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

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

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。

Objective To investigate the effect of bone morphogenetic protein-7 on advanced glycation end products induced epithelial-to-mesenehymal 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 mor- phogenetic protein-7 （0 ng/mL, 5 ng/mL, 10 ng/mL） and 80 mmol/L advanced glycation end prod- ucts for 48 hours. Then the mRNA levels expression of E-cadberin, c^-smooth muscle actin （ct-SMA） , collagen I, TGF-~31, VEGF and were determined with Real-Time PCR. The protein levels expression of E-cadherin and ot-SMA were determined with Western blotting. The protein levels expression of TGF-~I and VEGF were determined with ELISA. Results The mRNA levels and protein expression of E-eadherin was significantly higher in BMP-7 （5 ng/mL, 10 ng/mL） group than that in advanced glyeation end products group （P 〈0.05）. The mRNA levels of t^-SMA, collagen I, TGF-~I, VEGF and protein expression of cx-SMA, TGF-~I 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 perito- neal mesothelial cells, which is proved by acquirement of E-cadherin and loss of ot-SMA and Collagen I.