尿毒清胶囊人含药血清抑制尿毒症毒素血清刺激的肾小管上皮细胞-2增殖与转分化作用观察

Inhibitory Effect of Human Serum Containing Niaoduqing Capsule on Proliferation and Transdifferentiation of Human Renal Tubular Epithelial Cells

【目的】探讨尿毒清胶囊含药血清对人近端肾小管上皮细胞(HK-2)增殖与转分化的影响。【方法】将体外培养的HK-2细胞分为6组:正常对照组,体积分数5%、10%尿毒症毒素血清组(简称尿毒症血清组),体积分数5%、10%尿毒清胶囊含药血清组(简称含药血清组),蒙诺组(浓度为10-6mmol/L)。采用四氮唑蓝(MTT)法检测细胞增殖,流式细胞术检测细胞周期和α-平滑肌肌动蛋白(α-SMA)阳性细胞百分率。【结果】5%、10%尿毒症血清组光吸收值(D)显著高于正常对照组(均P<0.01)。5%含药血清组D值显著低于10%尿毒症血清组(P<0.01),10%含药血清和蒙诺组D值显著低于5%、10%尿毒症血清组(均P<0.01)。5%、10%尿毒症血清组G0/G1期细胞百分数(pG0/G1)显著降低(均P<0.01),细胞增殖指数(Ip)显著升高(均P<0.01)。5%、10%含药血清组及蒙诺组可显著升高pG0/G1,降低IP(均P<0.01)。5%、10%尿毒症血清组HK-2细胞α-SMA阳性细胞百分率(pα-SMA阳性)显著高于正常对照组(均P<0.01),5%含药血清组pα-SMA阳性显著低于10%尿毒症血清组(P<0.01),10%含药血清组和蒙诺组pα-SMA阳性显著低于5%、10%尿毒症血清组(均P<0.01)。【结论】尿毒清胶囊抗肾纤维化作用机制可能与其抑制参与肾纤维化的肾小管上皮细胞的增殖和转分化,进而抑制肾组织纤维化有关。

Objective To observe the effect of human serum containing Niaoduqing Capsule （NC） on proliferation and transdifferentiation of human renal tubular epithelial cells （ HK-2 ）. Methods The in-vitro cultured HK-2 were divided into 6 groups according to the culture media： normal control group, uremic serum groups （at the concentration of 5% and 10% ）, NC-containing serum groups （ at the concentration of 5% and 10% ）, and Monopril group （at the concentration of 10^-6mmol/L）. Cell proliferation was measured by methylene blue assay. Cell life cycle and the percentage of cells with a-smooth muscle actin （u-SMA） expression positive in every group were evaluated by flow cytometry. Results The optical absorption value in uremic serum groups was higher than that in the normal control group （P 〈0. 01 ） , lower in 5% NC-containing serum group than that in 10% uremic serum group （P 〈0. 01 ）, and lower in 10% NC-containing serum group and Monopril group than that in uremic serum groups （P 〈0. 01 ）. The percentage of cells in G0/G1 phase was remarkably lower and cell proliferation index was higher in uremic serum groups than those in the normal control group （ P 〈 0.01 ） , and serum containing NC and Monopril counteracted the above changes （ P 〈 0.05 or P 〈 0. 01 ）. The percentage of HK-2 ceils with α-SMA expression positive was increased significantly in uremic serum groups （P 〈 0. 01 compared with the normal control group） , was lower in 5% NC-containing serum group that that in 10% uremic serum group （P 〈0.01 ） , and lower in 10% NC-containing serum group and Monopril group than that in uremic serum groups （P 〈0. 01 ）. Conclusion The anti-renal-fibrosis mechanism of NC is probably related with the inhibition of proliferation and transdifferentiation of renal tubular epithelial cells.