本文選題：載脂蛋白J(ApoJ) + 腦出血��； 參考：《西南醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:探討攜帶ApoJ基因的骨髓間充質(zhì)干細(xì)胞對(duì)大鼠實(shí)驗(yàn)性腦出血的治療作用。方法:采取脂質(zhì)體介導(dǎo)用重組質(zhì)粒pEGFP-N1-apoJ轉(zhuǎn)染體外培養(yǎng)的骨髓間充質(zhì)干細(xì)胞(BMSCs),分別于轉(zhuǎn)染后24,48,72,96h在熒光顯微鏡下觀察相同體積脂質(zhì)體介導(dǎo)不同重組質(zhì)粒量的轉(zhuǎn)染效果,篩選出轉(zhuǎn)染率最高的轉(zhuǎn)染組。將102只雄性SD大鼠隨機(jī)分為轉(zhuǎn)染組、BMSCs組和NaCl組。每組再分為24h,48h,72h,96h四個(gè)亞組。利用腦立體定位儀建立大鼠腦出血模型,三組分別于造模24h后顱內(nèi)注射等體積的轉(zhuǎn)染細(xì)胞懸液、BMSCs懸液和生理鹽水,分別于處理后24h,48h,72h,96h處死各對(duì)應(yīng)亞組大鼠。進(jìn)行各時(shí)間點(diǎn)改良神經(jīng)功能評(píng)分(mNSS);用干濕質(zhì)量法測(cè)量腦組織含水量;用蛋白免疫印跡法檢測(cè)載脂蛋白-J在各時(shí)間點(diǎn)的表達(dá)。結(jié)果:1.等體積脂質(zhì)體條件下,四個(gè)細(xì)胞轉(zhuǎn)染組(重組質(zhì)粒量分別為0.8、1.2、1.6、2.0)在72h時(shí)轉(zhuǎn)染率高于同組在24h,48h,96h的轉(zhuǎn)染率(P均0.05);且與另外三個(gè)細(xì)胞轉(zhuǎn)染組(重組質(zhì)粒量分別為0.8、1.2、2.0)比較,重組質(zhì)粒量為1.6的細(xì)胞轉(zhuǎn)染組在72h時(shí)熒光表達(dá)最強(qiáng),轉(zhuǎn)染效率高于同時(shí)間點(diǎn)的其他三組(P均0.05);2.mNSS結(jié)果顯示,轉(zhuǎn)染組在實(shí)驗(yàn)干預(yù)后1,3,5,7d得分顯著低于同時(shí)間點(diǎn)的BMSCs組與NaCl組(P均0.05);3.腦組織含水量測(cè)定結(jié)果顯示,轉(zhuǎn)染組在1,3,5,7d腦組織含水量低于同時(shí)間點(diǎn)的BMSCs組與NaCl組(P均0.05);4.Western blot結(jié)果顯示,與BMSCs組與NaCl組比較,轉(zhuǎn)染組腦組織內(nèi)ApoJ表達(dá)水平均高于同時(shí)間點(diǎn)的兩組(P均0.05)。結(jié)論:攜帶ApoJ基因的骨髓間充質(zhì)干細(xì)胞能夠在ICH大鼠腦組織內(nèi)表達(dá)載脂蛋白J,外源性ApoJ對(duì)腦出血具有治療作用,可以促進(jìn)腦出血大鼠的神經(jīng)功能的恢復(fù),減輕腦水腫,改善腦出血大鼠預(yù)后。
[Abstract]:Aim: to investigate the therapeutic effect of bone marrow mesenchymal stem cells carrying ApoJ gene on experimental intracerebral hemorrhage in rats. Methods: the recombinant plasmid pEGFP-N1-apoJ was transfected into bone marrow mesenchymal stem cells (BMSCs) by liposome-mediated transfection in vitro. The transfection effects of different amount of recombinant plasmids mediated by the same volume of liposome were observed under fluorescence microscope at 24: 48 ~ 72 ~ 96 h after transfection. The transfection group with the highest transfection efficiency was selected. 102 male SD rats were randomly divided into BMSCs group and NaCl group. Each group was subdivided into four subgroups: 24 h, 48 h, 72 h and 96 h. The intracerebral hemorrhage model of rats was established by brain stereotactic locator. BMSCs suspension and normal saline were injected into the brain of the three groups 24 hours after the establishment of the model. The rats of the corresponding subgroups were killed at 24 h, 48 h, 72 h and 96 h, respectively. The modified neurologic function scores were evaluated at each time point, the water content of brain tissue was measured by wet and dry mass method, and the expression of apolipoprotein J at each time point was detected by Western blot. The result is 1: 1. In the same volume liposome condition, the transfection efficiency of the four cell transfection groups (the amount of recombinant plasmids was 0.81.21.62.0) at 72 h was higher than that in the same group at 24 h, 48 h or 96 h, and was compared with the other three cell transfection groups (the amount of recombinant plasmids was 0.88 1.22 2.0). The transfection group with 1.6 recombinant plasmid had the strongest fluorescence expression at 72 h, and the transfection efficiency was higher than that of the other three groups at the same time point (P 0.05) and 2.mNSS. The results showed that the score of transfection group was significantly lower than that of BMSCs group and NaCl group on the 1st and 5th day after experimental intervention. The water content of brain tissue in transfection group was lower than that in BMSCs group and NaCl group at 1 ~ 3 ~ 5 ~ 5 ~ 5 ~ 7 days after transfection. 4. The results showed that the water content of brain tissue in transfection group was lower than that in BMSCs group and NaCl group at the same time point. 4. The results showed that compared with BMSCs group and NaCl group. The expression of ApoJ in brain tissue of the transfected group was higher than that of the two groups at the same time point (P < 0.05). Conclusion: bone marrow mesenchymal stem cells carrying ApoJ gene can express apolipoprotein J in the brain of ICH rats. Exogenous ApoJ can promote the recovery of neural function and alleviate cerebral edema in ICH rats. To improve the prognosis of rats with intracerebral hemorrhage.
【學(xué)位授予單位】：西南醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R743.34

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