促红细胞生成素对肾间质纤维化大鼠肾脏保护作用的研究

发布时间：2018-05-30 12:47

本文选题：UUO大鼠 + 肾间质纤维化　；参考：《山西医科大学》2012年硕士论文

【摘要】：目的：研究幼年大鼠单侧输尿管梗阻模型（UUO模型）所制备的患侧肾间质纤维化中，给予重组人促红细胞生成素（rh-EPO）干预后，，α-平滑肌动蛋白（α-SMA）、转化生长因子-β1（TGF-β1）及血管内皮生长（VEGF）的表达趋势及变化情况，并探讨其对大鼠肾间质纤维化的影响及可能的作用机制。方法：采用单侧输尿管结扎造成大鼠患侧肾间质纤维化，2-3周龄SD大鼠72只，随机分成对照组即A组（仅分离输尿管，不结扎，腹腔注射生理盐水）、模型组即B组（造模，腹腔注射生理盐水）、模型干预组即C组（造模，腹腔注射rh-EPO100IU/Kg，3次/周）。于术后3、7、14天各组分别处死8只大鼠，取患侧肾做组织学检查，HE、Masson染色光镜下观察肾组织病理学变化，用免疫组化方法检测肾组织中α-SMA、TGF-β1、VEGF蛋白的表达及变化趋势，所得数据采用SPSS17.0统计学软件处理。结果:（1）病理变化：HE染色显示：A组大鼠肾组织结构未见明显变化。B组于术后3天出现肾小管轻度扩张，间质中少量单核细胞与淋巴细胞浸润；7天时可见肾小管扩张较前明显,偶见肾小管萎缩，同时可见肾小管上皮细胞变性、水肿、甚至坏死，肾间质炎症细胞浸润增多；14天上述病变继续加重，间质细胞及细胞外基质成分增多，纤维化程度较明显。与同期B组相比，C组大鼠肾小管扩张程度、间质中单核细胞及淋巴细胞浸润的程度有所减轻；与同期A组相比，C组大鼠肾脏病变程度仍较高。Masson染色显示：A组大鼠肾小管间质内未见明显阳染。随着梗阻时间的延长B组大鼠肾组织胶原纤维增加，染色面积逐渐增多。与同期B组比较，C组大鼠肾小管间质纤维化程度明显减轻，胶原纤维阳染面积减少(P㩳0.05)，但其程度仍高于同期A组(P㩳0.05)。（2）免疫组化结果显示:A组中α-SMA、TGF-β1在肾小管上皮细胞中有少量表达；于B组和C组中α-SMA、TGF-β1随着梗阻时间的延长表达水平逐渐上调，到14天表达最明显；C组中两种蛋白较B组同时间点的表达水平均明显下调(P㩳0.05)，但各时间点表达水平仍明显高于A组(P㩳0.05)；VEGF于肾小管上皮细胞及足突细胞中广泛表达，随着梗阻时间的延长，B组和C组中VEGF的表达均明显下调，但C组较B组各时间点的表达水平有所上调，但仍低于A组(P㩳0.05)。A组中三种蛋白的表达水平无明显变化，差异无统计学意义(P㩳0.05)。结论:研究显示α-SMA、TGF-β1蛋白的表达上调及VEGF蛋白的表达下调在肾间质纤维化的发展中具有重要的意义，用EPO干预后可能通过下调UUO模型大鼠肾组织中α-SMA、TGF-β1的蛋白表达，及上调VEGF蛋白的表达水平，从而具有延缓肾小管间质纤维化进展，保护肾脏的作用。
[Abstract]:Objective: to study the process of renal interstitial fibrosis induced by unilateral ureteral obstruction (UUO) in juvenile rats. After treated with recombinant human erythropoietin (rh-EPO), the expression trends and changes of 伪 -smooth muscle actin (伪 -SMA-TGF- 尾 1) and vascular endothelial growth factor (VEGF-1) were investigated, and the effects and possible mechanisms of 伪 -smooth muscle actin (TGF- 尾 _ 1) on renal interstitial fibrosis in rats were investigated. Methods: 72 SD rats of 2-3 weeks old with unilateral ureteral ligation were randomly divided into two groups: group A (isolated ureter, no ligation, intraperitoneal injection of normal saline), model group B (model group). Group C was treated with intraperitoneal injection of normal saline. The rats in group C were given intraperitoneal injection of rh-EPO100 IUP / KgN 3 times per week. On the 14th day after operation, 8 rats were killed in each group. The pathological changes of renal tissue were observed under light microscope with HE Masson staining, and the expression of 伪 -SMATGF- 尾 1 VEGF protein in renal tissue was detected by immunohistochemical method. The data were processed by SPSS17.0 software. Results the pathological changes showed that there was no obvious change in the renal tissue structure of the rats in group A. the renal tubule dilated slightly in group B on the 3rd day after operation, and the renal tubule dilatation was more obvious at 7 days after infiltration of a small amount of monocytes and lymphocytes in the interstitial tissue. Tubule atrophy was seen occasionally, and tubule epithelial cell degeneration, edema, and even necrosis, infiltration of renal interstitial inflammatory cells increased 14 days, interstitial cells and extracellular matrix increased, fibrosis degree was obvious. Compared with group B, the degree of renal tubule dilatation and the infiltration of monocytes and lymphocytes in interstitial tissue of rats in group C decreased. Compared with group A, the degree of renal lesion in group C was still higher. Masson staining showed that there was no obvious positive staining in tubulointerstitium of rats in group A. With the prolongation of obstruction time, the collagen fibers in renal tissue of group B increased and the staining area gradually increased. Compared with group B, the degree of renal tubulointerstitial fibrosis in group C was significantly reduced, and the area of positive staining of collagen fibers was decreased, but the degree was still higher than that in group A. The immunohistochemical results showed that 伪 -SMA-TGF- 尾 1 was expressed in a small amount in renal tubular epithelial cells. In group B and group C, the expression of 伪 -SMA-TGF- 尾 1 gradually increased with the prolongation of obstruction time. At the 14th day, the expression level of two proteins in group C was significantly lower than that in group B at the same time point, but the expression level of VEGF was still significantly higher than that of group A at the same time point, but the expression of VEGF in renal tubular epithelial cells and podocyte cells was significantly higher than that in group A. With the prolongation of the time of obstruction, the expression of VEGF in group B and group C was significantly down-regulated, but the expression level of VEGF in group C was up-regulated than that in group B, but still lower than that in group A (group A). There was no significant change in the expression of three proteins in group A. The difference was not statistically significant. Conclusion: the up-regulation of 伪 -SMA-TGF- 尾 1 protein and the down-regulation of VEGF protein may play an important role in the development of renal interstitial fibrosis. The expression of 伪 -SMA-TGF- 尾 1 protein may be down-regulated by the intervention of EPO. And up-regulate the expression of VEGF protein, which can delay the progression of tubulointerstitial fibrosis and protect the kidney.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363

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