| Objective: To establish culture procedures of bone marrow stromal cells from SD rats, observetheir growth and characteristics and investigate the effects of osthole on their proliferation andadipocyte of differentiation ability. To investigate the expression of leptin in induced system withosthole in molecular biology leval. The study may provide a better understanding of themechanisms involved in the pathogenesis of OP and another aim of this study is to illuminate thebiological property of osthole fortunei on OP.Methods:1. MSCs from SD rats were isolated and purified by using differential attachment method, thenidentified according to morphology, induced-differentiation potentiality and the cell surfaceantigens. So we can establish a stable culturing system in viro;2. MTT method was used to investigate the proliferation effects osthole on MSCs in SD rats,and to determine the optimal concentration of osthole;3. Oil Red O staining was used to study the effects of osthole on adipogenic differentiation ofMSCs, and to determine the optimal concentration of osthole;4. The third passage MSCs were divided into control group, induced group and osthole group,to illuminate the effects of osthole on the adipogenic transdifferentiation of MSCs. Theexpression level of ALP was detected with PNPP method, the content of TG was detectedwith GPO-PAP method;5. Semi-quantitative analysis was used to detect the expression of Leptin during adipogenicdifferentiation through RT-PCR method. Grouping was the same as above.Results:1. Primary cultured MSCs adhered to plastic surface within 24 hours and reached confluencewithin 10-14days. To be conditioned induced, MSCs were successfully differentiated intoosteoblast and adipocyte. And confirmed that there was expression of CD29 and CD44, andno expression of CD34 and CD45 on the surface of MSCs by flow cytometry (FCM).2. OD values of MTT shows osthole groups of 1×10-7, 1×10-6, 1×10-5mol/L were higer thanblank groups, significantly (P＜0.05), especially 1×10-6mol/L.3. The number of adipocytes shows osthole groups of 1×10-8, 1×10-7, 1×10-6mol/L were lowerthan induced groups, significantly (P＜0.05),especially 1×10-7mol/L. 4. Alkaline phosphatase (ALP) activity values in MSCs treated with adipogenetic inductionmedia and osthole 12 days of osthole groups were higher than that in blank groups andinduced groups. There were significant differences between osthole groups and the other 2groups (P＜0.001). But there were no significant differences between induced groups andblank groups (P＞0.05).5. The contents of triglyceride in the cells in induced groups were higher than that in blankgroups and osthole groups at 12 days. There were statistically significant differences betweeninduced groups and the other 2 groups (P＜0.001). And there were not statistically significantdifferences between Group blank and osthole (P＞0.05).6. The expression of the Leptin mRNA in the cells in osthole groups were higher than that inblank groups and induced groups (P＜0.05), There was no significant differences betweengroup blank and induced (P＞0.05).Conclusion:1. Osthole can promote the proliferation of primary mouse bone MSCs;2. Osthole inhibited differentiation of MSCs into adipocytes induced by adipogenetic agonist;3. Osthole increased the ALP activity values and decreased the contents of triglyceride;4. Osthole increased the expression of LeptinmRNA, inhibited the character of differentiationof MSCs into adipocytes. |
