【摘要】：目的 研究人臍血間充質(zhì)干細(xì)胞（human umbilical cord blood mesenchymal stemcells, hUCB-MSCs）移植調(diào)節(jié)兔局灶性腦缺血再灌注損傷早期炎性反應(yīng)和神經(jīng)保護(hù)的機(jī)制。 方法 采集臍帶血80ml-100ml，體外分離hUCB-MSCs并傳代培養(yǎng)，取第6代培養(yǎng)的細(xì)胞做治療移植用。健康新西蘭大白兔64只，隨機(jī)分為假手術(shù)組、缺血再灌注組，MSC組，生理鹽水組，每組16只。采用線栓法建立兔大腦中動(dòng)脈缺血再灌注模型（middle cerebral artery occlusion, MCAO），MSC組通過(guò)右腿的股靜脈移植內(nèi)含3mL5×106個(gè)hUCB-MSCs懸液，生理鹽水組則采用同樣的方式注射等量的生理鹽水。早期(缺血30min，，缺血再灌注后2h,4h，6h)檢測(cè)各組實(shí)驗(yàn)動(dòng)物血清及腦組織中的白細(xì)胞介素-1β（interleukinIL-1β）、IL-6、IL-10、腫瘤壞死因子-α（tumor necrosis factor-α, TNF-α）和腦組織內(nèi)炎性細(xì)胞，評(píng)價(jià)hUCB-MSCs移植調(diào)節(jié)炎性反應(yīng)的效果。神經(jīng)元凋亡、神經(jīng)生長(zhǎng)因子（nerve growth factor,NGF）、腦源性神經(jīng)營(yíng)養(yǎng)因子（brain derived neurotrophic factor, BDNF）、微管相關(guān)蛋白2（microtubule associated protein2，MAP2）、purdy評(píng)分的檢測(cè)用于評(píng)估hUCB-MSCs移植對(duì)實(shí)驗(yàn)動(dòng)物的神經(jīng)保護(hù)作用。 結(jié)果 缺血再灌注組和生理鹽水組血清中的IL-1β和IL-6在缺血后升高并在再灌注后2小時(shí)達(dá)到高峰，IL-10呈下降趨勢(shì)。MSC組IL-1β和IL-6在早期各時(shí)間點(diǎn)較缺血再灌注組和生理鹽水組明顯降低而IL-10顯著升高（P＜0.05）。第3d時(shí)腦組織勻漿檢測(cè)炎性因子也有相似的變化，且MSC組腦組織內(nèi)炎性細(xì)胞浸潤(rùn)情況較缺血再灌注組和生理鹽水組明顯減輕，浸潤(rùn)的炎性細(xì)胞數(shù)減少（P＜0.05）。 第3d時(shí)，缺血再灌注組和生理鹽水組腦組織中有大量凋亡細(xì)胞且凋亡指數(shù)（apoptosis index, AI）較高，而MSC組AI較缺血再灌注組和生理鹽水組明顯降低（P＜0.05）。MSC組血清NGF、BDNF含量較缺血再灌注組和生理鹽水組升高更為顯著（P0.05)而Purdy評(píng)分降低（P＜0.05）,缺血皮層區(qū)的MAP2含量亦增加（P0.05）。 結(jié)論 1、臍血間充質(zhì)干細(xì)胞移植可以調(diào)節(jié)兔腦缺血早期的炎性反應(yīng)。 2、臍血間充質(zhì)干細(xì)胞移植能抑制兔腦缺血皮層神經(jīng)元凋亡。 3、臍血間充質(zhì)干細(xì)胞移植能增加腦梗死兔血清中神經(jīng)營(yíng)養(yǎng)因子，促進(jìn)神經(jīng)元再生。 4、臍血間充質(zhì)干細(xì)胞移植能促進(jìn)兔腦缺血神經(jīng)功能缺損恢復(fù)。
[Abstract]:Objective to study the mechanism of early inflammatory response and neuroprotection after transplantation of human umbilical cord blood mesenchymal stem cells (human umbilical cord blood mesenchymal stemcells, hUCB-MSCs) in rabbits with focal cerebral ischemia-reperfusion injury. Methods umbilical cord blood was collected from 80 ml to 100 ml, hUCB-MSCs was isolated and cultured in vitro, and the cells cultured in the 6th passage were used for treatment and transplantation. 64 healthy New Zealand rabbits were randomly divided into three groups: sham operation group, ischemia reperfusion group, MSC group and saline group. The rabbit middle cerebral artery ischemia-reperfusion model was established by thread embolization. The (middle cerebral artery occlusion, MCAO), MSC group received 3mL5 脳 106 hUCB-MSCs suspensions through the femoral vein of the right leg, while the saline group received the same amount of normal saline in the same way. The levels of interleukin-1 尾 (interleukinIL-1 尾), IL-6,IL-10, tumor necrosis factor- 偽 (TNF- 偽) and inflammatory cells in brain tissue were detected at the early stage (30 min after ischemia and 4 h after reperfusion). The effect of hUCB-MSCs transplantation on the regulation of inflammatory response was evaluated. Neuronal apoptosis, nerve growth factor (nerve growth factor,NGF) and brain-derived neurotrophic factor (brain derived neurotrophic factor, BDNF),) microtubule-associated protein 2 (microtubule associated protein2,MAP2), purdy) score were used to evaluate the neuroprotective effect of hUCB-MSCs transplantation on experimental animals. Results the levels of IL-1 尾 and IL-6 in the serum of ischemia reperfusion group and normal saline group increased after ischemia and reached a peak at 2 hours after reperfusion, and IL-10 showed a decreasing trend. IL-1 尾 and IL-6 in MSC group were higher than those in ischemia reperfusion group at different time points. And normal saline group decreased significantly, but IL-10 increased significantly (P < 0. 05). On the 3rd day, the level of inflammatory cytokines in brain homogenate was similar, and the infiltration of inflammatory cells in MSC group was significantly less than that in ischemia reperfusion group and normal saline group (P < 0. 05), and the number of inflammatory cells in brain tissue homogenate was lower than that in ischemia reperfusion group and normal saline group (P < 0. 05). On the 3rd day, there were a large number of apoptotic cells in the brain tissues of ischemia reperfusion group and normal saline group, and the apoptotic index (apoptosis index, AI) was higher. However, AI in MSC group was significantly lower than that in ischemia reperfusion group and normal saline group (P < 0. 05). The serum NGF,BDNF content in MSC group was significantly higher than that in ischemia reperfusion group and normal saline group (P < 0. 05), and the Purdy score was decreased (P < 0. 05). The MAP2 content in ischemic cortex was also increased (P0.05). Conclusion 1. Umbilical cord blood mesenchymal stem cell transplantation can regulate the early inflammatory response of rabbit cerebral ischemia. 2. Umbilical cord blood mesenchymal stem cell transplantation can inhibit the apoptosis of rabbit cerebral ischemic cortical neurons. Transplantation of blood mesenchymal stem cells can increase neurotrophic factors in the serum of cerebral infarction rabbits. 4. Transplantation of mesenchymal stem cells from umbilical cord blood can promote the recovery of neural function defect after cerebral ischemia in rabbits.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R743

【參考文獻(xiàn)】

相關(guān)期刊論文 前8條

1 解建波,劉建民;瘦素、炎性細(xì)胞因子與腦梗死[J];國(guó)外醫(yī)學(xué).神經(jīng)病學(xué)神經(jīng)外科學(xué)分冊(cè);2003年05期

2 朱堯;汪青松;黃海麗;管葉明;;臍帶血間充質(zhì)干細(xì)胞改善兔腦梗死神經(jīng)生長(zhǎng)因子和神經(jīng)元凋亡的研究[J];中華老年心腦血管病雜志;2013年12期

3 王小蕊;侯相麟;席亞明;張海紅;;成人骨髓間充質(zhì)干細(xì)胞的鑒定與體外BrdU標(biāo)記[J];中國(guó)組織工程研究與臨床康復(fù);2008年21期

4 孫國(guó)軍;趙文靜;陳曦;葉冬霞;李晶;;臍血間充質(zhì)干細(xì)胞的分離培養(yǎng)及生物學(xué)特性[J];中國(guó)組織工程研究與臨床康復(fù);2011年14期

5 許二赫;武劍;賈建平;;血管性癡呆患者血漿中Aβ及Tau蛋白的檢測(cè)分析[J];中風(fēng)與神經(jīng)疾病雜志;2010年05期

6 洪小楊,王斌,杜江,朱德新,封志純;人臍帶血間充質(zhì)干細(xì)胞體外分離培養(yǎng)研究[J];中國(guó)誤診學(xué)雜志;2005年17期

7 ;Bone marrow-derived mesenchymal stem cells protect against experimental liver fibrosis in rats[J];World Journal of Gastroenterology;2005年22期

8 張侃;王洪新;閆玉仙;唐江偉;高路燕;魏中平;;長(zhǎng)春西汀對(duì)大鼠腦缺血再灌注損傷炎性反應(yīng)機(jī)制的影響[J];中國(guó)神經(jīng)免疫學(xué)和神經(jīng)病學(xué)雜志;2012年06期

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