本文關(guān)鍵詞： 骨髓間充質(zhì)干細(xì)胞 缺血再灌注損傷 Survinvin 存活能力　出處：《重慶醫(yī)科大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：探討轉(zhuǎn)染survinvin基因后的骨髓間充質(zhì)干細(xì)胞（MSCs)在腎臟缺血微環(huán)境中的存活能力以及對(duì)小鼠腎臟缺血再灌注損傷的修復(fù)作用能力和機(jī)制。 方法：復(fù)蘇、擴(kuò)增培養(yǎng)已完成空病毒、Survinvin基因轉(zhuǎn)染、鑒定、EGFP標(biāo)記工作的MSCs，，擴(kuò)增培養(yǎng),觀察綠色熒光表達(dá)情況備用，48只SPF級(jí)健康雄性C57BL/6J分為正常對(duì)照組12只、IR組（小鼠雙側(cè)腎動(dòng)脈夾閉40min后恢復(fù)血供）12只、空病毒轉(zhuǎn)染移植組(MSCs組，小鼠雙側(cè)腎動(dòng)脈夾閉40min后恢復(fù)血供24后經(jīng)尾靜脈注射200微升1×106個(gè)細(xì)胞)12只、survinvin基因轉(zhuǎn)染移植組(SVV/MSCs組，小鼠雙側(cè)腎動(dòng)脈夾閉40min后恢復(fù)血供24小時(shí)后經(jīng)尾靜脈注射200微升1×106個(gè)survinvin基因轉(zhuǎn)染細(xì)胞)24只，選取細(xì)胞移植后1d、3d、7d、14d不同時(shí)間點(diǎn)采集血清行肌酐尿素氮檢測(cè)，切取小鼠腎臟行石蠟切片觀察MSCs存活數(shù)目并定量分析，HE染色觀察腎組織病理變化及腎小管損害程度評(píng)分，ELISA檢測(cè)腎臟勻漿中細(xì)胞因子HGF、bFGF、IL-10的表達(dá)情況。 結(jié)果：復(fù)蘇、擴(kuò)增培養(yǎng)出的MSCs增值活力旺盛，熒光狀態(tài)良好；移植細(xì)胞1天后MSCs組與SVV/MSCs組肌酐尿素氮水平明顯低于IR對(duì)照組（p0.01或p0.05）。腎臟HE染色病理?yè)p害評(píng)分,干細(xì)胞移植組損傷較輕，SVV/MSCs明顯低于其他兩組（p0.01或p0.05）。第3天、14天存活于腎組織的移植細(xì)胞數(shù)目SVV/MSCs組遠(yuǎn)高于MSCs組，表達(dá)EGFP的MSCs主要分布于腎小球周圍、小血管內(nèi)壁、腎小管與腎小管之間的間質(zhì),而腎小管內(nèi)壁幾乎見不到表達(dá)EGFP的MSCs。在腎臟缺血損傷后保護(hù)性因子HGF、bFGF、IL-10水平升高SVV/MSCs也比IR組、MSCs組明顯的多（P0.01或p0.05)，并且在第14天時(shí)與正常對(duì)照組已無(wú)統(tǒng)計(jì)學(xué)差異。 結(jié)論:轉(zhuǎn)染Survinivin基因可以增加MSCs在缺血腎臟中的生存能力，MSCs主要通過其強(qiáng)大的旁分泌作用進(jìn)一步促進(jìn)腎臟損傷的修復(fù)。
[Abstract]:Aim: to investigate the viability of bone marrow mesenchymal stem cells (MSCs) transfected with survinvin gene in renal ischemic microenvironment and the repair ability and mechanism of renal ischemia-reperfusion injury in mice. Methods: resuscitation, amplification and culture of empty virus Survinvin gene transfection, identification of EGFP labeled MSCs, amplification of culture, To observe the expression of green fluorescence, 48 healthy male C57BL / 6J of SPF grade were divided into 12 normal control group (12 normal control group) (12 normal control group) (12 mice recovered blood supply after bilateral renal artery occlusion for 40 minutes, and 12 mice were treated with empty virus transfection and transplantation group). After bilateral renal artery occlusion for 40 minutes, the blood supply was restored 24 minutes later, and then injected with 200 microliters of 1 脳 106 cells through tail vein. 12 mice were transfected with survinvin gene into SVV / MSCs group. After bilateral renal artery occlusion for 40 minutes, the blood supply was restored 24 hours later, and then injected into tail vein of 24 mice with 1 脳 106 survinvin gene transfection. Serum creatinine urea nitrogen was measured at different time points of 1 day, 3 days, 7 days and 14 days after transplantation. The survival number of MSCs was observed by paraffin section and the pathological changes of renal tissue and the expression of cytokine HGFbFGF- IL-10 in renal homogenate were detected by Elisa. Results: after resuscitation, the proliferative activity and fluorescence state of MSCs were exuberant, the level of creatinine urea nitrogen in MSCs group and SVV/MSCs group was significantly lower than that in IR control group (p0.01 or p0.05), and the pathological damage score of renal HE staining was significantly lower than that in MSCs group and SVV/MSCs group. The number of transplanted cells survived in renal tissue in SVV/MSCs group was much higher than that in MSCs group on the 3rd day after transplantation. The MSCs expressing EGFP was mainly distributed around glomeruli and in the inner wall of small blood vessels, and the number of transplanted cells was significantly lower in the stem cell transplantation group than in the other two groups (P < 0.01 or P 0.05), and the number of the transplanted cells in the renal tissue was significantly higher in the SVV/MSCs group than in the MSCs group. The interstitium between the tubules and the renal tubules, The level of HGFbFGFGF-IL-10 was significantly higher in the renal tubular wall than that in the IR group, and there was no significant difference between them on the 14th day and the control group. Conclusion: transfection of Survinivin gene can increase the viability of MSCs in ischemic kidney and promote the repair of renal injury mainly through its strong paracrine effect.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R363

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 季衛(wèi)鋒;丁偉航;馬鎮(zhèn)川;厲駒;童培建;;三通道髓芯鉆孔減壓加DBM、自體骨髓干細(xì)胞治療早期股骨頭壞死[J];中國(guó)骨傷;2008年10期

本文編號(hào)：1532730