P38MAPK 抑制剂 SB203580对高糖诱导HK-2细胞转分化的影响

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 + 30 μmol/L SB203580 等体积的 DMSO）、 高糖组 （30mmol/L GS）， 以及 30 mmol/L GS +5、 10、 20、 30 μmol/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。

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

Abstract: Objective To observe the effects of different concentrations of SB203580, the inhibitor of P38MAPK, in process of high glucose (GS)-induced renal tubular epithelial-myofibroblast transdifferentiation (TEMT). Methods The cultured 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 P38MAPK, P-P38MAPK 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 in DMSO group (P > 0.05). There were increased HK-2 cells in high glucose group and S5group (P < 0.05). Proliferation rates were significantly decreased in S20 and S30 groups (P < 0.05). Compared with high glucose group, the proliferation rates of HK-2 cells were inhibited in S5, S10, S20 and S30 groups (P < 0.05). 2. The expression of P-P38MAPK was significantly higher 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-P38MAPK was significantly decreased in S30 group (P < 0.05), whereas no significant difference in the expres⁃ sion of P38MAPK 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-2 cells. The more suitable inhibitory concentration of SB203580 in the process of TEMT is 30μmol/L. SB203580 can slow down the process of TEMT by inhibiting P38MAPK activation and inhibiting proliferation and the expression of α-SAM s of HK-2 cells.