Effects Of Shengmai-injection On Multiple Hematopoietic-Related Factors Manifested By BMSCs Of SD Rats

Objective: The Method of isolation and culture of the SD rats bone mesenchymalstem cells (BMSCs) in vitro were discussed, appraisaling their biological characteristics,the experimental foundation was offered for the further research and practical clinicalapplication. The effect of shengmai injections on the promoting proliferation and secretionhematopoietic growth factor for the SD rat BMSCs were discussed, thus the powerfulexperimental basis could be provided for the shengmai injection in clinical application ofblood disease.Methods: The SD rats were used for femur and tibial marrow, the mesenchymal stemcells(MSCs) were isolated and cultured by using the method of adhesive screening, thenthe cell morphological characteristics of MSCs were observed under the microscope, andthe biological characteristics of BMSCs were identified through the flow cytometrictechnology. The effection of shengmai injection on the proliferation of BMSCs wasdetected with MTT colorimetric method, the expression of hematopoietic growth factorsand effection of shengmai injection on the expression of hematopoietic growth factorswere detected with Real Time-PCR method.Results: After the BMSCs were inoculated in12~24hours, most cells can stick to thewall, along with the extending of time, visible the colony growth, presented thefibroblast-like, the surface markers of BMSCs was identified by flow cytometry: thepositive rate of CD29is96.57%, the positive rate of CD44is93.63%, the positive rate ofCD34is3.6%, the positive rate of CD45is12.3%. The expression of the GM-CSF, TPOand SCF were existence; After culture different concentrations of shengmai injection andBMSCs24h,48h,72h, the proliferation effect of containing5μL/ml concentration ofshengmai injection in group48h is the most obvious(P＜0.05), certifing the culturemedium contains5μL/ml concentration of shengmai injection is the most suitable for theBMSCs proliferation; The Real Time-PCR analysis shows the BMSCs can secrete GM-CSF, SCF and TPO, and the expression of the mRNA of GM-CSF and SCF issignificantly higher than the control group(P＜0.05), the expression of the mRNA of TPOdeclined obviously compared with the control group, and with the ninth most obvious, thedifferences had the significant(P＜0.05).Conclusions:1. The BMSCs could be separated successfully from the SD rat bone marrow by usingthe method of adhesive screening, and could detect many hematopoietic growth factors.2. The shengmai injection can promote BMSCs born in vitro proliferation, and canpromote the BMSCs secrete the GM-CSF and SCF, this may reveal the BMSCs couldpromote hematopoietic role mechanism.3. The shengmai injection can restrain the BMSCs secrete the TPO, this may revealthe shengmai injection has the mechanism of promoting blood circulation and removingblood stasis and anticoagulant effect.