淡豆豉异黄酮通过影响Sox2对C3H10T1/2细胞增殖和分化的影响

Influence of fermented soybean isoflavone on C3H10T1/2 cell proliferation and differentiation via Sox2

目的探讨淡豆豉异黄酮通过影响Sox2对C3H10T1/2细胞增殖和分化的影响。方法选取间充质干细胞株C3H10T1/2为研究对象,不同浓度淡豆豉异黄酮(0、10、100、1 000μg/L)处理C3H10T1/2细胞,并同步联合成骨诱导液诱导成骨细胞分化,采用四甲基偶氮唑蓝(MTT)实验检测C3H10T1/2细胞增殖情况,采用碱性磷酸酶(ALP)活性实验鉴定C3H10T1/2细胞成骨分化情况,实时荧光定量PCR(q RT-PCR)法及蛋白印迹法检测C3H10T1/2细胞中Sox2 m RNA及蛋白表达情况。构建p EGFP-N1-Sox2过表达载体,利用脂质体2000向C3H10T1/2细胞转染重组质粒,分析Sox2过表达对C3H10T1/2细胞增殖和成骨分化的影响。结果 MTT检测结果可见,10、100、1 000μg/L淡豆豉异黄酮可明显抑制C3H10T1/2细胞增殖,且呈现一定时间、剂量依赖性(P<0.05);10、100、1 000μg/L淡豆豉异黄酮可明显促进C3H10T1/2细胞成骨分化,且呈一定剂量依赖性(P<0.05);10、100、1 000μg/L淡豆豉异黄酮可明显抑制C3H10T1/2细胞中Sox2 m RNA及蛋白表达,且呈一定剂量依赖性(P<0.05);Sox2过表达可明显促进C3H10T1/2细胞增殖,抑制成骨细胞分化。结论淡豆豉异黄酮可能是通过下调Sox2表达,发挥抑制C3H10T1/2细胞增殖,促进成骨细胞分化作用。

Objective To explore the influence of fermented soybean isoflavone on C3 H10 T1/2 cell proliferation and osteogenic differentiation via Sox2.Methods Mesenchymal stem cell line C3 H10 T1/2 was treated with different concentrations of fermented soybean isoflavone（0,10,100,1 000 μg/L）,and induced to osteogenic differentiation by treating with simultaneous combined osteogenic induction fluid.C3 H10 T1/2 cell proliferation was detected by methyl blue tetrazolium（MTT）assay;C3 H10 T1/2 cells osteogenic differentiation was determined by alkaline phosphatase（ALP）activity test.Sox2 m RNA and protein expressions in C3 H10 T1/2 cells were detected by real time fluorescence quantitative PCR（q RT-PCR）and Western blotting method,respectively.PEGFP-N1-Sox2 overexpression vector was constructed;recombinant plasmid was transfected into C3 H10 T1/2 cell by using liposomes 2000.Results MTT results showed that fermented soybean isoflavone could obviously inhibit C3 H10 T1/2 cells proliferation,and showed a time and dose dependent manner（P〈0.05）.Fermented soybean isoflavone could significantly promote C3 H10 T1/2 cells osteogenic differentiation in a dose dependent manner（P〈0.05）.Fermented soybean isoflavone could obviously inhibit Sox2 m RNA and protein expressions in C3 H10 T1/2 cells,and showed a dose dependent manner（P〈0.05）.Sox2 overexpression could promote C3 H10 T1/2 cells proliferation and inhibit osteoblasts differentiation.Conclusion Fermented soybean isoflavone may play a role in inhibiting C3 H10 T1/2 cell proliferation and promoting osteoblast differentiation by down-regulating Sox2 expression.