本文選題：肝大部切除鼠血清 + 骨髓干細(xì)胞　； 參考：《復(fù)旦大學(xué)》2005年博士論文

【摘要】：近年來的研究提示,成體骨髓干細(xì)胞(adult bone marrow stem cell,ABMSC)能轉(zhuǎn)分化為肝細(xì)胞。這一現(xiàn)象為臨床上應(yīng)用ABMSC移植治療肝損傷和急慢性肝功能衰竭展示了一個可喜的前景。但是ABMSC向肝細(xì)胞分化的影響因素及分化的機制尚不十分清楚。我們課題組先前有關(guān)肝再生的研究資料和國內(nèi)外有關(guān)研究進展提示,雖然肝細(xì)胞生長因子(hepatocyte growth factor,HGF)是啟動肝細(xì)胞DNA合成的主要因子,但肝細(xì)咆的生長、分化和增殖要受到機體整體狀態(tài)、以及肝外和肝內(nèi)多種因子和某些激素等的調(diào)控,而肝大部切除術(shù)后的血清則可能包含全部刺激因子和營養(yǎng)成分。因此本課題應(yīng)用肝大部切除鼠血清和/或HGF,進行ABMSC向肝細(xì)胞分化的體內(nèi)外實驗研究。 研究目的 1.應(yīng)用肝大部切除鼠血清和/或HGF在體外誘導(dǎo)同系A(chǔ)BMSC向肝細(xì)胞分化,以探討ABMSC向肝細(xì)胞前體分化和擴增的體外培養(yǎng)方法和條件。 2.誘導(dǎo)分化至一定階段的大鼠ABMSC經(jīng)不同途徑(靜脈注射、肝內(nèi)注射和脾內(nèi)注射)回輸?shù)酵荡笫蟾螕p傷動物模型體內(nèi),觀察其在體內(nèi)向肝細(xì)胞分化以及參與肝組織重建的情況。 3.運用基因芯片技術(shù)對肝大部切除鼠血清誘導(dǎo)ABMSC向肝細(xì)胞分化的基因表達譜進行篩選,分析發(fā)生表達變化的基因,以期了解與ABMSC向肝細(xì)胞分化相關(guān)的特異性基因及它們之間的相互關(guān)系,從基因水平揭示了ABMSC向肝細(xì)胞分化中的分子機制。 研究方法 通過肝大部切除手術(shù),制備肝再生動物模型,在術(shù)后24小時收集肝大部切除鼠血清。分離骨髓細(xì)胞體外培養(yǎng),應(yīng)用肝大部切除鼠血清誘導(dǎo)分化,利用免疫組化,RT-PCR,Western Blot等技術(shù)鑒定分化細(xì)胞。將已分化至一定階段的肝前體細(xì)胞用CM-DiI進行標(biāo)記后,回輸?shù)礁未蟛壳谐母卧偕鷦游锬Ｐ腕w內(nèi),通過免疫組化和激光共聚焦顯微鏡等方法觀察其在體內(nèi)重建肝組織的情況。利用基因
[Abstract]:Recent studies suggest that adult bone marrow stem cells can be transformed into hepatocytes. This phenomenon presents a promising prospect for the clinical application of ABMSC transplantation in the treatment of liver injury and acute and chronic liver failure. However, the factors influencing the differentiation of ABMSC into hepatocytes and the mechanism of differentiation are not well understood. Our previous research data on liver regeneration and research progress at home and abroad suggest that although hepatocyte growth factor (HGF) is the main factor to initiate the synthesis of hepatocyte DNA, the growth of liver cells can be observed. Differentiation and proliferation are regulated by the whole state of the body, various factors and some hormones in the extrahepatic and intrahepatic liver, while the serum after subtotal hepatectomy may contain all the stimulating factors and nutrients. Therefore, the in vitro and in vivo study of the differentiation of ABMSC into hepatocytes was carried out by using rat serum and / or HGF after subtotal hepatectomy. Hepatectomy rat serum and / or HGF were used to induce the differentiation of homologous ABMSC into hepatocytes in vitro, in order to explore the culture methods and conditions for the differentiation and expansion of ABMSC into hepatocyte precursors in vitro. 2. After induction and differentiation to a certain stage, ABMSC was infused into the same rat model of liver injury by different ways (intrahepatic injection, intrahepatic injection and intrasplenic injection). To observe its differentiation into hepatocytes in vivo and participate in the reconstruction of liver tissue. 3. 3. The gene expression profiles of hepatocyte differentiation induced by serum from subtotal hepatectomy rats were screened by gene chip technique, and the genes that changed in expression were analyzed. In order to understand the specific genes related to the differentiation of ABMSC into hepatocytes and their relationship, the molecular mechanism of differentiation of ABMSC into hepatocytes was revealed at the gene level. Animal models of liver regeneration were established and serum samples were collected at 24 hours after hepatectomy. Bone marrow cells were isolated and cultured in vitro. Differentiation was induced by rat serum after subtotal hepatectomy. The differentiated cells were identified by immunohistochemistry with RT-PCR- Western blot. The liver precursor cells which had differentiated to a certain stage were labeled with CM-DiI and then transferred back to the model of liver regeneration after subtotal hepatectomy. The reconstitution of liver tissue in vivo was observed by immunohistochemistry and confocal laser microscopy. Utilization gene
【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2005
【分類號】：R329

【參考文獻】

相關(guān)期刊論文 前6條

1 陳平;肝部分切除術(shù)后肝細(xì)胞再生機制的探討[J];第三軍醫(yī)大學(xué)學(xué)報;2003年12期

2 陳偉,張新華,鐘翠平;肝再生的分子生物學(xué)研究進展[J];解剖科學(xué)進展;2003年03期

3 張雪梅,劉秉文;載脂蛋白E研究進展[J];中國動脈硬化雜志;2001年02期

4 劉希琴，，成令忠;肝大部切除后肝再生調(diào)節(jié)的研究[J];生理科學(xué)進展;1995年04期

5 劉艷環(huán),蘇長來;ApoE基因多態(tài)性和運動與脂質(zhì)代謝及冠心病的研究進展[J];天津體育學(xué)院學(xué)報;2003年03期

6 王宇明,陳耀凱;肝細(xì)胞移植的進展與挑戰(zhàn)[J];中華肝臟病雜志;2003年06期

本文編號：1987465