本文關(guān)鍵詞： 內(nèi)毒素 肝星狀細(xì)胞 髓樣分化因子88 toll樣受體4　出處：《暨南大學(xué)》2014年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：目的 觀察不同濃度內(nèi)毒素對(duì)TLR4和MyD88，及其依賴(lài)MyD88信號(hào)通路上多種細(xì)胞因子表達(dá)的影響，以及對(duì)大鼠肝星狀細(xì)胞（HSCs）增殖與凋亡的影響，探討MyD88在內(nèi)毒素誘導(dǎo)HSCs凋亡信號(hào)轉(zhuǎn)導(dǎo)中的作用，，探索肝纖維化發(fā)病機(jī)制。 方法 體外培養(yǎng)大鼠HSC株HSC-T6，取細(xì)胞濃度與吸光值呈直線關(guān)系且斜率最大處的細(xì)胞濃度作為接種濃度，將內(nèi)毒素按濃度0.1，1.0，10.0EIU/mL（分別為內(nèi)毒素I組、II組和III組）各作用于HSCs，以未加干預(yù)的HSCs為正常對(duì)照組(第IV組)，采用RT-PCR及WesternBlot術(shù)分別檢測(cè)TLR4和MyD88的mRNA與蛋白表達(dá)的變化，以及依賴(lài)MyD88信號(hào)通路的相關(guān)因子TRAF6、TNF-α、NF-кΒ的mRNA與蛋白表達(dá)的變化，并同時(shí)以流式細(xì)胞術(shù)觀察HSC凋亡的變化。 結(jié)果 1.內(nèi)毒素I組、II組和III組中，MyD88、TLR4、NF-κB的mRNA及蛋白的表達(dá)量較對(duì)照組均有所增高（P0.05），其中，內(nèi)毒素II組升高更明顯（P0.01）。 2.內(nèi)毒素I組、II組中，TRAF-6、TNF-α的mRNA及蛋白的表達(dá)量較對(duì)照組有所增高（P0.05），而在內(nèi)毒素III組中，其mRNA及蛋白的表達(dá)量無(wú)明顯影響（P＞0.05）。 3.內(nèi)毒素I組、II組中HSC的平均吸光度顯著高于對(duì)照組(P0.05)。而毒素III組中HSC的平均吸光度無(wú)明顯增高(P＞0.05)，提示一定濃度的內(nèi)毒素能夠促進(jìn)肝星狀細(xì)胞的增殖。 4.內(nèi)毒素I組、II組中HSC凋亡減少（P0.05），其中內(nèi)毒素II組中HSC凋亡減少更明顯，（P0.01），而內(nèi)毒素III組中HSC凋亡無(wú)明顯影響（P0.05）。 結(jié)論 一定濃度的內(nèi)毒素可以促進(jìn)TLR4、MyD88及其信號(hào)通路相關(guān)因子TRAF6、TNF-α、NF-кΒ的表達(dá)，促進(jìn)肝星狀細(xì)胞的增殖，抑制其凋亡，內(nèi)毒素可能由此機(jī)制促進(jìn)肝纖維化發(fā)生發(fā)展。
[Abstract]:Purpose. To investigate the effects of different concentrations of endotoxin on the expression of cytokines in TLR4, MyD88and its dependent MyD88 signaling pathway, and on the proliferation and apoptosis of rat hepatic stellate cells (HSCs), and to explore the role of MyD88 in the signal transduction of HSCs apoptosis induced by endotoxin. To explore the pathogenesis of liver fibrosis. Method. Rat HSC strain HSC-T6 was cultured in vitro. The cell concentration which had a linear relationship with the absorptivity and the highest slope was taken as the inoculation concentration. Endotoxin was treated with 0.1 ~ 1.0 ~ 1.0 ~ 10.0 EIU-mL (endotoxin I group II and III group) respectively, and HSCs without intervention was used as normal control group (group IV). The changes of mRNA and protein expression of TLR4 and MyD88 were detected by RT-PCR and WesternBlot, respectively. The changes of mRNA and protein expression of TRAF6TNF- 偽 TNF- 偽 尾 were also observed by flow cytometry. The apoptosis of HSC was also observed by flow cytometry. Results. 1. The expression of mRNA and protein of MyD88TLR4NF- 魏 B in group I and group III were higher than those in control group (P 0.05), especially in group II (P 0.01). 2. The expression of mRNA and protein of TRAF-6 TNF- 偽 in lipopolysaccharide I group was higher than that in control group (P 0.05), but the expression of mRNA and protein in endotoxin III group had no significant effect (P > 0.05). 3. The average absorbance of HSC in group I was significantly higher than that in control group (P 0.05), but the average absorbance of HSC in III group was not significantly increased (P > 0.05), suggesting that endotoxin at a certain concentration could promote the proliferation of hepatic stellate cells. 4. The apoptosis of HSC in group I was decreased by P0.05, and the apoptosis of HSC was decreased more significantly in group II, but the apoptosis of HSC in group III was not significantly affected by P0.05. Conclusion. A certain concentration of endotoxin can promote the expression of TLR4- MyD88 and its signal transduction related factor TRAF6- TNF- 偽 -NF- 尾, promote the proliferation of hepatic stellate cells and inhibit its apoptosis. Endotoxin may promote the occurrence and development of hepatic fibrosis.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R575.2

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