黄芪甲苷孵育脂肪源性干细胞对顺铂诱导的肾小管上皮细胞损伤的影响

Effect of adipose- derived stem cells cultured with astragaloside IV on the cisplatin- inducedrenal tubular cells

目的观察黄芪甲苷（Ast）孵育后的脂肪源性干细胞（ADSC）对顺铂诱导的肾小管上皮细胞（HKC）损伤的影响。方法HKC被分为4组：（1）对照组；（2）顺铂组；（3）ADSC与损伤HKC共培养组（ADSC＋HKC组）；（4）Ast孵育ADSC后与损伤HKC共培养组（Ast＋ADSC＋HKC组）。分别用不同浓度的顺铂（2．5、5、10、20μmol／L）诱导HKC6、24h，建立肾小管细胞损伤模型。用Transwell小室与20mg／LAst孵育的ADSC间接共培养48h。流式细胞术（FCM）和原位末端转移酶标记法（TUNEL）检测HKC的细胞凋亡；以增殖细胞核抗原（PCNA）反映HKC增殖情况；ELISA法检测下室HKC培养液中白细胞介素6（IL－6）、T细胞表达和分泌因子（RANTES）、红细胞生成素（EPO）、类胰岛素一号增长因子（IGF－1）的表达；结晶紫染色观察上室膜下ADSC的迁移指数。结果FCM和TUNEL结果均显示，与顺铂组相比，ADSC＋HKC组和Ast＋ADSC＋HKC组细胞凋亡的比例和数量减少，差异有统计学意义（均P〈0．05）；PCNA染色结果显示ADSC＋HKC组和Ast＋ADSC＋HKC组的细胞增殖数量高于顺铂组，差异有统计学意义（P〈0．05）；与顺铂组相比，Ast＋ADSC＋HKC组IL－6、RANTES水平明显降低，EPO、IGF－1水平升高，差异均有统计学意义（均P〈0．05）；结晶紫染色显示Ast＋ADSC＋HKC组ADSC的迁移指数高于其他3组（均P〈0．05）。结论黄芪甲苷可刺激ADSC的旁分泌效应，增强ADSC向受损部位定向迁移，从而减少HKC凋亡。

Objective To investigate the role of human adipose-derived mesenchymal stem cells （ADSC） cultured with astragaloside IV（Ast） on renal tubular epithelial cells line （HKC） induced by eisplatin in vitro. Methods HKC was divided into four groups： （1） normal control group; （2） cisplatin group; （3） ADSC and HKC group; （4） ADSC cultured with Ast and HKC group. After coincubated with ADSC,the apoptosis index and proliferation of HKC were detected respectively. The levels of interleukin- 6 （IL- 6）, regulated upon activation normal T cell expressed and secreted （RANTES）, erythropoietin （EPO） and insulin like growth factor 1 （IGF- 1） were detected by ELISA. Transwell culture system were used to test the migration effect of ADSC. Results Compared with cisplatin group, the apoptotic rates of HKC were decreased and the cell number increased obviously （P 〈 0.05） in ADSC and HKC group and ADSC cultured with Ast and HKC group. Compared with cisplatin group, the expression of IL-6 and RANTES were lower （P 〈 0.05）, and the level of EPO andIGF- 1 increased significantly （P 〈 0.05）, in ADSC cultured with Ast and HKC group. Crystal violet staining showed that ADSC cultured with Ast and HKC group migration index was higher than the other three groups （P 〈 0.05）. Conclusion The intervention of Ast may stimulate paracrine effect and migration of ADSC, so as to reduce apoptosis of HKC.