本文關(guān)鍵詞：肝細(xì)胞生長(zhǎng)因子轉(zhuǎn)染的MSCs防治小鼠腸道移植物抗宿主病及其機(jī)理研究　出處：《中國(guó)人民解放軍軍事醫(yī)學(xué)科學(xué)院》2006年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 造血干細(xì)胞移植 移植物抗宿主病 間充質(zhì)干細(xì)胞 肝細(xì)胞生長(zhǎng)因子

【摘要】：異基因造血干細(xì)胞移植是治療惡性血液病最有效的方法之一，但是，由于供受者之間主要組織相容性復(fù)合體的不同，移植物抗宿主病(GVHD)的發(fā)生難以完全避免。GVHD可以影響多個(gè)器官，主要是皮膚、肝臟和腸道。重度腸道GVHD會(huì)加重全身GVHD，影響移植的療效，增加病死率。腸道粘膜損傷能夠通過加重GVHD時(shí)的炎性細(xì)胞因子風(fēng)暴，因此，粘膜損傷修復(fù)和免疫調(diào)節(jié)是腸道GVHD治療的關(guān)鍵。 肝細(xì)胞生長(zhǎng)因子(HGF)對(duì)許多組織都具有修復(fù)作用，間充質(zhì)干細(xì)胞(MSCs)不僅具有損傷修復(fù)作用，還具有免疫調(diào)節(jié)、促進(jìn)造血恢復(fù)的作用，本實(shí)驗(yàn)建立小鼠異基因造血干細(xì)胞移植腸道GVHD模型；應(yīng)用高表達(dá)HGF的MSCs(MSC-HGF)防治小鼠腸道GVHD，比較其與藥物以及單獨(dú)應(yīng)用MSC防治GVHD之間的差異。 材料和方法Balb／c小鼠經(jīng)5.5Gy照射后輸注C57BL／6小鼠骨髓細(xì)胞及脾細(xì)胞建立小鼠腸道GVHD模型；分離培養(yǎng)、鑒定C57BL／6小鼠骨髓MSCs，應(yīng)用脂質(zhì)體法將質(zhì)粒pcDNA3.1-HGF瞬時(shí)轉(zhuǎn)染入MSCs，檢測(cè)轉(zhuǎn)染細(xì)胞HGF表達(dá)情況；實(shí)驗(yàn)組根據(jù)細(xì)胞輸注時(shí)間(0d，+6d，+10d)和輸注形式(MSC或MSC-HGF)的不同分為6組：MSCs-0d，MSC-HGF-0d，MSCs-6d，MSC-HGF-6d，MSCs-10d，MSC-HGF-10d；輸注MSCs細(xì)胞數(shù)量為1×10~6／只：空白對(duì)照組未予GVHD預(yù)防與治療，藥物對(duì)照組采用環(huán)胞菌素(CSA，5 mg／kg／d，灌胃)和驍悉(MMF，40 mg／kg／d，灌胃)聯(lián)合防治GVHD。比較移植后腸道GVHD的發(fā)生率和致死率，腸道病理損傷情況以及小鼠存活情況。檢測(cè)細(xì)胞輸注后受體小鼠T細(xì)胞亞群變化，CD4+CD25+T細(xì)胞比例變化，，炎癥相關(guān)細(xì)胞因子表達(dá)情況，以及腸道凋亡細(xì)胞變化。
[Abstract]:Allogeneic hematopoietic stem cell transplantation (HSCT) is one of the most effective methods for the treatment of malignant hematological diseases, but due to the difference of major histocompatibility complex among donors. Graft-versus-host disease (GVHD) is difficult to avoid completely. GVHD can affect many organs, mainly skin, liver and intestine. Severe intestinal GVHD can aggravate systemic GVHD. Intestinal mucosal injury can be induced by inflammatory cytokines storm in GVHD. Therefore, mucosal damage repair and immunomodulation are the key factors in the treatment of intestinal GVHD. Hepatocyte growth factor (HGF) has repair effect on many tissues, mesenchymal stem cells (MSCs) have not only damage repair, but also immune regulation, promote hematopoietic recovery. The GVHD model of allogeneic hematopoietic stem cell transplantation was established in this experiment. High expression of HGF MSC-HGF-HGFH was used to prevent and treat GVHD in mouse intestine, and the difference was compared with drug and MSC alone in the prevention and treatment of GVHD. Materials and methods Balb/c mice were irradiated with 5.5 Gy and then injected with C57BL / 6 mice bone marrow cells and spleen cells to establish the intestinal GVHD model of mice. C57BL / 6 mouse bone marrow MSCs were isolated and identified. The plasmid pcDNA3.1-HGF was transiently transfected into MSCs by liposome method. The expression of HGF in transfected cells was detected. The experimental group was divided into 6 groups according to the time of cell infusion (0 d, 6 d, 10 d) and the form of infusion (MSC or MSC-HGF) into 6 groups: MSC-HGF-0d. MSCs-6dT MSC-HGF-6dT; MSCs-10d; MSC-HGF-10d; The number of MSCs cells was 1 脳 10 ~ (-6) / mouse: the blank control group was not treated with GVHD, while the drug control group was treated with cyclosporine for 5 mg/kg/d. The incidence and mortality of intestinal GVHD after transplantation were compared with 40 mg / kg / d of mycophenolate mofetil and 40 mg / kg / d of mycophenolate mofetil. The changes of T cell subsets in recipient mice and the percentage of CD4 CD25 T cells and the expression of inflammatory cytokines were detected. And the changes of intestinal apoptotic cells.
【學(xué)位授予單位】：中國(guó)人民解放軍軍事醫(yī)學(xué)科學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2006
【分類號(hào)】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 劉麗輝,孫琪云,胡鍇勛,黃雅靜,范傳波,孫昭,姚波,郭梅,趙春華,艾輝勝;獼猴單倍體相合非清髓造血干細(xì)胞聯(lián)合間充質(zhì)干細(xì)胞移植的實(shí)驗(yàn)研究[J];中華血液學(xué)雜志;2005年07期

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