当归芍药散含药血清通过调节巨噬细胞活化诱导的肾小管上皮细胞转分化发挥肾保护作用

Danggui Shaoyao San drug-containing serum plays a protective role by regulating the transdifferentiation of renal tubular epithelial cells induced by macrophage activation

目的观察当归芍药散含药血清对肾小管上皮细胞转分化的干预作用。方法制备当归芍药散含药血清(DSS)。LPS活化巨噬细胞J774,取巨噬细胞培养上清液作为肾小管上皮细胞NRK-52E培养基,刺激肾小管上皮细胞转分化。实验设正常组,模型组,当归芍药散含药血清组。细胞培养24h,48h后,MTT法检测NRK-52E增殖状况,倒置相差显微镜观察NRK-52E细胞形态,RT-PCR检测各组NRK-52E细胞α-平滑肌肌动蛋白(α-SMA)和细胞角蛋白19(CK19)mRNA表达,免疫细胞化学法观察NRK-52E细胞E-钙黏素(E-cadherin)表达变化。结果与正常组比较,模型组肾小管上皮细胞形态出现明显改变,细胞形态拉长呈梭形,细胞之间排列欠紧密;与模型组相比,当归芍药散含药血清组上述细胞形态变化程度减轻。与模型组比较,当归芍药散含药血清组OD值无统计学意义;24h,48hα-SMA mRNA水平降低,CK19mRNA水平升高(P<0.05);48h E-cadherin表达水平升高(P<0.01)。结论当归芍药散含药血清可能通过调节巨噬细胞活化诱导的肾小管上皮细胞转分化发挥肾保护作用。

Objective To observe the interventional effect of Danggui Shaoyao San(DSS)medicine serum on the transdifferentiation of renal tubular epithelial cells.Methods DSS medicine serum was prepared.Macrophages J774 was activated by LPS.To stimulate the transdifferentiation of renal tubular epithelial cells,the activated macrophage culture supernatant was extracted as NRK-52 E cell culture medium.NRK-52 E was divided into three groups:Control group,Model group and DSS medicine serum group.After the cells were cultured for 24 h and 48 h,the proliferation status of NRK-52 E was detected by MTT,NRK-52 E cells morphology was observed by inverted phase contrast microscope,α-SMA and CK19 mRNA expression were detected by RT-PCR,E-cadherin expression in NRK-52 E cells was detected by Immunocytochemistry.Results Compared with the normal group,the morphology of NRK-52 E in the model group changed significantly,the cell morphology was elongated and showed a spindle shape,and the cells were not arranged closely;Compared with the model group,the above-mentioned cell morphology changes in DSS medicine serum group were reduced.Compared with the model group,the OD value of DSS medicine serum group was not statistical difference;24 h and 48 hα-SMA mRNA level was decreased,CK19 mRNA level was increased(P<0.05);48 h E-cadherin expression was increased in Immunocytochemistry(P<0.01).Conclusion DSS medicated serum may play a protective role to kidney by regulating the transdifferentiation of renal tubular epithelial cells.