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目的探讨脂肪间充质干细胞(ADSC)经Wnt/β联蛋白(β-catenin)通路对肾小球系膜细胞增殖的影响。方法以大鼠ADSC的条件培养基作用于HBZY-1大鼠肾小球系膜细胞,通过流式细胞术分析细胞周期、RNA干扰、实时定量PCR和Western blot等技术分析HBZY-1细胞增殖能力、Wnt信号通路中相关基因和蛋白表达水平的变化。结果在ADSC条件培养基作用下,HBZY-1细胞增殖受到明显抑制,dickkopf WNT信号通路抑制物1(DKK1)mRNA表达水平明显升高,而纤连蛋白(fibronectin)和转化生长因子β1(TGF-β1)的mRNA表达显著降低,同时系膜细胞的β-catenin和Bcl-2蛋白的表达受到明显抑制。ADSC中DKK1 mRNA表达水平显著高于HBZY-1细胞,利用小干涉RNA敲低ADSC的DKK1表达后,其β-catenin蛋白的表达水平显著升高。将干扰后的ADSC条件培养基作用于HBZY-1细胞,可重新增加HBZY-1细胞Wnt信号通路中β-catenin和Bcl-2的蛋白水平。结论 Wnt/β-catenin信号通路可能参与ADSC对HBZY-1肾小球系膜细胞增殖的调控。
Objective To investigate the effect of adipose-derived mesenchymal stem cells (ADSC) on the proliferation of mesangial cells via Wnt / β-catenin pathway. Methods Rat mesangial cells were cultured with conditioned media of rat ADSCs. The cell cycle, RNAi, real-time PCR and Western blot were used to analyze the proliferation of HBZY-1 cells by flow cytometry , Wnt signaling pathway related genes and protein expression levels. Results The proliferation of HBZY-1 cells was significantly inhibited under the condition of ADSC conditioned medium, and the expression of DKK1 mRNA in dickkopf WNT pathway was significantly increased. However, the expression of fibronectin and transforming growth factor-β1 (TGF- β1) mRNA expression was significantly decreased, while mesangial cells β-catenin and Bcl-2 protein expression was significantly inhibited. The expression of DKK1 mRNA in ADSCs was significantly higher than that in HBZY-1 cells. The knockdown of ADK DKK1 expression by small interfering RNA significantly increased the expression of β-catenin protein. After interfering with ADSC conditioned medium in HBZY-1 cells, the protein level of β-catenin and Bcl-2 in Wnt signaling pathway of HBZY-1 cells was increased again. Conclusion Wnt / β-catenin signaling pathway may be involved in the regulation of ADSC on the proliferation of mesangial cells in HBZY-1.