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.