银杏叶提取物对大鼠骨髓间充质干细胞成脂分化能力的影响

目的:探讨银杏叶提取物(GBE)对大鼠的骨髓间充质干细胞(BMMSCs)成脂分化能力的影响,阐明GBE在细胞水平上对骨质疏松治疗及预防的意义。方法:将50、100、150、200和400 mg·L^-1GBE与成脂诱导液加入到已纯化的第3代BMMSCs中(50、100、150、200和400 mg·L^-1GBE组),不加药的BMMSCs为对照组,共培养7 d,观察BMMSCs的形态学和表面抗原CD44、CD105和CD34的表达;采用油红O染色观察BMMSCs中脂滴形成情况,并对BMMSCs的成脂能力进行定量分析;采用RT-PCR法检测BMMSCs中脂肪组织脂肪酸结合蛋白(AP2)和过氧化物酶体增殖物激活受体(PPAR)的亚型(PPAR-γ)mRNA的表达水平。结果:形态学检测,体外培养的原代BMMSCs 3 d后可见细胞呈梭形,7 d后细胞呈漩涡状生长,同向排列;表面抗原标记物染色,CD44和CD105呈阳性表达,而CD34呈阴性表达,证明其为BMMSCs;油红O染色,约7 d后显微镜下见大量脂滴形成,并随着GBE浓度的增大脂滴数量减少;与对照组比较,其他各组BMMSCs的A₅₁₀值降低(P<0.05);RT-PCR检测,与对照组比较,其他各组细胞中AP2和PPAR-γ mRNA表达水平降低(P<0.05)。结论:GBE在体外可抑制BMMSCs的成脂分化能力,且抑制能力随着GBE浓度的增高而逐渐增强。

Effect of Ginkgo biloba extract on adipogenic differentiation ability of bone marrow mesenchymal stem cells in rats

Objective To study the effect of Ginkgo biloba extract (GBE) on the adipogenic differentiation ability of the bone marrow mesenchymal stem cells(BMMSCs),and to clarify its significance in treatment and prevention of osteoporosis at the cell level.Methods The third generation of BMMSCs were cultured with adipogenic induction solution(control group),and 50,100,150,200, and 400 mg·L^-1 GBE(50,100,150,200, and 400 mg·L^-1 GBE groups) for 7 d.The morphology of BMMSCs and the expressions of surface antisgens CD44,CD105, and CD34 were observed;the formation of lipid droplet in the BMMSCs was observed by Oil Red O staining;the quantitative analysis of the adipogenic ability of the BMMSCs was performed;RT-PCR method was used to test the expression levels of fatty acid-binding protein(AP2) and peroxisome proliferator activated receptor-γ(PPAR-γ) mRNA.Results The morphological detection results showed that the BMMSCs were spindle shaped after cultured in vitro for 3 d,and grew in whorls and arranged in the same direction after cultured for 7 d.The surface antigens CD44 and CD105 showed positive expression,and CD34 showed negative expression,which proved the BMMSCs were extracted correctly;the Oil red O staining results showed a large of lipid droplets which was seen under microscope,and the number of lipid droplets was reduced with the increasing of the concentrations of GBE; compared with control group,the A₅₁₀ values of the BMMSCs in other groups were decreased(P<0.05).The RT-PCR results showed the expression levels of AP2 and PPAR-γ mRNA were decreasd in other groups compared with control group (P<0.05).Conclusion GBE can inhibit the adipogenic differentiation ability of BMMSCs in vitro,and the inhibitory ability is enhanced with the increasing of GBE concentration.