Influences Of Black Cohosh Extract On Osteogenetic Differentiation From Bone Marrow Mesenchymal Stem Cells Of Rats Through The ER/NO/cGMP Signal Pathway

ObjectiveIn this article, we tested the osteogenetic differentiation index and the molecule of ER/NO/c GMP signal pathway of rat bone marrow mesenchymal stem cells(BMSCs) that was under the intervention of black cohosh extract(Actaea racemose, AR) in vitro. Finally, we confirmed that AR is an effective drug for the treatment of Postmenopausal osteoporosis(Postmenopausal osteoporosis,PMOP) and speculated that the ER/NO/c GMP signal pathway may be associated with the treatment mechanism of AR. MethodWe got BMSCs from the SD rat bone marrow cavity and carried on culturing cells. The surface antigen and purity of the third generation of cells was identified by Fluid cytology technology. Finally, the third generation of BMSCs was induced by the osteogenetic differentiation medium.Added AR(10-6-10-11mol/L) and 17 beta estradiol(E2, 10-8mol/L) into the osteogenetic differentiation medium respectively, processed BMSCs, and detected ALP activity. At the same time, we determined the optimal concentration of AR.Added AR(10-9mol/L), E2(10-8mol/L), estrogen receptor blockers(10-6mol/L) and nitric oxide synthase inhibitors(6x10-3mol/L) into the osteogenetic differentiation medium respectively, processed r BMSCs and measured the levels of osteogenesis differentiation index and the molecule of the signal pathway. ResultThe third generation of BMSCs was presented a uniform long spindle shape and closely arranged a spiral shape. Via Fluid cytology technical appraisal, the expressions of third generation of cell surface antigens were positive to CD90(100%), CD29(99.9%) and negative to CD45(0%), CD11b(0.4%).After the 21 days’ induction, it presented mineralized nodules with Vonkossa staining, which suggested the success of osteogenetic differentiation.AR(10-6-10-11mol/L) increased ALP activity in a dose-dependent manner and was significantly higher than that of the control group(P < 0.05). At a concentration of 10-9mol/L, ALP activity was optimal, which had no significant differences compared with that of E2(P > 0.05).AR(10-9mol/L) and E2(10-8mol/L) can cause the increase of the contents of TNOS、ALP、Calcium deposits、NO、c GMP and the expressions of ALP m RNA、e NOS m RNA、i NOS m RNA, and both was significantly higher than that of the control group(P < 0.05). These effects both can be blocked by ICI182, 780 and L-NAME, and the roles of AR and E2 had no significant differences(P > 0.05). Conclusion 1. AR can promote osteogenesis differentiation of BMSCs. At a concentration of 10-9mol/L, ALP activity was optimal, which had no significant differences compared with that of E2. So we confirm that AR can treat PMOP effectively. 2. Compared with E2 and ER/NO/c GMP signal channel blockers intervention, we confirm the ER/NO/c GMP signal pathway is one of the molecular mechanisms during the treatment of PMOP with AR. 3. Look for a safe and effective drug for treatment of PMOP, and provide potential therapeutic targets for diversely treating PMOP.