鸢尾素对兔骨髓间充质干细胞成脂分化的影响

Effects of irisin on adipogenesis of bone-marrow mesenchymal stem cells from rabbits

目的探讨鸢尾素对兔骨髓间充质干细胞(bone-marrow mesenchymal stem cells,BMSCs)成脂分化的影响及其信号机制。方法分离、培养兔BMSCs,检测鸢尾素对BMSCs增殖的影响。BMSCs成脂诱导后,实时荧光定量PCR(polymerase chain reaction,PCR)检测成脂相关基因PPARγ和C/EBPα的转录表达,油红O染色检测细胞内脂质形成情况,Western blot检测Wnt10a信号蛋白的表达。结果鸢尾素不影响BMSCs的增殖活性;显著下调PPARγ[成脂诱导第7天,(0.20±0.06)vs(1.00±0.10);成脂诱导第14天,(0.61±0.06)vs(1.00±0.15)]和C/EBPα[成脂诱导第7天,(0.40±0.06)vs(1.00±0.12);成脂诱导第14天,(0.70±0.09)vs(1.00±0.18)]的表达,明显抑制BMSCs成脂分化[(0.45±0.05)vs(0.95±0.10);P<0.05];同时,鸢尾素明显促进Wnt10a表达[(5.01±0.78)vs(1.00±0.25);P<0.05]。结论鸢尾素可抑制BMSCs脂向分化,其作用与激活Wnt信号通路有关。

Objective To investigate the effects of irisin on adipocyte differentiation of BMSCs from rabbits and related mechanisms.Methods BMSCs of rabbits were extracted and proliferation capacity was detected. After BMSCs were cultured in adipogenesis medium supplemented with or without irisin, qPCR was used to evaluate the expression levels of PPARγ and C/EBPα, while oil red O staining was used to investigate fat accumulation. The signaling protein level of Wnt10 a was detected by Western blot.Results Proliferation capacity of BMSCs was hardly affected by irisin. Irisin could significantly downregulate mRNA expressions of PPARγ [(0.20±0.06) vs(1.00±0.10) 7 days after adipogenesis induction and(0.61±0.06) vs(1.00±0.15) 14 days after adipogenesis induction]and C/EBPα [(0.40±0.06) vs(1.00±0.12) 7 days after adipogenesis induction and(0.70±0.09) vs(1.00±0.18) 14 days after adipogenesis induction], while attenuating fat accumulation [(0.45±0.05) vs(0.95±0.10),P<0.05]. Moreover, Wnt10 a protein levels were increased by irisin intervention[(5.01±0.78) vs(1.00±0.25),P<0.05].Conclusions Irisin can inhibit adipocyte differentiation of BMSCs from rabbits, and this effect is related to activation of Wnt signaling pathway.