P38MAPK抑制剂SB203580对高糖诱导HK-2细胞转分化的影响  

The effects of P38MAPK inhibitor SB203580 on TEMT of HK-2 cells

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

机构地区：[1]石河子大学第一附属医院肾病科,832008

出　　处：《天津医药》2016年第4期426-429,I0001,共5页Tianjin Medical Journal

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：目的探讨不同浓度P38丝裂原活化蛋白激酶(P38MAPK)抑制剂SB203580在高糖诱导肾小管上皮细胞-肌成纤维细胞转分化(TEMT)过程中的机制及其较佳作用浓度。方法体外培养人近端肾小管上皮细胞(HK-2)并分为对照组(5.5 mmol/L GS)、DMSO组(5.5 mmol/L GS+30μmol/L SB203580等体积的DMSO)、高糖组(30mmol/L GS),以及30 mmol/L GS+5、10、20、30μmol/LSB203580处理的S5、S10、S20及S30组,干预48 h。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);选取对照组、高糖组、S30组,Western blot法检测P38MAPK、P-P38MAPK及α-平滑肌肌动蛋白(SMA)的表达、免疫荧光法检测α-SMA的表达。结果 (1)与对照组相比,DMSO对HK-2细胞增殖无显著抑制作用(P>0.05);高糖组、S5组HK-2细胞增殖增多(P<0.05);S20、S30组HK-2细胞增殖减少(P<0.05)。与高糖组相比,S5、S10、S20、S30组细胞增殖均受到抑制(P<0.05)。(2)与对照组相比,高糖组、S30组P-P38MAPK表达量增高(P<0.05)。与高糖组相比,S30组P-P38MAPK的表达量降低(P<0.05)。3组P38MAPK表达量无显著差异(P>0.05)。(3)与对照组相比,高糖组、S30组α-SMA表达量增高(P<0.05)。与高糖组相比,S30组α-SMA表达量降低(P<0.05)。结论 30 mmol/L GS可以诱导HK-2细胞TEMT;30μmol/L SB203580是抑制HK-2细胞TEMT的较佳抑制浓度,SB203580可能通过下调P-P38MAPK表达,从而抑制HK-2细胞增殖及胞浆中α-SMA的表达,延缓TEMT进程。Objective To observe the effects of different concentrations of SB203580, the inhibitor of P38 MAPK, inprocess of high glucose（GS）-induced renal tubular epithelial-myofibroblast transdifferentiation（TEMT）. Methods Thecultured human renal tubular epithelial cells（HK-2） were divided into control group（5.5 mmol/L GS）, GS（30 mmol/L GS）group and different concentrations of SB203580（30 mmol/L GS ＋5, 10, 20 and 30 μmol/L SB203580） groups. The treat-ments were for 48 hours. MTT assay was used to observe cell proliferation. The median inhibiting concentration（IC50） was cal-culated. Western blot assay was used to detect the expressions of P38 MAPK, P-P38 MAPK and α-smooth muscle actin（α-SMA） in control group, high-glucose group and S30 group. The expression of α-SMA was also detected by the method of im-munofluorescence. Results 1.Compared with control group, there was no significant inhitory effect on proliferation rate inDMSO group（P 0.05）. There were increased HK-2 cells in high glucose group and S5group（P 0.05）. Proliferation rateswere significantly decreased in S20 and S30 groups（P 0.05）. Compared with high glucose group, the proliferation rates ofHK-2 cells were inhibited in S5, S10, S20 and S30 groups（P 0.05）. 2. The expression of P-P38 MAPK was significantlyhigher in high glucose group and S30 group than that of control group（P 0.05）. Compared with high glucose group, the ex-pression of P-P38 MAPK was significantly decreased in S30 group（P 0.05）, whereas no significant difference in the expres-sion of P38 MAPK between the two groups（P 0.05）. 3. Compared with control group, the expression of α-SMA was signifi-cantly increased in high glucose group and S30 group（P 0.05）. Compared with high glucose group, the expression of α-SMA was significantly decreased in S30 group（P 0.05）. Conclusion The 30 mmol/L GS can lead to TEMT in HK-2cells. The more suitable inhibitory concentration of SB203580 in the process of TEMT is 30μmol/L. SB203580 can slow

关 键 词：糖尿病肾病 肾小管 上皮细胞 上皮间质转分化 P38丝裂原活化蛋白激酶 SB203580 

分 类 号：R692[医药卫生—泌尿科学]