本文選題：血管內(nèi)皮細胞生長因子　切入點：膠原結(jié)合區(qū)　出處：《南京大學》2014年碩士論文　論文類型：學位論文

【摘要】：目的：探討膠原結(jié)合血管內(nèi)皮細胞生長因子(Collagen-binding-domain vascular endothelial growth factor, CBD-VEGF)抗四氯化碳(CCh)誘導的小鼠肝纖維化作用及其可能機制。方法：BALB/c小鼠腹腔注射CCl4油劑(CCl4：橄欖油=1：5,5μl/g),每周2次,連續(xù)12周建立小鼠肝纖維化模型。將肝纖維化小鼠隨機分為三組：正常對照組、生理鹽水對照組和CBD-VEGF組。CBD-VEGF組肝臟局部注射50山CBD-VEGF (10μl),生理鹽水對照組肝臟局部注射50/μl生理鹽水。注射后4周及8周各組隨機處死小鼠,采集外周血,分離血清,檢測肝功能。同時,取出肝組織,進行HE和Masson染色觀察肝組織形態(tài)和纖維化程度,免疫組織化學方法檢測肝組織中CD31、α-平滑肌激動蛋白(a-SMA)及Ki-67的表達,TUNEL法檢測肝細胞凋亡。結(jié)果CCh腹腔注射12周后,小鼠肝組織HE染色可見部分肝細胞腫脹、壞死,匯管區(qū)和小葉間有大量炎癥細胞浸潤,正常肝小葉結(jié)構(gòu)明顯紊亂。Masson染色見肝組織內(nèi)纖維增生明顯,肝組織被膠原纖維所分隔,形成假小葉,表明小鼠肝纖維化模型建立成功。CBD-VEGF注射后,肝功能檢測結(jié)果顯示,注射后4周,CBD-VEGF組ALT、AST水平高于NS對照組,而8周時兩組ALT、AST水平無統(tǒng)計學差異。肝組織HE染色示,CBD-VEGF注射后4周和8周小鼠肝臟炎癥程度明顯輕于NS對照組。Masson染色表明CBD-VEGF組肝纖維化程度較NS對照組明顯減輕。免疫組織化學表明CBD-VEGF組微血管數(shù)量明顯多于NS對照組(p0.01),肝星狀細胞活化程度低于NS對照組(p0.01),肝細胞增殖高于NS對照組(p0.01)。TUNEL染色表明CBD-VEGF組肝細胞凋亡明顯較NS對照組輕(p0.05)。結(jié)論肝臟組織局部注射CBD-VEGF能夠顯著減輕CC14誘導的小鼠肝纖維化。CBD-VEGF的抗纖維化作用可能與其促進肝組織內(nèi)毛細血管再生,促進肝細胞增殖,減少肝細胞凋亡,抑制肝星狀細胞(hepatic stellate cells, HSCs)活化有關。
[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, 偽 -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.
【學位授予單位】：南京大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R575.2

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本文編號：1561291