骨形成蛋白-7对晚期糖基化终产物诱导大鼠腹膜间皮细胞上皮-间叶转化的影响

摘要

目的 探讨骨形成蛋白-7(BMP-7)对晚期糖基化终产物诱导大鼠腹膜间皮细胞上皮-间叶转化(EMT)的影响.方法 晚期糖基化终产物诱导发生上皮-间叶转化的体外培养大鼠腹膜间皮细胞,分别经含5 ng/mL及80 mmol/L晚期糖基化终产物的M199培养基和含10 ng/mL BMP-7及80 mmol/L晚期糖基化终产物的M199培养基培养48 h,以含80 mmol/L晚期糖基化终产物的M199培养基为对照,应用实时定量PCR法检测间皮细胞E-cadherin、α平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原(Collagen Ⅰ)、转化生长因子β1(TGF-31)、血管内皮生长因子(VEGF) mRNA的表达；应用Western印迹法检测E-cadherin、α-SMA蛋白的表达；应用ELISA法检测间皮细胞TGF-31、VEGF蛋白表达水平.结果 BMP-7作用后,上皮-间叶转化的大鼠腹膜间皮细胞E-cadherin mRNA和E-cadherin蛋白表达水平显著增加(P＜0.05).BMP-7作用后,上皮-间叶转化的大鼠腹膜间皮α-SMA、Collagen Ⅰ、TGF-31、VEGFmRNA和α-SMA、TGF-β、VEGF蛋白表达水平显著下降(P＜0.05).结论 BMP-7能上调上皮-间叶转化的大鼠腹膜间皮细胞E-cadherin表达和下凋α-SMA、Collagen Ⅰ、TGF-β、VEGF表达,BMP-7能逆转晚期糖基化终产物诱导大鼠腹膜间皮细胞EMT.%Objective To investigate the effect of bone morphogenetic protein-7 on advanced glycation end products induced epithelial-to-mesenchymal transition of rat peritoneal mesothelial cells. Methods Rat peritoneal mesothelial cells induced epithelial-to-mesenchymal transition by advanced glycation end products were exposed to culture medium containing different concentrations of bone morphogenetic protein-7 (0 ng/mL, 5 ng/mL, 10 ng/mL) and 80 mmol/L advanced glycation end products for 48 hours. Then the mRNA levels expression of E-cadherin, α-smooth muscle actin (α-SMA) , collagen I, TGF-β1 , VEGF and were determined with Real-Time PCR. The protein levels expression of E-cadherin and α-SMA were determined with Western blotting. The protein levels expression of TGF-βl and VEGF were determined with ELISA. Results The mRNA levels and protein expression of E-cadherin was significantly higher in BMP-7 (5 ng/mL, 10 ng/mL) group than that in advanced glycation end products group (P<0.05). The mRNA levels of a-SMA, collagen I, TGF-β1 , VEGF and protein expression of a-SMA, TGF-βl and VEGF was significantly lower in BMP-7 (5 ng/mL, 10 ng/mL) group than that in advanced glycation end products group (P <0.05). Conclusion BMP-7 can reverse advanced glycation end products induced epithelial-to-mesenchymal transition of rat peritoneal mesothelial cells, which is proved by acquirement of E-cadherin and loss of α-SMA and Collagen 1.