中成药2017，Vol.39Issue(12)：2462-2468,7.DOI:10.3969/j.issn.1001-1528.2017.12.004
补肾活血汤石油醚提取物对BMSCs迁移过程中Wnt5a/PKC通路的影响
Effect of the petroleum ether extract from Bushen Huoxue Decoction on promoting BMSCs migration via Wnt5a/PKC signaling pathway
摘要
Abstract
AIM To investigate the effect and mechanism of petroleum ether extract from Bushen Huoxue Decoction (BSHXD,Rehmanniae Radix Praeparata,Cuscutae Semen,Psoraleae Fructus,etc.) on rat BMSCs migration.METHODS Bone marrow mesenchymal stem cells isolated from whole bone marrow of rats were amplified by adherent culture in vitro.Flow cytometry was employed to detect the cell surface antigens of the third generation cells.Migration examination of cells treated with petroleum ether extract from BSHXD at concentrations of 25,50,100 and 200 μg/mL was accomplished by cell scratch test and Transwell assay.Expressions of WntSa and PKC protein were traced by Western blot,and so were the mRNA expressions of Wnt5a and PKC by RT-PCR.RESULTS Positive expressions of CD29,CD44 and CD90 in the passage 3 of BMSCs but negative expression of CD45 were observed.The cells given different concentrations of petroleum ether extract from BSHXD groups displayed a significantly higher wound healing speed than the control group after 6 h and 12 h cell scratch (P ＜ 0.05).100 μg/mL BSHXD was shown to be the most active concentration for promoting cell migration by Transwell migration experiments,with a statistically significant difference (P ＜ O.O1).Compared with the control group,the levels of Wnt5a mRNA in the BSHXD groups were significantly higher (P ＜ 0.05),and the PKC mRNA level in 100 μg/mL petroleum ether extract from BSHXD group was significantly increased as well (P ＜0.01)by RT-PCR.Although BSHXD groups brought forth significant improvement in both the expressions of Wnt5a and PKC protein (P ＜ 0.01),the 100 μg/mL concentration conferred the highest (P ＜ 0.01).CONCLUSION Petroleum ether extract from BSHXD can promote BMSCs migration in vitro,and the mechanism may lie in its association with WntSa signaling pathway.关键词
补肾活血汤/信号通路/体外迁移/骨髓间充质干细胞/Wnt5a/PKCKey words
Bushen Huoxue Decoction/signaling pathway/migration in vitro/bone marrow mesenchymal stem cells/Wnt5a/PKC分类
医药卫生引用本文复制引用
罗辉,吴志方,沈玮,胡流超,王斌,余丽娟,罗毅文..补肾活血汤石油醚提取物对BMSCs迁移过程中Wnt5a/PKC通路的影响[J].中成药,2017,39(12):2462-2468,7.基金项目
国家自然科学基金(81473699,81273783) （81473699,81273783）
广东省自然科学基金(2016A030310289) （2016A030310289）
