3. Shandong University of Traditional Chinese Medicine,Jinan 250355,China;Shandong Province Collaborative Innovation Center of Traditional Chinese Medicine Classics,Jinan 250355,China
Objective To observe the effect of serum containing Jinguishenqi pill on the osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs)in rats and to explore the mechanism of its occurrence.Methods The serum containing Jinguishenqi pill was prepared.CCK-8 method was used to screen out the effect of the optimal concentration of the serum containing Jinguishenqi pill on the osteogenic differentiation of BMSCs.BMSCs were divided into blank group(control group),osteogenic induction group(OIS),blank serum group(ROIS group),and drug-containing serum group(JKSQ-OIS).The osteogenic induction was carried out for BMSCs in each group respectively.After 14 or 21 consecutive days of intervention,alkaline phosphatase staining and Alizarin red staining were performed to observe the alkaline phosphatase activity and osteogenic mineralization level in each group under microscopic observation,respectively.After 14 consecutive days of intervention,real-time fluorescence quantitative PCR(q-PCR)and protein immunoblotting(Western blotting)were used to detect the mRNA and protein relative expressions of BMP-2,Smad1,ALP,Runx2,and OSX.Results Compared to the control group,the OIS and ROIS groups showed punctuate mineralized nodules,slightly darkened ALP staining,increased expressions of ALP and Runx2 mRNA and protein(P＜0.05,P＜0.01),and increased but not significant expression of OSX(P＞0.05).Compared to that in the OIS group,the mineralized nodules in the JKSQ-OIS group were in patches.ALP staining was obviously deepened,the expressions of BMP-2,Smad1,ALP,and Runx2 mRNA and protein increased(P＜0.05,P＜0.01),and the expressions of OSX mRNA and protein had a rising trend(P＞0.05).Conclusion The drug-containing serum of Jinguishenqi pill may promote the differentiation of BMSCs towards osteogenesis and relieve OP by regulating the BMP-2/Smad signaling pathway and up-regulating the expressions of ALP,Runx2,and OSX.
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