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

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-β1)、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-β1、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-β1、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表达和下调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。

Effect of bone morphogenetic protein-7 on advanced glycation end products induced epithelial-to-mesenchymal transition of rat peritoneal mesothelial cells

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-β1 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 α-SMA,collagen I,TGF-β1,VEGF and protein expression of α-SMA,TGF-β1 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 I.