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目的 观察转化生长因子beta1（TGF-β1）反义RNA对系膜细胞基质合成的影响。方法 构建含TGF-β1反义RNA的重组腺病毒,将重组腺病毒转染系膜细胞,用Northern blot检测转染系膜细胞TGF-β1及ColⅣmRNA的含量,用免疫组化半定量分析转染的系膜细胞中TGF-β1、纤连蛋白（FN）及胶原（Col）Ⅳ蛋白水平,并与无转染的系膜细胞对照组比较其表达的变化。结果 构建了含TGF-β1反义RNA的重组腺病毒。重组反义TGF-β1腺病毒转染系膜细胞后,与对照组相比,在第24hTGF-β1及ColⅣ的mRNA无明显抑制,在48h TGF-β1及ColⅣmRNA抑制率分别为22.5％,18.2％;72h TGF-β1及ColⅣmRNA抑制率分别为29.5％,27.3％。免疫组织化学半定量结果显示:与对照组相比,在48h始转染系膜细胞TGF-β1、FN及ColⅣ蛋白含量开始下降,48hTGF-β1、FN及ColⅣ蛋白抑制率分别为16.5％,18.2％及14.6％;72hTGF-β1、FN及ColⅣ蛋白抑制率分别为23.5％,27.3％及26.8％。结论 重组反义TGF-β1可抑制系膜细胞合成细胞外基质,在肾小球肾炎及肾小球硬化的研究及治疗中有潜在的应用价值。
Objective To observe the effects of transforming growth factor-beta1 (TGF-β1) antisense RNA on mesangial matrix synthesis. Methods The recombinant adenovirus containing TGF-β1 antisense RNA was constructed and the recombinant adenovirus was transfected into mesangial cells. The content of TGF-β1 and ColⅣ mRNA in transfected mesangial cells was detected by Northern blot. The expression of TGF-β1 and ColⅣ mRNA was detected by semi-quantitative immunohistochemistry TGF-β1, fibronectin (FN) and collagen Ⅳ protein in mesangial cells were detected by Western blot and compared with that of non-transfected mesangial cell control group. Results A recombinant adenovirus containing TGF-β1 antisense RNA was constructed. Compared with the control group, the mRNA expression of TGF-β1 and ColⅣ in the transfected mesangial cells of recombinant antisense TGF-β1 group was not significantly inhibited at 24h, and the inhibitory rates of TGF-β1 and ColⅣ mRNA were 22.5%, 18.2% ; 72h TGF-β1 and Col Ⅳ mRNA inhibition rates were 29.5%, 27.3%. Semi-quantitative immunohistochemistry results showed that compared with the control group, the protein levels of TGF-β1, FN and ColⅣ began to decrease 48 h after transfection, and the inhibitory rates of 48 h TGF-β1, FN and ColⅣ were 16.5% and 18.2 % And 14.6% respectively. The inhibition rates of 72hTGF-β1, FN and ColⅣ protein were 23.5%, 27.3% and 26.8% respectively. Conclusion Recombinant antisense TGF-β1 can inhibit mesangial cells to synthesize extracellular matrix, which has potential value in the study and treatment of glomerulonephritis and glomerulosclerosis.