VEGF-A、MCP-1及RANTES在血液透析患者自体动静脉内瘘狭窄部位表达的研究

发布时间：2018-01-19 03:26

本文关键词： 自体动静脉内瘘内膜增生血管内皮生长因子-A 单核细胞趋化蛋白-1 受激活调节正常T细胞表达和分泌　出处：《河北医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:自体动静脉内瘘(arteriovenous fistula,AVF)是血液透析患者首选的血管通路。但是AVF失功率较高。近吻合口静脉内膜增生,导致血管狭窄是AVF失功的最主要原因,在组织学上表现为血管平滑肌细胞的迁移和增殖,新生血管形成以及细胞外基质的过度沉积。尽管确切发病机制尚未清楚,但是实验和临床研究发现,多种细胞因子参与这一过程。血管内皮生长因子-A(Vascular endothelial growth factor-A,VEGF-A)可以促进内皮细胞和平滑肌细胞的增殖、迁移,同时能够促进单核细胞、巨噬细胞聚集。尤其是在血管重塑和再狭窄中发挥重要作用。炎症在AVF狭窄中起到重要作用,而单核细胞趋化蛋白-1(Monocyte chemoattractant protein-1,MCP-1)可以促进单核、巨噬细胞浸润到炎症部位。还能够活化并且促进平滑肌细胞向内膜的迁移与增殖,此外,它还可调节血管生成,并且在血管重塑和炎症性疾病中起到重要作用。受激活调节正常T细胞表达和分泌因子(Regulated upon activation,normal T-cell expressed and secreted,RANTES)作为一种趋化因子可以活化炎症细胞,并趋化炎症细胞向损伤和炎症部位聚集。尤其在促进血管生成中起到非常重要作用。本实验通过检测AVF静脉标本中VEGF-A、MCP-1和RANTES表达水平的变化,探讨VEGF-A、MCP-1以及RANTES与AVF内膜增生的关系。方法:选取28名已行规律血液透析治疗的终末期肾脏病患者,分为实验组和对照组。其中实验组14名为因近吻合口静脉狭窄导致AVF失功来我科行AVF重建术的患者。对照组则为拟初次行AVF成形术的14名患者。实验组患者行AVF重建术时,切取废弃的完整增生肥厚的静脉组织约1cm作为实验组;对照组患者初次行AVF端侧吻合术,将头静脉结扎切断后,于近心端切取0.5-1cm的完整静脉作为对照组。各组标本分别进行HE和免疫组织化学染色,并于光镜下观察。所得实验数据均采用SPSS 21.0统计软件进行分析,结果用均数±标准差(x±s)表示,p0.05认为差异具有统计学意义。结果:1患者临床基本统计学资料对2组患者年龄、性别及肾衰竭原发病构成比进行统计学分析,差异均无统计学意义(p0.1)。2 HE染色光镜下观察发现实验组失功AVF的静脉内膜明显增厚,并有大量炎细胞浸润和新生血管形成,对照组静脉内膜未见明显增生肥厚,无明显炎细胞浸润及新生血管形成,对实验组和对照组的内膜/中膜厚度比和内膜/中膜面积比进行了比较,差异具有统计学意义(p均0.001)。3免疫组织化学染色3.1 VEGF-A免疫组织化学染色VEGF-A表达主要位于胞浆中。在实验组中以强阳性表达为主,主要位于中膜和内膜;而对照组静脉VEGF-A表达呈弱阳性,主要位于内膜和中膜,其中内膜较明显。实验组VEGF-A的表达水平明显高于对照组,差异具有统计学意义(p0.01)。3.2 MCP-1免疫组织化学染色MCP-1主要表达于胞浆中。实验组以强阳性表达为主,位于静脉内膜和中膜。而对照组MCP-1表达以弱阳性为主,位于静脉内膜和中膜,以中膜更明显。实验组MCP-1的表达水平明显高于对照组,差异具有统计学意义(p0.01)。3.3 RANTES免疫组化染色RANTES主要表达于胞浆。实验组RANTES表达以强阳性为主,主要在内膜和中膜。对照组RANTES呈弱阳性表达,位于内膜和中膜。实验组RANTES的表达水平明显高于对照组,差异具有统计学意义(p0.01)。3.4免疫组织化学染色的阴性对照:阴性对照组内膜和中膜均未见阳性染色,血管外膜呈棕黄色,但是未见阳性细胞,考虑为非特异性染色。结论:1 VEGF-A、MCP-1及RANTES的高表达与静脉内膜增生密切相关,可能成为未来干预的靶点。2 VEGF-A可能通过介导MCP-1的表达,继而介导RANTES的表达,在AVF内膜增生中发挥作用。3 VEGF-A也可能直接介导MCP-1和RANTES的表达,在AVF内膜增生中发挥作用。
[Abstract]:Objective: autologous arteriovenous fistula (arteriovenous fistula AVF) is the preferred vascular access in hemodialysis patients. But the AVF loss of high power. Nearly kiss intimal hyperplasia stoma, lead to vascular stenosis is the main cause of AVF dysfunction, in histologically vascular smooth muscle cell migration and proliferation, angiogenesis and excessive deposition of extracellular matrix. Although the exact pathogenesis is not clear, but the experimental and clinical study found that many cytokines involved in this process. Vascular endothelial growth factor -A (Vascular endothelial growth factor-A, VEGF-A) can promote endothelial cell and smooth muscle cell proliferation, migration, and can promote monocyte macrophage accumulation. Especially play an important role in vascular remodeling and restenosis. Inflammation plays an important role in AVF stenosis, and monocyte chemoattractant protein -1 (Monocyte Ch Emoattractant protein-1, MCP-1) can promote monocyte, macrophage infiltration into inflammatory sites. Also can activate and promote smooth muscle cell migration and proliferation of the endometrium, in addition, it can also regulate angiogenesis, and play an important role in vascular remodeling and inflammatory diseases. Regulated upon activation normal T cell expressed and secreted (Regulated upon activation, normal T-cell expressed and secreted, RANTES) is a chemokine activation of inflammatory cells, and chemotactic aggregation of inflammatory cells to injury and inflammation in the body. Especially in the promotion play an important role in angiogenesis. The detection of AVF MCP-1 and VEGF-A in venous samples, the change of the expression of RANTES to investigate, VEGF-A, MCP-1 and AVF between RANTES and endometrial hyperplasia. Methods: 28 patients who underwent hemodialysis treatment in patients with end-stage renal disease, divided into The experimental group and control group. The experimental group was 14 for anastomotic vein stenosis resulting in AVF loss to our department for reconstruction of AVF patients. The control group was intended to initial AVF plasty in 14 patients. The experimental group underwent AVF reconstruction, vein cut complete hypertrophy from waste about the 1cm as the experimental group; the control group of patients undergoing primary AVF end to side anastomosis, the cephalic vein ligation after complete venous 0.5-1cm cut as control group at the proximal end. All specimens were HE and immunohistochemical staining, and observed under light microscope. The experimental data were treated by SPSS 21 statistical software for analysis, the standard deviation (x + s) said, P0.05 considered statistically significant. Results: 1 patients with clinical basic demographic data of 2 groups of patients age, proportion for statistical analysis of gender and renal failure of primary disease, the differences were No statistical significance (P0.1) found that the experimental group loss AVF venous intima thickening obviously were observed under light microscope..2 HE staining, and a large number of inflammatory cell infiltration and neovascularization, the control group had no obvious intimal hypertrophy, no obvious inflammatory cell infiltration and neovascularization of the experimental group and control group endometrium / in the film thickness and the ratio of intima / media area ratio were compared, the difference was statistically significant (P 0.001).3 immunohistochemistry 3.1 VEGF-A immunohistochemical staining of VEGF-A expression was mainly located in the cytoplasm. With strong positive expression in the experimental group, mainly located in the membrane and the membrane; and the control group the expression of VEGF-A was weakly positive, mainly located in the intima and media, the endometrium is obvious. The expression level of VEGF-A in the experimental group was significantly higher than the control group, the difference was statistically significant (P0.01).3.2 MCP-1 immunohistochemistry MCP -1 was mainly expressed in the cytoplasm. The experimental group with strong positive expression mainly located in the intima and media of vein. While the control group with weak positive MCP-1 expression mainly located in venous intima and media, in the film is more obvious. The expression level of MCP-1 in the experimental group was significantly higher than the control group, the difference has statistical significance (P0.01.3.3 RANTES) immunohistochemical staining of RANTES was mainly expressed in the cytoplasm. RANTES expression in the experimental group with strong positive, mainly in the intima and the control group. The expression of RANTES was weakly positive, located in the intima and media. The expression level of RANTES in the experimental group was significantly higher than the control group, the difference was statistically significant (P0.01) immune.3.4 histochemical staining of the negative control: negative control group, intima and adventitia showed no positive staining was brown, but no positive staining cells, considered nonspecific. Conclusion: 1 VEGF-A, high expression of MCP-1 and RANTES and static Vein intimal hyperplasia is closely related to the expression, may become targets of.2 VEGF-A future intervention may be mediated by MCP-1, then the expression of RANTES mediated by.3 VEGF-A AVF, play a role in endometrial hyperplasia may also direct expression mediated by MCP-1 and RANTES, AVF play a role in neointimal hyperplasia.

【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R459.5

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