人肾微血管内皮细胞转分化在移植肾间质纤维化形成中的作用研究
发布时间：2018-11-17 10:35
【摘要】：目的：探讨人肾微血管内皮细胞—间充质细胞转分化(Endothelial-mesenchymal transition, EndMT)在移植肾间质纤维化形成中的意义及相关机制。方法：通过对本院25例慢性移植肾失功能(Chronic allograft dysfunction,CAD)患者和25例正常人血生化指标、以及移植肾组织标本和正常肾组织标本进行、糖原(PAS)和马松三色(Masson)染色检查分析,观察两组研究对象肾功能、肾小管萎缩和肾小球塌陷及肾间质纤维化程度的差异。用免疫组织化学和间接免疫荧光双重染色方法检测两组肾组织标本中血管内皮细胞标志物——CD34和肌成纤维细胞标志物——αα-平滑肌细胞肌动蛋白(α-Smooth muscle actin, α-SMA)以及转化生长因子-β1 (Transforming growth factor-β1, TGF-β1)的表达和分布特点。进一步以原代人脐静脉内皮细胞(Human umbilical vein endothelial cell, HUVEC)为体外研究对象,予以TGF-β1 (5ng/m 1)分别作用0-72h,采用免疫蛋白印迹方法观察细胞中CD34和α-SMA的表达变化。结果：与正常人相比,CAD患者血清肌酐水平明显升高,且PAS和Masson三色染色结果显示CAD患者移植肾组织中出现明显肾小管萎缩、肾小球塌陷及肾间质纤维化改变;进一步半定量统计分析结果表明两者间存在统计学差异(P0.01)。免疫组织化学及间接免疫荧光染色结果表明,与正常组相比,CAD组中CD34阳性表达率降低而α-SMA及TGF-β1阳性表达率则显著升高。且间接免疫荧光双重染色研究发现CAD组中部分肾小球和间质微血管内皮细胞呈CD34和α-SMA双重染色阳性。体外研究显示,TGF-β1作用于HUVEC细胞后,随着时间延长,CD34表达逐渐降低而(α-SMA表达逐渐增多,与对照组相比,具有统计学差(P0.01)。结论：人肾微血管内皮细胞可能在TGF-β1介导下通过发生EndMT现象进而在移植肾间质纤维化形成中起重要作用。
[Abstract]:Aim: to investigate the significance and mechanism of human renal microvascular endothelial cell-mesenchymal cell transdifferentiation (Endothelial-mesenchymal transition, EndMT) in the formation of renal interstitial fibrosis. Methods: the blood biochemical indexes of 25 patients with chronic renal allograft dysfunction (Chronic allograft dysfunction,CAD) and 25 normal subjects were studied. The difference of renal function, renal tubular atrophy, glomerular collapse and renal interstitial fibrosis between the two groups was observed by (PAS) and Ma Song trichrome (Masson) staining. Detection of vascular endothelial cell markers-CD34 and myofibroblast markers-伪-smooth muscle actin (伪-Smooth muscle actin,) in renal tissues by immunohistochemistry and indirect immunofluorescence double staining The expression and distribution of 伪-SMA and transforming growth factor- 尾 1 (Transforming growth factor- 尾 1, TGF- 尾 1. Furthermore, the primary human umbilical vein endothelial cells (Human umbilical vein endothelial cell, HUVEC) were treated with TGF- 尾 1 (5ng/m 1) for 0-72 h in vitro. The expression of CD34 and 伪 SMA were observed by Western blot. Results: the serum creatinine level in CAD patients was significantly higher than that in normal controls. The results of PAS and Masson trichromatic staining showed that renal tubular atrophy, glomerular collapse and interstitial fibrosis were observed in CAD patients. Further semi-quantitative statistical analysis showed that there was a statistical difference between the two (P0.01). The results of immunohistochemistry and indirect immunofluorescence staining showed that the positive expression rate of CD34 in CAD group was lower than that in normal group, but the positive rate of 伪-SMA and TGF- 尾 1 was significantly increased in CAD group. Indirect immunofluorescence double staining showed that some glomerular and interstitial microvascular endothelial cells in CAD group were double stained with CD34 and 伪-SMA. In vitro studies showed that after TGF- 尾 1 was treated with HUVEC cells, the expression of CD34 decreased gradually and the expression of 伪-SMA increased with the prolongation of time. Compared with the control group, the expression of 伪-SMA was significantly lower than that of the control group (P0.01). Conclusion: human renal microvascular endothelial cells may play an important role in the formation of renal interstitial fibrosis by EndMT phenomenon mediated by TGF- 尾 1.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R692
[Abstract]:Aim: to investigate the significance and mechanism of human renal microvascular endothelial cell-mesenchymal cell transdifferentiation (Endothelial-mesenchymal transition, EndMT) in the formation of renal interstitial fibrosis. Methods: the blood biochemical indexes of 25 patients with chronic renal allograft dysfunction (Chronic allograft dysfunction,CAD) and 25 normal subjects were studied. The difference of renal function, renal tubular atrophy, glomerular collapse and renal interstitial fibrosis between the two groups was observed by (PAS) and Ma Song trichrome (Masson) staining. Detection of vascular endothelial cell markers-CD34 and myofibroblast markers-伪-smooth muscle actin (伪-Smooth muscle actin,) in renal tissues by immunohistochemistry and indirect immunofluorescence double staining The expression and distribution of 伪-SMA and transforming growth factor- 尾 1 (Transforming growth factor- 尾 1, TGF- 尾 1. Furthermore, the primary human umbilical vein endothelial cells (Human umbilical vein endothelial cell, HUVEC) were treated with TGF- 尾 1 (5ng/m 1) for 0-72 h in vitro. The expression of CD34 and 伪 SMA were observed by Western blot. Results: the serum creatinine level in CAD patients was significantly higher than that in normal controls. The results of PAS and Masson trichromatic staining showed that renal tubular atrophy, glomerular collapse and interstitial fibrosis were observed in CAD patients. Further semi-quantitative statistical analysis showed that there was a statistical difference between the two (P0.01). The results of immunohistochemistry and indirect immunofluorescence staining showed that the positive expression rate of CD34 in CAD group was lower than that in normal group, but the positive rate of 伪-SMA and TGF- 尾 1 was significantly increased in CAD group. Indirect immunofluorescence double staining showed that some glomerular and interstitial microvascular endothelial cells in CAD group were double stained with CD34 and 伪-SMA. In vitro studies showed that after TGF- 尾 1 was treated with HUVEC cells, the expression of CD34 decreased gradually and the expression of 伪-SMA increased with the prolongation of time. Compared with the control group, the expression of 伪-SMA was significantly lower than that of the control group (P0.01). Conclusion: human renal microvascular endothelial cells may play an important role in the formation of renal interstitial fibrosis by EndMT phenomenon mediated by TGF- 尾 1.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R692
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