Study On The Mechanism Of Maimendong Decoction To Promote The Migration Of Rat BMSCs In Vitro Via SDF-1/CXCR4 Axis

Objective In vitro observation was conducted to study whether Maimendong Decoction affects the migration of rat bone marrow mesenchymal stem cells(r BMSCs)through the SDF-1/CXCR4 axis.Method The whole bone marrow adherence method was used to isolate and culture r BMSCs.Flow cytometry was used to detect the expression of CD34,CD44,and CD105 antigens on the surface of r BMSCs and to identify the purity of r BMSCs;CCK8 method was used to detect the effect of Maimendong Decoction on the proliferation of r BMSCs,and to screen the optimal intervention concentration And time;scratch experiment and Transwell experiment to detect the effect of Maimendong Decoction on the migration of r BMSCs;real-time fluorescent quantitative PCR and Western Blot to detect the expression of r BMSCs SDF-1 and CXCR4 genes and proteins.Results The whole bone marrow adherent method can obtain r BMSCs with high purity,uniform morphology and good condition.Flow cytometric detection of r BMSCs surface antigen CD34 positive rate was 0.6%,CD44positive rate was 99.4%,and CD105 positive rate was 99.7%.It conforms to the identification of r BMSCs surface antigen,and the purity of r BMSCs cells is high,which meets the requirements of subsequent experiments;CCK8method found that different concentrations of Maimendong decoction extracts can promote the proliferation of r BMSCs,however,the 24 h cell viability and proliferation rate of Maimendong decoction with a concentration of3.125mg/L is the best(P <0.05);the scratch test and Transwell experiment found that compared with the blank group,Maimendong decoction has the effect of promoting cell migration(P <0.05),while CXCR4 receptor blockade the retarder ADM3100 inhibited the migration of r BMSCs(P <0.05);compared with the blank group,the r BMSCs pretreated with Maimendong decoction had high expression of SDF-1 and CXCR4 m RNA(P <0.05),while ADM3100 inhibited the migration of r BMSCs.r BMSCs SDF-1,CXCR4 m RNA expression(P <0.05);Western Blot detection showed that compared with the blank group,the pretreatment of Maimendong decoction could up-regulate the expression of r BMSCs CXCR4 protein(P <0.05),while ADM3100 inhibited the expression of r BMSCs CXCR4 protein(P <0.05).Conclusion 1.The whole bone marrow adherence method is simple and efficient to isolate and extract r BMSCs,with high cell purity,uniform morphology and good condition;2.Maimendong decoction extract has the effect of improving the cell activity of r BMSCs,promoting the proliferation and migration of r BMSCs in vitro,and the concentration is 3.125 mg/lintervention 24 h cell activity and proliferation rate is the best;3.Maimendong Decoction can up-regulate the expression levels of r BMSCs SDF-1 and CXCR4,suggesting that Maimendong Decoction promotes the in vitro migration of r BMSCs related mechanisms may be related to the up-regulation of r BMSCs SDF by Maimendong Decoction-1 and CXCR4 expression is up-regulated,which activates the SDF-1/CXCR4 biological axis.