驻春胶囊通过调控miR-935促进成骨细胞分化的机制OACSTPCD

Objective:To investigate the effect of Zhuchun Capsule on osteoblast and the mechanism of Zhuchun capsule promoting osteo-blast differentiationby regulating miR-935.Methods:Mouse embryonic osteoblast progenitor cells MC3T3-E1 were culturedinvitro,and the cytotoxicity and concentration for 50%of maximal effect(EC50)of Zhuchun capsule were detected by CCK-8 method.MC3T3-E1 cells were divided into non-induction group,induction group,induction+Zuchun capsule group,and induction+Zhuchun capsule group.After 14 and 21 days of induction,bone formation induction of Mc3t3-e1 cells was observed by alizarin red staining.miR-935 was transfected into osteoblasts and the cells were divided into miR-935 inhibitor and inhibitor NC groups.miR-935,sig-nal transducer and activator of transcription 1 were detected by qRT-PCR and Western blot.STAT1),Runt-related transcription fac-tor 2(RUNX2),alkaline phosphatase(ALP),osteocalcin(OCN)gene and protein expression levels.Results:The EC50 value of Zu-chun capsule against MC3T3-E1 cells was4 684 mg·L-1.Alizarin red staining showed that different degrees of red nodules and cal-cium deposition were observed in the induced group,the induced+Zuchun capsule group and the non-induced+Zuchun capsule group compared with the non-induced group.qRT-PCR results showed that after 14 and 21 days of induction,compared with the non-induction group,the levels of ALP mRNA,RUNX2 mRNA,OCN mRNA and miR-935 mRNA in the induction group were increased,and the levels of STAT1 mRNA were decreased(P<0.05).The STAT1 mRNA level of cells in the non-induction+Zuchun capsule group was decreased(P<0.05).Compared with induction group,RUNX2 mRNA and OCN mRNA levels in induction+Zuchun cap-sule group were increased,while STAT1 mRNA levels were decreased(P<0.05).WB results showed that after14 and21 days of in-duction,compared with the non-induction group,the levels of ALP,RUNX2 and OCN in the induction group were increased,and the level of STAT1 was decreased(P<0.01);the levels of ALP,RUNX2 and OCN in the non-induction+Zuchun capsule group were in-creased,and the level of STAT1 was decreased(P<0.05).Compared with induction group,ALP,RUNX2 and OCN levels in the in-duction+Zhuchun capsule group were increased,while STAT1 level was checreased(P<0.05).qRT-PCR results showed that com-pared with normal control group,the mRNA level of miR-935 in Zhuchun capsule group was increased(P<0.01).Compared with in-hibitor NC group,OCN mRNA,ALP mRNA and miR-935 mRNA levels were decreased and STAT1 mRNA levels were increased in miR-935 inhibitor group(P<0.05).WB results showed that compared with inhibitor NC group,OCN and ALP levels of miR-935 inhibitor group were decreased,STAT1 levels were increased(P<0.05).Conclusion:Zhichun capsule can promote osteoblast differen-tiation by regulating miR-935,and its mechanism may be related to the expression of STAT1,RUNX2,ALP and OCN.