目的通過建立大鼠肝纖維化及IL-10干預(yù)模型,研究肝纖維化大鼠肝組織中IL-10的表達(dá)和變化,探討外源性IL-10對(duì)肝細(xì)胞的作用;通過外源性IL-10對(duì)原代肝細(xì)胞進(jìn)行培養(yǎng)干預(yù),了解IL-10對(duì)肝細(xì)胞增殖、凋亡的影響,并分析其相關(guān)生長(zhǎng)因子的表達(dá)變化。方法動(dòng)物實(shí)驗(yàn)部分:建立大鼠肝纖維化和IL-10干預(yù)模型,抽提肝臟總RNA,Real-Time PCR法檢測(cè)模型組與正常組IL-10表達(dá)的差異,原位Tunnel法檢測(cè)IL-10對(duì)肝細(xì)胞凋亡的影響;免疫組織化學(xué)法、RT-PCR檢測(cè)肝組織Bax/Bcl-2蛋白及mRNA的表達(dá)。細(xì)胞實(shí)驗(yàn)部分:通過膠原蛋白酶原位灌流法分離大鼠原代肝細(xì)胞, RT-PCR法檢測(cè)肝內(nèi)不同細(xì)胞標(biāo)志基因,如:ALB、CK19、AFP、CYP8B1、CD68、CD31等基因的表達(dá)情況,并進(jìn)行細(xì)胞純度鑒定、凍存研究、復(fù)蘇培養(yǎng)、細(xì)胞活性鑒定等;RT-PCR、Western-blot檢測(cè)原代肝細(xì)胞IL-10/IL-10R1 mRNA和蛋白的表達(dá)情況;原代肝細(xì)胞分三組培養(yǎng):正常組（N組,普通培養(yǎng)基）、對(duì)照組（C組,含胰島素培養(yǎng)基）和IL-10干預(yù)組（I組,含IL-10、胰島素的培養(yǎng)基）,... 

【文章來源】：福建醫(yī)科大學(xué)福建省

【文章頁(yè)數(shù)】：63 頁(yè)

【學(xué)位級(jí)別】：博士

【部分圖文】：

組IL-10mRNA、β-actin電泳圖

組IL-10mRNA、β-actin電泳圖

（0.72±0.25）%，M 組見多量陽性細(xì)胞散在（18.87±3.58）%，S 組較 N 組陽細(xì)胞數(shù)增多，兩者差異顯著（P<0.01）；T 組陽性率（4.77±2.15）%，較 R 組（±1.85）%，陽性細(xì)胞數(shù)顯著減少（19.23%，P<0.01），（圖 4）。

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