Xia Yu-guo, Wu Xiao-hou, Yin Zhi-kang, Luo Chun-li, Zhang Jia-mo, Cheng Hong-lin
Department of Urology, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China.
Zhonghua Yi Xue Za Zhi. 2009 Dec 1;89(44):3151-5.
To investigate the effects of shRNA-transforming growth factor (TGF)-beta1 plasmid upon epithelial-myofibroblast transdifferentiation of renal allograft in rats.
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-beta1 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 alpha-SMA used to label epithelial cells and fibroblast in order to exhibit cell transdifferentiation.
Compared with Group H and Y, the mRNA transcription of TGF-beta1 was obviously inhibited in the Group T: at Month 3, the TGF-beta1 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.
The shRNA-TGF-beta1 plasmid can inhibit the epithelial-myofibroblast transdifferentiation of renal allograft in rats. The mechanisms may be associated with its effects of down-regulating TGF-beta1 and up-regulating E-cadherin.
探讨shRNA-转化生长因子（TGF）-β1质粒对大鼠同种异体移植肾上皮-肌成纤维细胞转分化的影响。
将Wistar大鼠分为4组：J组（假手术组）、T组（质粒组）、H组（空质粒组）和Y组（单纯移植组）。构建SD大鼠至Wistar大鼠移植肾硬化加速模型，并采用流体力学方法进行质粒转染。于移植后1、2和3个月采集移植肾。通过RT-PCR检测TGF-β1和E-钙黏蛋白的基因转录水平，采用Western印迹法检测E-钙黏蛋白的蛋白变化。通过HE染色评估病理变化及炎性细胞浸润情况，采用E-钙黏蛋白和α-平滑肌肌动蛋白的免疫组化染色标记上皮细胞和成纤维细胞，以显示细胞转分化情况。
与H组和Y组相比，T组TGF-β1的mRNA转录明显受到抑制：在第3个月时，T组的TGF-β1 mRNA为0.73±0.08，显著低于H组和Y组（分别为0.92±0.07和0.95±0.04，P均<0.01）；E-钙黏蛋白的表达维持在较高水平：在第3个月时，T组的E-钙黏蛋白mRNA为0.39±0.11，显著高于H组和Y组（分别为0.15±0.07和0.17±0.06，P均<0.01）；T组的E-钙黏蛋白蛋白为0.38±0.08，显著高于H组和Y组（分别为0.15±0.07和0.15±0.07，P均<0.01）；上皮细胞较H组和Y组更多，而成纤维细胞较H组和Y组更少；T组的慢性炎性细胞浸润及细胞外基质沉积也更少。
shRNA-TGF-β1质粒可抑制大鼠同种异体移植肾上皮-肌成纤维细胞转分化。其机制可能与其下调TGF-β1及上调E-钙黏蛋白的作用有关。
