發(fā)布時(shí)間：2018-02-03 07:03

  本文關(guān)鍵詞： HO-1基因 腸瘺 細(xì)胞移植 骨髓間充質(zhì)干細(xì)胞　出處：《山東大學(xué)》2010年碩士論文　論文類型：學(xué)位論文

【摘要】： 目的：腸瘺是一種嚴(yán)重的術(shù)后并發(fā)癥且死亡率高。主要表現(xiàn)為：瘺口形成與腸內(nèi)容物漏出、感染、營(yíng)養(yǎng)不良、水電解質(zhì)和酸堿平衡紊亂以及多器官功能障礙等。營(yíng)養(yǎng)支持、抗感染、減少腸液分泌、封堵瘺管、維持內(nèi)環(huán)境穩(wěn)定、促進(jìn)瘺管愈合以及選擇性手術(shù)治療等綜合措施,可以提高治愈率。血紅素加氧酶-Ⅰ(HO-1)也稱為熱休克蛋白32,是血紅素降解過(guò)程中的限速酶,它可以降解衰老或破損紅細(xì)胞釋放出的血紅素,生成膽綠素、膽紅素、一氧化碳(CO)和鐵蛋白。大量事實(shí)證明,HO-1在保持細(xì)胞穩(wěn)態(tài)方面能起到極其重要的作用,可以減輕由免疫介導(dǎo)、氧化應(yīng)激、炎性細(xì)胞浸潤(rùn)導(dǎo)致的組織損傷。腺相關(guān)病毒(adeno-associated virus,AAV)是安全有效的基因載體,且有外源基因可以長(zhǎng)期穩(wěn)定表達(dá),分裂期細(xì)胞和靜止期細(xì)胞均能被感染等優(yōu)點(diǎn),還能將基因轉(zhuǎn)人非周期(noncyling)腫瘤細(xì)胞。骨髓間充質(zhì)干細(xì)胞屬于多潛能干細(xì)胞,分化潛能更廣,具有自我更新能力及多向分化能力。能在合適的分化條件下不僅可以分化為成骨細(xì)胞、脂肪細(xì)胞、軟骨細(xì)胞、肌肉細(xì)胞等中胚層細(xì)胞,而且可以分化為外胚層和內(nèi)胚層的細(xì)胞。它成為多種組織修復(fù)的種子細(xì)胞與基因治療的良好載體。在嚴(yán)重的組織缺血和損傷的病例中可觀察到骨髓間充質(zhì)干細(xì)胞遷徙至損傷部位,并分泌生物活性因子,促進(jìn)修復(fù)與再生。骨髓間充質(zhì)干細(xì)胞可以通過(guò)骨髓遷徙或體外擴(kuò)增后直接或全身注射到達(dá)損傷部位,抑制疤痕與凋亡,促進(jìn)血管生成及祖細(xì)胞分裂分化,有利于損傷的愈合。本文探討轉(zhuǎn)染HO-1基因的骨髓間充質(zhì)干細(xì)胞移植對(duì)大鼠腸瘺模型的影響。 方法：骨髓間充質(zhì)干細(xì)胞的分離與培養(yǎng)。骨髓間充質(zhì)干細(xì)胞通過(guò)腺相關(guān)病毒介導(dǎo)轉(zhuǎn)染H0-1基因。以Wistar大鼠建立腸瘺模型,實(shí)驗(yàn)動(dòng)物隨機(jī)分為實(shí)驗(yàn)組(術(shù)后24小時(shí)造瘺口注射1×106個(gè)轉(zhuǎn)染目的基因的骨髓干細(xì)胞),,對(duì)照組(術(shù)后24小時(shí)造瘺口注射生理鹽水100ul)。移植70天后觀察瘺口愈合情況。 結(jié)果：實(shí)驗(yàn)組瘺口愈合明顯快于對(duì)照組。 結(jié)論：轉(zhuǎn)染H0-1基因的骨髓間充質(zhì)干細(xì)胞移植有利于大鼠腸瘺模型瘺口的愈合。
[Abstract]:Objective: intestinal fistula is a serious postoperative complication with high mortality. The main manifestations are fistula formation, leakage of intestinal contents, infection and malnutrition. Water, electrolyte, acid-base balance disorder and multiple organ dysfunction. Nutritional support, anti-infection, reduce intestinal secretion, close fistula, maintain the stability of the internal environment. Heme oxygenase-I (HO-1), also known as heat shock protein 32 (HSP32), is a rate-limiting enzyme in heme degradation. It degrades heme released by aging or damaged red blood cells and produces bilirubin, carbon monoxide (CO) and ferritin. HO-1 can play an extremely important role in maintaining the homeostasis of cells and can alleviate the immune-mediated oxidative stress. Adeno-associated virus (AAVV) is a safe and effective gene vector. Moreover, it has the advantages of long-term stable expression of exogenous genes and infection of both mitotic and stationary cells. Bone marrow mesenchymal stem cells (BMSCs) are multipotent stem cells and have a wider differentiation potential. It can not only differentiate into osteoblasts, adipocytes, chondrocytes, muscle cells and other mesoderm cells, but also can differentiate into osteoblasts, adipocytes, chondrocytes, muscle cells and so on. And it can differentiate into ectodermal and endoderm cells. It is a good carrier of seed cells and gene therapy for multiple tissue repair. Bone marrow mesenchymal stem cells can be observed in severe tissue ischemia and injury cases. Migrate to the site of the injury. Bone marrow mesenchymal stem cells (BMSCs) can be directly or completely injected into the injured site after bone marrow migration or expansion in vitro to inhibit scar and apoptosis. The effects of bone marrow mesenchymal stem cell transplantation (BMSCs) transfected with HO-1 gene on intestinal fistula in rats were studied. Methods: bone marrow mesenchymal stem cells were isolated and cultured. Bone marrow mesenchymal stem cells were transfected with H0-1 gene mediated by adeno-associated virus. Intestinal fistula model was established in Wistar rats. The experimental animals were randomly divided into experimental group (1 脳 106 bone marrow stem cells transfected with target gene) were injected 24 hours after operation. The control group (24 hours after operation) was injected with normal saline 100 ulus. The healing of fistula was observed 70 days after transplantation. Results: the healing of fistula in the experimental group was faster than that in the control group. Conclusion: transplantation of bone marrow mesenchymal stem cells transfected with H0-1 gene is beneficial to the healing of fistula in rat intestinal fistula model.
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R329

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本文編號(hào)：1486759