The Effect Of Psoralen On The Osteogenic Differentiation Of Rat Bone Marrow Mesenchymal Stem Cells And Its Mechanism

Purposes:Observing the effect of psoralen on the osteogenic differentiation of rat bone marrow mesenchymal stem cells cultured in vitro and the effect on the expression of TGF-β1and Smad4 proteins to elucidate the psoralen regulation of bone metabolism and prevention and treatment of osteoporosis Possible mechanism of action.Material and methods:Rat bone marrow mesenchymal stem cells were isolated and extracted using rat femur bone marrow adherent method.After subculturing three times,the cells were divided into blank control group,condition-induced group,psoralen group and psoralen-conditioned condition-induced group.Blank control group: 10% FBS in DMEM culture medium;conditional induction group: osteogenic inducer culture;psoralen group:culture containing 15umol/L psoralen for culture;psoralen combined condition induction Group: Incubation with an osteogenic inducer containing 15 umol/L psoralen.During this period,the growth status of the cells was observed at every moment,and the changes were made once every 3 days.After 12 days,various palliative tests were performed.Elisa method was used to detect the expression level of ALP in the supernatant of each group;RT-PCR was used to detect the expression of chemokine CXCL12;Western Blot was used to detect the expression of COL I,Cbf ɑ1,TGF-β1,Smad4 protein.Results:1.Elisa method showed that compared with the blank control group,the levels of ALP in the medium of condition-induced group,psoralen group and psoralen-condition-induced group were significantly higher(p<0.05),and conditional induction Compared with the group,the level of ALP in the psoralen combined condition-induced group was significantly higher(p<0.05).2.RT-PCR showed that the levels of CXCL12 mRNA in the condition-induced group,psoralen group,and psoralen-induced condition group were significantly higher than those in the blank control group(p<0.05).The ratio of CXCL12 mRNA in the psoralen combined condition-induced group was significantly higher(p<0.05).3.WesternBlot method: Compared with the blank control group,the levels of COL I,Cbf ɑ1,TGF-β ?1,Smad4 in condition-induced group,psoralen group,and psoralen-induced condition group were significantly higher(p< 0.05)Compared with the condition-induced group,the levels of COL I,Cbf ɑ 1,TGF-β 1,Smad4 were significantly higher in the psoralen combined condition-induced group(p<0.05).Conclusion:1.Psoralen can promote the expression of ALP,CXCL12,COL I and Cbf ɑ1 in rat bone marrow mesenchymal stem cells and induce their differentiation into osteoblasts.2.The mechanism of prevention and treatment of osteoporosis by psoralen may be through affecting the TGF-β 1/Smad4 signaling pathway,so that synergistic osteogenic inducer promotes the differentiation of rat bone marrow mesenchymal stem cells into osteoblasts.