【摘要】：背景:前期實(shí)驗(yàn)研究發(fā)現(xiàn),移植人羊膜間充質(zhì)干細(xì)胞能有效改善腦梗死大鼠的神經(jīng)損傷癥狀。目的:觀察移植人羊膜間充質(zhì)干細(xì)胞在大鼠腦梗死區(qū)域的存活、定植和分化情況。方法:將60只SD大鼠隨機(jī)分為3組,實(shí)驗(yàn)組、模型組采用線栓法制作大腦中動脈閉塞模型,假手術(shù)組只結(jié)扎血管,不插入線栓;造模后1 d,實(shí)驗(yàn)組于受損紋狀體和皮質(zhì)兩點(diǎn)原位植入10μL第3代人羊膜間充質(zhì)干細(xì)胞懸液(含細(xì)胞2×106/只),模型組與假手術(shù)組于相同部位注射等體積PBS。移植后1周連續(xù)監(jiān)測體質(zhì)量并進(jìn)行神經(jīng)缺損嚴(yán)重程度評分;移植后2周,TTC染色觀察腦梗死面積,蘇木精-伊紅染色觀察腦組織病理學(xué)變化,免疫熒光染色檢測移植大鼠大腦神經(jīng)細(xì)胞標(biāo)志物神經(jīng)元特異性核蛋白的表達(dá)。結(jié)果與結(jié)論:(1)體質(zhì)量與神經(jīng)缺損嚴(yán)重程度評分:與假手術(shù)組比較,模型組、實(shí)驗(yàn)組體質(zhì)量呈下降趨勢,實(shí)驗(yàn)組降低少于模型組,但差異無顯著性意義;模型組、實(shí)驗(yàn)組神經(jīng)功能缺損評分隨時(shí)間呈逐漸降低趨勢,但實(shí)驗(yàn)組評分明顯低于模型組;(2)腦梗死面積:模型組大腦皮質(zhì)出現(xiàn)局灶性缺血壞死且范圍較大,與模型組比較,實(shí)驗(yàn)組缺血灶面積明顯縮小;(3)腦組織病理:實(shí)驗(yàn)組梗死病灶范圍、神經(jīng)細(xì)胞及炎細(xì)胞浸潤均少于模型組;(4)免疫熒光染色:移植后1周,實(shí)驗(yàn)組腦組織可見較多的移植人羊膜間充質(zhì)干細(xì)胞,2周后可見移植的人羊膜間充質(zhì)干細(xì)胞向神經(jīng)樣細(xì)胞分化;(5)結(jié)果表明:人羊膜間充質(zhì)干細(xì)胞移植可在大鼠腦梗死區(qū)域定植、存活,且在原位能分化為神經(jīng)樣細(xì)胞。
[Abstract]:Background: transplantation of human amniotic mesenchymal stem cells (HMMSCs) can effectively improve the neurological injury symptoms in rats with cerebral infarction. Aim: to observe the survival, colonization and differentiation of transplanted human amniotic mesenchymal stem cells (HMMSCs) in cerebral infarction area of rats. Methods: sixty SD rats were randomly divided into three groups: the model group was made into middle cerebral artery occlusion model by thread occlusion, and the sham operation group only ligated the blood vessel without inserting the thread embolus. 1 day after modeling, the experimental group was implanted in situ with 10 渭 L human amniotic mesenchymal stem cell suspension (containing 2 脳 106 cells per mouse) at both the damaged striatum and cortex. The model group and the sham operation group were injected with PBS. of the same volume at the same site. One week after transplantation, the body mass was continuously monitored and the severity of nerve defect was evaluated, the area of cerebral infarction was observed by TTC staining and the pathological changes of brain tissue were observed by hematoxylin and eosin staining at 2 weeks after transplantation. The expression of neuron-specific nuclear protein was detected by immunofluorescence staining. Results and conclusion: (1) body mass and nerve defect severity score: compared with sham operation group, the body mass of model group, experimental group showed a downward trend, the experimental group decreased less than model group, but the difference was not significant, the model group, the model group, The neurological deficit score of the experimental group decreased gradually with time, but the score of the experimental group was obviously lower than that of the model group. (2) Cerebral infarction area: focal ischemic necrosis occurred in the cerebral cortex of the model group and the range was larger, compared with the model group. (3) brain histopathology: infarct area, nerve cell and inflammatory cell infiltration in experimental group were less than those in model group; (4) immunofluorescence staining: 1 week after transplantation, In the experimental group, more human amniotic mesenchymal stem cells could be found to differentiate into neural like cells 2 weeks after transplantation. (5) the results showed that the transplantation of human amniotic mesenchymal stem cells could be colonized in the infarcted area of rat brain. Survival, and in situ differentiation into neuronal cells.
【作者單位】： 遵義醫(yī)學(xué)院附屬醫(yī)院貴州省細(xì)胞工程重點(diǎn)實(shí)驗(yàn)室;遵義醫(yī)學(xué)院藥學(xué)院;遵義醫(yī)學(xué)院附屬醫(yī)院臨床醫(yī)學(xué)研究所;遵義醫(yī)學(xué)院教育部基礎(chǔ)藥理學(xué)重點(diǎn)實(shí)驗(yàn)室;
【基金】：貴州省科學(xué)技術(shù)基金項(xiàng)目資助(黔教研合J字LKZ[2011]24號)~~
【分類號】：R743.33

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