本文選題：Sept4 + mi��； 參考：《南通大學》2014年碩士論文

【摘要】：目的探索Sept4對日本血吸蟲病小鼠慢性肝纖維化的作用;同時觀察miR-454在活化的肝星狀細胞HSCs和日本血吸蟲誘導的肝纖維化組織中的表達變化及其對HSCs的影響和機制。方法1.ICR小鼠30只,適應性喂養(yǎng)1w后,隨機分為3組,即PBS組(血吸蟲感染+PBS處理組),Ad-GFP組(血吸蟲感染+Ad-GFP處理組),Ad-Sept4組(血吸蟲感染+Ad-Sept4處理組)。各組小鼠感染血吸蟲,12w后經(jīng)吡喹酮殺蟲治療3d(250mg/kg/24h),然后經(jīng)尾靜脈分別給各小鼠注射Ad-Sept4、Ad-GFP或PBS。于注射2w后(即感染約15w后)處死小鼠,取肝組織進行HE染色和天狼星紅染色,觀察肝纖維化組織的變化情況;用Western Blot和qRT-PCR技術(shù)檢測肝纖維化相關(guān)基因的表達變化情況;用TUNEL觀察凋亡情況。2.ICR小鼠16只,適應性喂養(yǎng)1w后,隨機分為2組,即正常組(未感染)和感染組(血吸蟲感染),感染組在感染日本血吸蟲8w后處死小鼠,取肝臟組織以備用。應用qRT-PCR技術(shù)檢測miR-454在肝纖維化組織中及活化的HSCs中的表達變化;用MTT、流式細胞術(shù)檢測miR-454對HSCs細胞增殖和細胞周期的影響;應用Western Blot檢測mi R-454對HSCs活化的影響。結(jié)果1.與Ad-GFP組相比,Ad-Sept4組的肝臟纖維化程度減輕;TUNEL陽性細胞數(shù)量增加;Sept4及cleaved-caspase-3表達上升;而α-SMA、pro-collα1(I)、TGF-β1和IL-6等表達下降。2.在肝纖維化組織中及活化的HSCs中miR-454的表達下降,α-SMA和Smad4表達上升;過表達mi R-454可以降低HSCs中的α-SMA和Smad4表達,但不能影響HSCs的增殖和細胞周期。3.在活化的HSCs中,miR-454可以抑制Smad4 3’UTR端的熒光素酶活性。結(jié)論1.Ad-Sept4對血吸蟲病小鼠慢性肝纖維化具有抑制作用。2.miR-454在血吸蟲病肝纖維化組織及活化HSCs中表達降低。3.過表達miR-454可以通過靶向Smad4 3’UTR區(qū)發(fā)揮抑制HSCs活化的作用。
[Abstract]:Objective to investigate the effect of Sept4 on chronic hepatic fibrosis in mice with schistosomiasis japonicum, and to observe the expression of miR-454 in activated hepatic stellate cells (HSCs) and Schistosoma japonicum induced hepatic fibrosis (HSCs). Methods Thirty 1.ICR mice were randomly divided into three groups after one week of adaptive feeding: PBS group (PBS treatment group) (Ad-GFP treatment group) and Ad-Sept4 group (Ad-Sept4 treatment group). The mice in each group were infected with Schistosoma japonicum for 12 weeks and treated with praziquantel for 3 days, 250 mg / kg / 24 h / h, and then each mouse was injected with Ad-Sept4Ad-GFP or PBSvia caudal vein respectively. The mice were killed 2 weeks after injection (that is, 15 weeks after infection), the liver tissues were stained with HE and Sirius red to observe the changes of liver fibrosis tissues, and the expression of genes related to liver fibrosis were detected by Western Blot and qRT-PCR techniques. After 1 week of adaptive feeding, 16 ICR mice were randomly divided into two groups: normal group (uninfected) and infected group (Schistosoma japonicum infection). The infected mice were killed after 8 weeks of infection with Schistosoma japonicum, and liver tissues were taken for reserve. QRT-PCR technique was used to detect the expression of miR-454 in liver fibrosis tissues and activated HSCs, MTT and flow cytometry were used to detect the effect of miR-454 on the proliferation and cell cycle of HSCs cells, and Western Blot was used to detect the effect of miR-454 on HSCs activation. Result 1. Compared with Ad-GFP group, the degree of hepatic fibrosis in Ad-Sept4 group reduced the number of Tunel positive cells increased, while the expression of 偽 -SMA-pro-coll 偽 1 and TGF- 尾 1 and IL-6 decreased .2.The expression of TGF- 尾 1 and TGF- 尾 1 were decreased in Ad-Sept4 group, and the expression of TGF- 尾 1 and IL-6 were decreased. The expression of 偽 -SMA and Smad4 increased in hepatic fibrosis tissues and activated HSCs, and the overexpression of miR-454 could decrease the expression of 偽 -SMA and Smad4 in HSCs, but it could not affect the proliferation and cell cycle of HSCs. In activated HSCs, miR-454 inhibited luciferase activity at the end of Smad4 3'UTR. Conclusion 1.Ad-Sept4 can inhibit chronic hepatic fibrosis in mice with schistosomiasis. 2. The expression of miR-454 in schistosomiasis liver fibrosis tissue and activated HSCs is decreased. Overexpression of miR-454 can inhibit the activation of HSCs by targeting the region of Smad4 3'UTR.
【學位授予單位】：南通大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R532.21

【參考文獻】

相關(guān)期刊論文 前1條

1 張志安,石淑仙,黃完成;活血化瘀類中藥對實驗性血吸蟲肝纖維化兔肝組織金屬蛋白酶-1及其抑制因子-1mRNA表達的影響[J];中西醫(yī)結(jié)合肝病雜志;2001年06期

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