CAJ | 학술논문

目的：探讨前列环素类似物贝前列素钠（BPS）对高糖条件下大鼠系膜细胞细胞外基质代谢的影响及其可能的机制。方法将实验分为对照（NG）组、高糖（HG）组和高糖联合不同浓度BPS组（0.5、1、2、5μmol/L）。体外培养系膜细胞，收集24、48 h时细胞培养上清液，Elisa法测转化生长因子β1（TGFβ1）、纤维连接蛋白（FN）及基质金属蛋白酶-2（MMP-2）蛋白的表达；提取细胞总蛋白，免疫印迹法测Smad3磷酸化蛋白的表达。结果与NG组相比，HG培养24及48 h细胞TGFβ1、FN蛋白表达均显著增加（P<0.01），MMP-2蛋白表达量显著下降（P<0.01）；细胞培养24、48 h后，与HG组相比，HG+1μmol/L BPS组、HG+2μmol/L BPS组及HG+5μmol/LBPS组细胞TGFβ1蛋白表达量均显著降低（P<0.01）；HG+2μmol/L BPS组、HG+5μmol/L BPS组细胞FN蛋白表达量均显著降低（P<0.01）；HG+2μmol/L BPS组及HG+5μmol/L BPS组细胞MMP-2蛋白表达量均显著增加（P<0.05）；HG组细胞Smad3磷酸化蛋白表达较NG组显著增加（P<0.01）；与HG组相比，HG+2μmol/L BPS组与HG+5μmol/L BPS组细胞Smad3蛋白磷酸化水平显著下降（P<0.05）。结论BPS可能通过抑制TGFβ1/Smad3通路活性对高糖条件下系膜细胞ECM代谢起到了保护性的调节作用。
목적：탐토전렬배소유사물패전렬소납（BPS）대고당조건하대서계막세포세포외기질대사적영향급기가능적궤제。방법장실험분위대조（NG）조、고당（HG）조화고당연합불동농도BPS조（0.5、1、2、5μmol/L）。체외배양계막세포，수집24、48 h시세포배양상청액，Elisa법측전화생장인자β1（TGFβ1）、섬유련접단백（FN）급기질금속단백매-2（MMP-2）단백적표체；제취세포총단백，면역인적법측Smad3린산화단백적표체。결과여NG조상비，HG배양24급48 h세포TGFβ1、FN단백표체균현저증가（P<0.01），MMP-2단백표체량현저하강（P<0.01）；세포배양24、48 h후，여HG조상비，HG+1μmol/L BPS조、HG+2μmol/L BPS조급HG+5μmol/LBPS조세포TGFβ1단백표체량균현저강저（P<0.01）；HG+2μmol/L BPS조、HG+5μmol/L BPS조세포FN단백표체량균현저강저（P<0.01）；HG+2μmol/L BPS조급HG+5μmol/L BPS조세포MMP-2단백표체량균현저증가（P<0.05）；HG조세포Smad3린산화단백표체교NG조현저증가（P<0.01）；여HG조상비，HG+2μmol/L BPS조여HG+5μmol/L BPS조세포Smad3단백린산화수평현저하강（P<0.05）。결론BPS가능통과억제TGFβ1/Smad3통로활성대고당조건하계막세포ECM대사기도료보호성적조절작용。
Objective To explore effects of beraprost sodium (BPS) on the metabolism of extracellular matrix (ECM) in rat mesangial cells cultured in the presence of high glucose and the possible mechanism. Methods Rat mesangial cells were cultured in the presence of high glucose with or without BPS for 24 or 48 h. The levels of transforming growth factorβ1 (TGFβ1), fibronectin (FN) and matrix metalloproteinase-2 (MMP-2) protein in the culture supernatants were measured by enzyme-linked immunosorbent assay, and photoshop-Smad3 was detected by Western blotting. Results Compared with the cells in normal glucose, the cells cultured in the presence of high glucose for 24 and 48 h showed significantly increased TGFβ1 and FN protein expression and lowered MMP-2 protein expression (P<0.01). Compared with the cells cultured in high glucose, BPS exposure at the concentration of 1, 2, and 5 μmol/L for 24 and 48 h significantly lowered TGFβ1 protein expression (P<0.01), and at 2 and 5 μmol/L, BPS significantly decreased FN protein expression and increased MMP-2 protein expression in high glucose-induced cells (P<0.05). High glucose exposure also significantly increased the expression phosphorylated Smad3 (P<0.01), which was lowered by BPS treatment at 2 and 5μmol/L (P<0.01). Conclusion BPS can regulate ECM metabolism in rat mesangial cells cultured in high glucose by inhibiting TGFβ1/Smad3 pathway, suggesting the beneficial effects of BPS in preventing and treating diabetic nephropathy.