整合素连接激酶抑制剂QLT0267对TEMT过程中P38MAPK表达的影响  

The effect of QLT0267 which is inhibitor of ILK on P38MAPK in process of HK-2 cells TEMT

作　　者：林智峰[1] 贾林[2] 赵丹[1] 马莉[2] 唐玉玲[2] 杨锐[2] 杨晓萍[1] 

机构地区：[1]石河子大学医学院第一附属医院肾病科,新疆石河子832008 [2]石河子大学医学院,新疆石河子832008

出　　处：《重庆医学》2016年第23期3206-3209,3212,共5页Chongqing medicine

基　　金：石河子大学科学技术研究发展计划基金资助项目(2013ZRKXYQ-YD16)

摘　　要：目的探讨在高糖诱导肾小管上皮细胞-肌成纤维细胞转分化(TEMT)过程中,整合素连接激酶(ILK)抑制剂QLT0267对P38丝裂原活化蛋白激酶(P38MAPK)表达的影响。方法体外培养人近端肾小管上皮细胞(HK-2),分10组,每组给予不同浓度葡萄糖及QLT0267,干预48h。四甲基偶氮唑蓝(MTT)法检测细胞增殖情况,计算半数抑制浓度(IC50);随后选取对照组、高糖组、DMSO组、QLT0267(IC50)组(使用QLT0267后,HK-2细胞数量减少一半所需的QLT0267药物浓度),免疫荧光法检测各组ILK、P-P38MAPK的表达;Western blot检测各组ILK、蛋白激酶B(AKT)、磷酸化-蛋白激酶B(P-AKT)、PP38MAPK、P38MAPK、α-平滑肌肌动蛋白(α-SMA)的表达,观察各组表达差异性,探讨在TEMT过程中QLT0267对P38MAPK的影响。结果 30mmol/L的葡萄糖干预48h HK-2细胞增殖显著,同时上调细胞中ILK、P-AKT、P-P38MAPK、α-SMA的表达;QLT0267在抑制HK-2细胞增殖的同时,可以通过下调ILK下游P-AKT的表达,阻止P38MAPK活化,进而减少HK-2细胞中α-SMA的表达,延缓TEMT进程。结论 QLT0267可能通过部分阻止P38MAPK的活化,从而延缓HK-2细胞TEMT进程。Objective To observe the effect of different concentrations of QLT0267 which is the inhibitor of integrin-linked kinase（ILK） on P38 mitogen activated protein kinase （P38MAPK）,in process of high glucose-induced tubular epithelial-myofibro blast transdifferentiation （TEMT）. Methods The cultured human renal tubular epithelial（HK-2） cell were divided into 10 groups by the different concentrations of GS and QLT0267 ,cultured 48 h. The inhibition ratio in each group was calculated by the method of MTT,and found concentration which inhibit rate was 50% （IC50）. Then we selected control group, high-glucose group, DMSO group and QLT0267 （IC50）group（The concentration of QLT0267, which led to the HK-2 cell reduced by half）. The protein of ILK, AKT,P-AKT,P38MAPK,P-P38MAPK and s-smooth muscle actin （α-SMA） were detected by the method of immunofluorescence and Western blot. We observed the differences in each group and explored the relationship between ILK and P38MAPK in the process of TEMT. Results HK-2 cell proliferated significantly after 30 mmol/L glucose intervention for 48 h,and at the same time it upregulated the expression of ILK,P-AKT, P-P38MAPK,α-SMA. 50% HK-2 cell inhibited at the concentration of 28 μmol/L QLT0267. It decreased both the expression of P-AKT which down-regulated the expression of ILK, and the expression of P- P38MAPK, thereby reducing the expression of α-SMA and delaying process of TEMT at the concentrations of 28 μmol/L QLT0267 in HK-2 cell. Conclusion 28 μmol/L QLT0267 might inhibit HK-2 cell TEMT by decreasing the expression of P38MAPK.

关 键 词：糖尿病肾病 近端小管上皮细胞 转分化 整合素连接激酶 P38丝裂原活化蛋白激酶 QLT0267 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]