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目的观察洛伐他汀对人常染色体显性遗传性多囊肾病(ADPKD)囊肿衬里上皮细胞COLⅠα1基因启动子活性的影响。方法用FuGENE转染法将pCOLH0.27或pCOLH2.5重组体瞬时转染至ADPKD囊肿衬里上皮细胞,用不同浓度的洛伐他汀和香叶基香叶基焦磷酸(GGPP)处理转染细胞,24h后,用ELISA法测定细胞报告基因CAT表达量。以未加入药物孔的CAT值为1,计算出实验孔CAT的相对表达量。结果瞬时转染pCOLH0.27或pCOLH2.5的ADPKD囊肿衬里上皮细胞,经10或50μmol/L洛伐他汀处理24h后,pCOLH0.27与pCOLH2.5的活性均受明显抑制。10μmol/L洛伐他汀使pCOLH0.27与pCOLH2.5活性分别下降22%和26%(P<0.05),50μmol/L洛伐他汀使pCOLH0.27与pCOLH2.5活性分别下降37%和39%(P<0.01)。洛伐他汀对pCOLH0.27与pCOLH2.5活性的抑制水平相近。GGPP可逆转这一作用(P<0.01)。结论洛伐他汀对ADPKD囊肿衬里上皮细胞COLⅠα1基因启动子活性有明显抑制作用。洛伐他汀的这一作用可能与其抑制GGPP产生,从而阻断促纤维化细胞因子信号转导有关。
Objective To investigate the effect of lovastatin on the promoter activity of COLⅠα1 gene in cyst lining epithelial cells of autosomal dominant polycystic kidney disease (ADPKD). Methods The recombinant plasmid pCOLH0.27 or pCOLH2.5 was transiently transfected into the epithelial cells of ADPKD cysts by FuGENE transfection method. The transfected cells were treated with different concentrations of lovastatin and geranylgeranyl pyrophosphate (GGPP) for 24 h Afterwards, the CAT expression level of the cell reporter gene was measured by ELISA. With no added drug hole CAT value of 1, calculate the experimental hole CAT relative expression. Results After transient transfection of ADPKD cyst-lining epithelial cells with pCOLH0.27 or pCOLH2.5, the activities of pCOLH0.27 and pCOLH2.5 were significantly inhibited after 10 or 50μmol / L lovastatin treatment. The activity of pCOLH0.27 and pCOLH2.5 decreased by 22% and 26% (P <0.05) with 10μmol / L lovastatin, while the activity of pCOLH0.27 and pCOLH2.5 decreased by 37% and 39% respectively with 50μmol / L lovastatin. (P <0.01). Lovastatin had similar inhibitory effect on pCOLH0.27 and pCOLH2.5 activity. GGPP reversed this effect (P <0.01). Conclusion Lovastatin has a significant inhibitory effect on the promoter activity of COL Ⅰ α1 gene in the epithelial cells of ADPKD cysts. This role of lovastatin may be related to its inhibition of GGPP production, thereby blocking the signal transduction of pro-fibrotic cytokines.