【作者】 林春淑； 孟晓军； 肖菲娜； 容婵； 舒晓春；

【Author】 LIN Chun-Shu;MENG Xiao-Jun;XIAO Fei-Na;Department of Health Management Center,The Fifth Affiliated Hospital of Sun Yat-sen University;

【机构】 中山大学附属第五医院健康管理中心；

【摘要】 目的探讨柚皮苷对高糖环境下MC3T3-E1细胞增殖、分化的影响及可能的分子机制。方法体外培养小鼠MC3T3-E1细胞,实验分5组:对照组、高糖组:25mmol/L葡萄糖、0.1μmol/L柚皮苷+25mmol/L葡萄糖、1μmol/L柚皮苷+25mmol/L葡萄糖、10μmol/L柚皮苷+25mmol/L葡萄糖。药物干预后,采用CCK-8法检测细胞增殖;qPCR法检测细胞IGF-1、Akt1、Runx2 mRNA的表达;Western Blot法检测细胞IGF-1、Akt1、ALP蛋白的表达。结果与对照组比,高糖组细胞增殖受到显著抑制(P<0.01)。不同浓度的柚皮苷组以剂量依赖方式明显改善高糖对细胞增殖的抑制(P<0.05或P<0.01)。qPCR结果显示,高糖组显著下调Akt1、Runx2 mRNA、IGF-1 mRNA的表达(P <0.01),而柚皮苷干预组IGF-1、Akt1、Runx2 mRNA的表达均上调。Western Blot结果显示,高糖组明显下调IGF-1、ALP、Akt1蛋白的表达(P <0.01或P <0.05),而柚皮苷干预组明显上调IGF-1、Akt1、ALP蛋白的表达(P<0.05或P<0.01)。结论柚皮苷在高糖环境下明显促进了MC3T3-E1细胞的增殖,并可能通过刺激IGF-1表达,激活Akt信号通路,进一步激活下游因子Runx2,最终促进ALP的表达,发挥促进骨形成的作用。更多还原

【Abstract】 objective To explore the effect of naringin on proliferation and differentiation of MC3 T3-E1 cells under high glucose environment and its possible molecular mechanism.Methods In vitro culture of mouse MC3 T3-E1 cells, The experiment is divided into 5 groups:The normal control group、the high glucose group: 25 mmol/L glucose、0.1 umol/L naringin +25 mmol/L glucose、1 umol/L naringin +25 mmol/L glucose、10 umol/L naringin +25 mmol/L glucose.After drug intervention, use CCK-8 to test cell proliferation; use qPCR method to test the mRNA expressions of IGF-1,Akt1 and Runx2; and use Western Blot method to test the protein expressions of IGF-1,Akt1 and ALP.Results Compared with the normal control group,Cell proliferation was significantly inhibited in the high glucose group( P < 0.01);Different concentrations of naringin significantly improved the inhibition of cell proliferation by high glucose in dose-dependent manner( P < 0.05 或 P < 0.01).qPCR results show,The expression of Akt1,Runx2 and IGF-1 mRNA was significantly decreased in the high-glucose group( P < 0.01),Different concentrations of naringin significantly up-regulated IGF-1,Akt1 and Runx2 mRNA expression in dose-dependent manner and with prolonged intervention time.Western Blot results show that,The expression of IGF-1, ALP and Akt1 proteins was significantly decreased in the high-glucose group( P < 0.01 或 P < 0.05),The expression of IGF-1, Akt1 and ALP protein was significantly up-regulated in different concentrations of naringinin the intervention group(P<0.05 或 P<0.01).Conclusions This experiment indicate that Naingin significantly promotes the MC3 T3-E1 cell proliferation in high glucose environment. And Its mechanism of action may stimulate the expression of IGF-1 to activate signal path of Akt, and further activate the downstream factor Runx2, and finally promote the expression of ALP, thus promoting the bone formation.更多还原