胶原结合血管内皮细胞生长因子抗小鼠肝纤维化作用研究

发布时间：2018-03-03 13:48

本文选题：血管内皮细胞生长因子　切入点：胶原结合区　出处：《南京大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：探讨胶原结合血管内皮细胞生长因子(Collagen-binding-domain vascular endothelial growth factor, CBD-VEGF)抗四氯化碳(CCh)诱导的小鼠肝纤维化作用及其可能机制。方法：BALB/c小鼠腹腔注射CCl4油剂(CCl4：橄榄油=1：5,5μl/g),每周2次,连续12周建立小鼠肝纤维化模型。将肝纤维化小鼠随机分为三组：正常对照组、生理盐水对照组和CBD-VEGF组。CBD-VEGF组肝脏局部注射50山CBD-VEGF (10μl),生理盐水对照组肝脏局部注射50/μl生理盐水。注射后4周及8周各组随机处死小鼠,采集外周血,分离血清,检测肝功能。同时,取出肝组织,进行HE和Masson染色观察肝组织形态和纤维化程度,免疫组织化学方法检测肝组织中CD31、α-平滑肌激动蛋白(a-SMA)及Ki-67的表达,TUNEL法检测肝细胞凋亡。结果CCh腹腔注射12周后,小鼠肝组织HE染色可见部分肝细胞肿胀、坏死,汇管区和小叶间有大量炎症细胞浸润,正常肝小叶结构明显紊乱。Masson染色见肝组织内纤维增生明显,肝组织被胶原纤维所分隔,形成假小叶,表明小鼠肝纤维化模型建立成功。CBD-VEGF注射后,肝功能检测结果显示,注射后4周,CBD-VEGF组ALT、AST水平高于NS对照组,而8周时两组ALT、AST水平无统计学差异。肝组织HE染色示,CBD-VEGF注射后4周和8周小鼠肝脏炎症程度明显轻于NS对照组。Masson染色表明CBD-VEGF组肝纤维化程度较NS对照组明显减轻。免疫组织化学表明CBD-VEGF组微血管数量明显多于NS对照组(p0.01),肝星状细胞活化程度低于NS对照组(p0.01),肝细胞增殖高于NS对照组(p0.01)。TUNEL染色表明CBD-VEGF组肝细胞凋亡明显较NS对照组轻(p0.05)。结论肝脏组织局部注射CBD-VEGF能够显著减轻CC14诱导的小鼠肝纤维化。CBD-VEGF的抗纤维化作用可能与其促进肝组织内毛细血管再生,促进肝细胞增殖,减少肝细胞凋亡,抑制肝星状细胞(hepatic stellate cells, HSCs)活化有关。
[Abstract]:Objective: to investigate the effect of collagen-binding-domain vascular endothelial growth factor (CBD-VEGF) on hepatic fibrosis induced by CCL _ 4 in mice and its possible mechanism. The mice with hepatic fibrosis were randomly divided into three groups: the normal control group, the control group, and the control group. Normal saline control group and CBD-VEGF group. CBD-VEGF group was given local injection of 50 mountain CBD-VEGF 10 渭 l in liver, and 50 / 渭 l normal saline was injected locally in normal saline control group. The mice were killed at random 4 and 8 weeks after injection, peripheral blood was collected and serum was isolated. At the same time, the liver tissue was taken out, and the morphology and fibrosis degree of liver tissue were observed by HE and Masson staining. Immunohistochemical method was used to detect the expression of CD31, 伪 -smooth muscle activin a-SMAand Ki-67 in liver tissue and Tunel method was used to detect the apoptosis of hepatocytes. Results after 12 weeks of intraperitoneal injection of CCh, some of the hepatocytes were swollen and necrotized by HE staining. A large number of inflammatory cells infiltrated between the portal area and the lobules. The structure of the normal hepatic lobules was obviously disordered. Masson staining showed that the hepatic tissues were proliferated obviously, and the liver tissues were separated by collagen fibers to form pseudolobules. The results showed that the level of alt AST in CBD-VEGF group was higher than that in NS control group 4 weeks after injection. However, there was no significant difference in alt AST levels between the two groups at the 8th week. The liver inflammation degree in the CBD-VEGF group was significantly lighter than that in the NS control group at 4 and 8 weeks after injection with HE staining, which showed that the hepatic fibrosis degree in the CBD-VEGF group was significantly less than that in the NS control group. Immunohistochemistry showed that the number of microvessels in CBD-VEGF group was significantly more than that in NS control group, the activation degree of hepatic stellate cells was lower than that in NS control group, and the proliferation of hepatocytes was higher than that in NS control group. Tunel staining showed that the apoptosis of hepatocytes in CBD-VEGF group was significantly higher than that in NS control group. Conclusion Local injection of CBD-VEGF into liver tissue can significantly attenuate the anti-fibrosis effect of CC14 induced liver fibrosis in mice and promote the regeneration of capillaries in liver tissue. Promoting hepatocyte proliferation, decreasing hepatocyte apoptosis and inhibiting hepatic stellate cell (HSCs) activation are related.
【学位授予单位】：南京大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R575.2

【参考文献】