本文選題：骨髓間充質(zhì)干細(xì)胞　切入點(diǎn)：腦缺血/再灌注　出處：《吉林大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:本實(shí)驗(yàn)主要探討骨髓間充質(zhì)干細(xì)胞對發(fā)生腦缺血/再灌注損傷的大鼠,在腦梗死體積、神經(jīng)功能損傷評分、腦缺血區(qū)域細(xì)胞的凋亡情況等方面的影響,并對其通過抑制神經(jīng)細(xì)胞凋亡,從而對神經(jīng)細(xì)胞發(fā)揮保護(hù)作用的可能機(jī)制給予研究。方法:對大鼠骨髓進(jìn)行初步分離,細(xì)胞培養(yǎng),骨髓間充質(zhì)干細(xì)胞鑒定,最終標(biāo)記等步驟,從而獲取實(shí)驗(yàn)用骨髓間充質(zhì)干細(xì)胞;將SD雄性大鼠60只進(jìn)行分組,隨機(jī)平均分為模型組、實(shí)驗(yàn)組、假手術(shù)組,共設(shè)3組,20只每組。假手術(shù)組不行栓塞大腦中動脈處理,其余組利用線栓栓塞大鼠大腦中動脈,缺血90分鐘后,取出線栓,造成大鼠腦缺血再灌注,從而建立大鼠腦缺血再灌注模型,再灌注時(shí)間為24 h。實(shí)驗(yàn)組給予濃度為1×109/L的骨髓間充質(zhì)干細(xì)胞1ml,采用尾靜脈注射法,另兩組經(jīng)尾靜脈給予1ml生理鹽水。分別在術(shù)后第1 Day、第1Week、第2 Weeks進(jìn)行神經(jīng)功能評分;在術(shù)后第2周測定腦梗死體積;RT-PCR法及Western blot檢測Bax/Bcl-2的m RNA和蛋白比例;檢測caspase-3蛋白含量、檢測Survivin蛋白表達(dá)情況。結(jié)果:與模型組比較,實(shí)驗(yàn)組神經(jīng)功能評分顯著降低(P0.05),腦梗死體積顯著減小(P0.05),Bcl-2的m RNA和蛋白表達(dá)明顯升高;而Bax的m RNA和蛋白表達(dá)明顯降低(P0.05),caspase-3蛋白表達(dá)減少(P0.05),Survivin蛋白表達(dá)增加(P0.05)。結(jié)論:1、骨髓間充質(zhì)干細(xì)胞具有保護(hù)神經(jīng)細(xì)胞的作用,可以減少腦梗死體積。2、骨髓間充質(zhì)干細(xì)胞能夠減輕缺血灌注后神經(jīng)功能的損傷,促進(jìn)神經(jīng)功能的恢復(fù)。3、上調(diào)Bcl-2m RNA的表達(dá),促進(jìn)Bcl-2蛋白表達(dá)。4.降低Baxm RNA的表達(dá),減少Bax蛋白含量。5.骨髓間充質(zhì)干細(xì)胞能夠增加Survivin蛋白的表達(dá),減少caspase-3蛋白的表達(dá)。綜上所述,骨髓間充質(zhì)干細(xì)胞能抑制神經(jīng)細(xì)胞凋亡,提高神經(jīng)細(xì)胞對缺血再灌注損傷的耐受力,發(fā)揮神經(jīng)保護(hù)作用�？傊撬栝g充質(zhì)干細(xì)胞能夠有效抑制缺血再灌注后神經(jīng)細(xì)胞的凋亡,但其更為詳細(xì)的機(jī)制仍需進(jìn)一步的深入研究。
[Abstract]:Objective: to investigate the effects of bone marrow mesenchymal stem cells (BMSCs) on cerebral infarction volume, neurological function injury score, apoptosis in ischemic area of brain in rats with cerebral ischemia / reperfusion injury. The possible mechanism of its protective effect on nerve cells was studied by inhibiting neuronal apoptosis. Methods: the primary isolation of rat bone marrow, cell culture, identification of bone marrow mesenchymal stem cells, and final labeling, etc. In order to obtain bone marrow mesenchymal stem cells for experiment, 60 male SD rats were divided into three groups: model group, experimental group and sham operation group. In the other group, the middle cerebral artery was embolized with thread embolism. After 90 minutes of ischemia, the thread embolus was removed to make the cerebral ischemia-reperfusion in rats, and the model of cerebral ischemia-reperfusion was established. The time of reperfusion was 24 hours. The experimental group was given 1 ml of bone marrow mesenchymal stem cells (1 脳 10 9 / L) by caudal vein injection, and the other two groups were given 1 ml of normal saline through tail vein. The neurological function was evaluated at 1 day, 1 week and 2 Weeks after operation. At the second week after operation, the ratio of m RNA and protein of Bax/Bcl-2 was detected by RT-PCR and Western blot, and the content of caspase-3 protein and the expression of Survivin protein were detected. Results: compared with the model group, the ratio of m RNA and protein of Bax/Bcl-2 was detected by RT-PCR and Western blot at the second week after operation. In the experimental group, the scores of nerve function decreased significantly (P 0.05), and the volume of cerebral infarction significantly decreased the expression of m RNA and protein of Bcl 2. The expression of m RNA and protein in Bax decreased significantly, while the expression of caspase-3 decreased significantly. Conclusion: bone marrow mesenchymal stem cells (BMSCs) can protect nerve cells by increasing the expression of survivin. Bone marrow mesenchymal stem cells can reduce the injury of nerve function, promote the recovery of nerve function, up-regulate the expression of Bcl-2m RNA, promote the expression of Bcl-2 protein, and decrease the expression of Baxm RNA. Bone marrow mesenchymal stem cells can increase the expression of Survivin protein and decrease the expression of caspase-3 protein. In conclusion, bone marrow mesenchymal stem cells can inhibit neuronal apoptosis. In short, bone marrow mesenchymal stem cells can effectively inhibit neuronal apoptosis after ischemia-reperfusion injury. However, its more detailed mechanism still needs to be further studied.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R743.3

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