CAJ | 학술논문

目的探讨转化生长因子β1(TGF-β1)的RNA干扰质粒(shRNA-TGF-β1)对大鼠移植肾上皮细胞转分化的作用.方法将受体大鼠分为4组:假手术组、质粒组、空质粒组、单纯移植组;建立大鼠肾移植纤维化加快模型,以流体力学为基础的肾脏基因转染方法转染质粒;分1、2、3个月收集肾脏标本;逆转录-PCR及Western印迹法检测TGF-β1、上皮钙黏蛋白(E-cadherin)的mRNA及蛋白表达;HE染色观察肾脏的病理变化;上皮钙黏蛋白及α肌动蛋白(α-SMA)免疫组化标记肾小管上皮细胞及成纤维母细胞.结果质粒组TGF-β1在3个检测时间段表达均受到抑制,明显低于空质粒组、单纯移植组[第3个月时TGF-β1 mRNA分别为0.73±0.08比0.92±0.07、0.95±0.04(均P<0.01)];质粒组上皮钙黏蛋白表达高于空质粒组、单纯移植组[第3个月时上皮钙黏蛋白mRNA分别为0.39±0.11比0.15±0.07、0.17±0.06(均P<0.01);上皮钙黏蛋白蛋白分别为0.38±0.08比0.15±0.07、0.15±0.07(均P<0.01)];质粒组肾小管上皮细胞多于空质粒组、单纯移植组,成纤维母细胞少于空质粒组、单纯移植组;病理染色显示质粒组慢性炎症较轻,纤维母细胞较少,细胞外基质沉积较轻.结论质粒shRNA-TGF-β1能抑制移植肾肾小管上皮细胞转分化,其机制可能是抑制TGF-β1的表达、上调上皮钙黏蛋白的表达.
목적 탐토전화생장인자β1(TGF-β1)적RNA간우질립(shRNA-TGF-β1)대대서이식신상피세포전분화적작용.방법 장수체대서분위4조:가수술조、질립조、공질립조、단순이식조;건립대서신이식섬유화가쾌모형,이류체역학위기출적신장기인전염방법전염질립;분1、2、3개월수집신장표본;역전록-PCR급Western인적법검측TGF-β1、상피개점단백(E-cadherin)적mRNA급단백표체;HE염색관찰신장적병리변화;상피개점단백급α기동단백(α-SMA)면역조화표기신소관상피세포급성섬유모세포.결과 질립조TGF-β1재3개검측시간단표체균수도억제,명현저우공질립조、단순이식조[제3개월시TGF-β1 mRNA분별위0.73±0.08비0.92±0.07、0.95±0.04(균P<0.01)];질립조상피개점단백표체고우공질립조、단순이식조[제3개월시상피개점단백mRNA분별위0.39±0.11비0.15±0.07、0.17±0.06(균P<0.01);상피개점단백단백분별위0.38±0.08비0.15±0.07、0.15±0.07(균P<0.01)];질립조신소관상피세포다우공질립조、단순이식조,성섬유모세포소우공질립조、단순이식조;병리염색현시질립조만성염증교경,섬유모세포교소,세포외기질침적교경.결론 질립shRNA-TGF-β1능억제이식신신소관상피세포전분화,기궤제가능시억제TGF-β1적표체、상조상피개점단백적표체.
Objective To investigate the effects of shRNA-transforming growth factor(TGF)-β1 plasmid upon epithelial-myofibroblast transdifferentiation of renal allograft in rats. Methods Divided the Wistar rats into 4 groups: Group J (sham-operated group), T (plasmid group), H (vacant plasmid group) and Y (simply transplantation group). The SD to Wistar rat transplant kidney-sclerosis accelerated model was constructed and transfected with the plasmid based on hydromechanics. Transplanted kidneys were collected at Months 1, 2 and 3 post-transplantation. The gene transcriptional levels of TGF-β1 and E-cadherin were detected by RT-PCR and the protein variation of E-cadherin was examined by Western blotting. The pathological changes and infiltrated inflammatory cells were assessed by HE staining and the immunohistochemical staining of E-cadherin and α-SMA used to label epithelial cells and fibroblast in order to exhibit cell transdifferentiation. Results Compared with Group H and Y, the mRNA transcription of TGF-β1 was obviously inhibited in the Group T: at Month 3, the TGF-β1 mRNA of Group T is 0.73 ± 0.08, significantly lower than Group H and Y (0.92±0.07 and 0.95±0.04, both P < 0.01) ; the expression of E-cadherin was maintained at a high level: at the Month 3, the E-cadherin mRNA of Group T is 0.39±0.11, significantly higher than Group H and Y (0.15±0.07, and 0.17±0.06, both P < 0.01); the E-cadherin protein of Group T is 0.38±0.08, significantly higher than group H and Y (0.15±0.07 and 0.15±0.07, both P < 0.01) ; epithelial cells were much more and fibroblast was much less than that of Group H and Y; there were also less infiltrated chronic inflammatory cells and extracellular matrix deposition in the Group T. Conclusion The shRNA-TGF-β1 plasmid can inhibit the epithelial-myofibroblast transdifferentiation of renal allograft in rats. The mechanisms may be associated with its effects of down-regulating TGF-β1 and up-regulating E-cadherin.