本文選題：HLA-G + 腎移植模型��； 參考：《安徽醫(yī)科大學(xué)》2017年碩士論文

【摘要】：【背景】腎移植后排斥反應(yīng)是影響移植受者存活的重要因素,而長期應(yīng)用免疫抑制劑有其不可避免的缺點,因此使移植腎產(chǎn)生免疫耐受是移植界的重要課題。人類白細(xì)胞抗原G(human leukocyte antigen-G,HLA-G)作為一種免疫耐受分子,對母胎耐受及正常妊娠有著極其重要的作用,而其在器官移植中的作用也逐漸被發(fā)現(xiàn),移植術(shù)后患者體內(nèi)HLA-G的表達(dá)已被證實可以降低排斥反應(yīng)的發(fā)生率。通過向大鼠移植腎轉(zhuǎn)染HLA-G基因使其在移植腎內(nèi)高表達(dá),有望成為誘導(dǎo)移植腎免疫耐受的新途徑�！灸康摹勘緦嶒灁M向大鼠移植腎中轉(zhuǎn)染HLA-G基因并將轉(zhuǎn)染過基因的移植腎移植到受體大鼠上,通過檢測大鼠移植腎的病理變化以及HLA-G基因的表達(dá)情況,反應(yīng)出移植腎通過轉(zhuǎn)染HLA-G基因?qū)σ浦残g(shù)后移植腎免疫排斥反應(yīng)的影響,為HLA-G基因誘導(dǎo)移植腎免疫耐受的基礎(chǔ)實驗性研究提供有力的研究工具。【方法】分別建立SD大鼠移植實驗組和對照組,實驗組為轉(zhuǎn)染過含有HLA-G基因的慢病毒,陰性對照組為轉(zhuǎn)染過不含有HLA-G基因的慢病毒,空白對照組為灌注液處理的移植腎,然后分別將三組移植腎通過建立動靜脈袖套的方法將移植腎移植到受體腎動靜脈上,同時將移植腎膀胱瓣膜縫合到受體膀胱壁上完成大鼠腎移植模型的建立。建模成功后分別于造模后一周開始,每周三組隨機(jī)選取三只大鼠,麻醉后沿腹正中切開觀察移植腎變化,處死并取出移植腎,部分放-80℃保存,其余甲醛固定。然后分別對三組移植腎組織進(jìn)行HE染色分析移植腎病理變化,免疫組化以及Western blot分析HLA-G基因的表達(dá)情況,分析出轉(zhuǎn)染HLA-G基因?qū)Υ笫笠浦材I的免疫排斥反應(yīng)的影響。【結(jié)果】共建立大鼠腎移植模型30只,死亡3只,成功率90%,其中2只死于出血,一只死于尿瘺。通過對三組移植腎的病理分析發(fā)現(xiàn),對照組排斥反應(yīng)明顯,而實驗組排斥反應(yīng)明顯低于對照組,同時免疫組化及Western blot分析HLA-G基因的表達(dá)情況發(fā)現(xiàn),對照組均不表達(dá)HLA-G基因,而實驗組均表達(dá)HLA-G基因,而且隨著實踐的延長,表達(dá)量增加�！窘Y(jié)論】本實驗通過建立腎動靜脈袖套方法建立大鼠腎移植模型,有效的減少手術(shù)時間,簡化手術(shù)復(fù)雜程度,為腎移植模型的普通化提供新的思路。同時,實驗還發(fā)現(xiàn)轉(zhuǎn)染HLA-G基因可以有效減少移植后急性免疫排斥反應(yīng)的發(fā)生,延緩慢性免疫排斥反應(yīng),為轉(zhuǎn)染HLA-G基因誘導(dǎo)移植腎免疫耐受的實驗性研究奠定了基礎(chǔ)。
[Abstract]:[background] rejection after renal transplantation is an important factor affecting the survival of recipients, and the long-term application of immunosuppressants has its inevitable shortcomings. As an immunological tolerance molecule, human leukocyte antigen-GG plays an extremely important role in maternal and fetal tolerance and normal pregnancy, and its role in organ transplantation has been gradually discovered. The expression of HLA-G has been shown to reduce the incidence of rejection after transplantation. The high expression of HLA-G gene was induced by transfection of HLA-G gene into the transplanted kidney of rats. [objective] to transfect HLA-G gene into the transplanted kidney of rats and transplant the transfected kidney to the recipient rats. By detecting the pathological changes and the expression of HLA-G gene in the transplanted kidney of rats, the effect of HLA-G gene transfection on the immune rejection of the transplanted kidney was revealed. [methods] A SD rat experimental group and a control group were established respectively. The experimental group was transfected with lentivirus containing HLA-G gene. The negative control group was infected with lentivirus which did not contain HLA-G gene, while the blank control group was treated with perfusion solution. The three groups of transplanted kidneys were transplanted to the recipient renal arteriovenous by establishing an arteriovenous cuff. At the same time, the graft bladder valve was sutured to the recipient bladder wall to complete the establishment of rat renal transplantation model. Three rats were randomly selected each week after modeling. After anesthesia, the allografts were observed by incision along the middle of the abdomen, then killed and removed, some of them were preserved at -80 鈩，

本文編號：1994829