could induce autophagy in HK-2cell associated protein p62 expression increased,LC3 expression decreased,and showed dose dependent after 24 hours.Compared with the control group,with the increase of salvianolic acid B intervention dose,the expression of cell LC3-II increased gradually,different doses of salvianolic acid B intervention can reduce the expression of high glucose induced p62 protein increased,while the expression of LC3 was obviously restored,and showed dose dependent.There were statistical significantly differences（P〈0.05）.The results of Logistic regression analysis showed,LC3 content of II protein,p62 protein content was the independent factors on autophagy.ConclusionIn high glucose under the intervention of different doses of salvianolic acid B intervention can reduce the expression of high glucose induced p62 protein increased,while the expression of LC3 was obviously restored and showed dose dependent.