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电针对高血压大鼠脑梗死后神经前体细胞移行的实验性研究

Electroacupuncture Experimental Studies of Cell Migration of Neural Precursor Cells in the Hypertensive Rats After Acute Cerebral Infarction

【作者】 王少帅

【导师】 李常新;

【作者基本信息】 山西医科大学 , 神经病学, 2012, 硕士

【摘要】 目的:脑梗死后功能恢复是一个非常复杂的过程,电针在脑梗死康复治疗中有显著疗效,但是具体机制尚不明确。神经发生在神经损伤恢复中起着一定作用,电针治疗是否与神经发生有关是启动本课题的关键问题。本文旨在探讨成年高血压大鼠脑梗死后电针作用机制及对内源性神经前体细胞移行的影响,指导临床治疗。方法:采用健康雄性,体重80~120g,鼠龄60~90天的SD大鼠,按已定型的双肾双夹法复制RHRSP模型,正常血压组不上银夹,术后每两周测一次血压。肾动脉狭窄术后16周,血压≥175mmHg且无脑卒中症状,体重450~500g的RHRSP;用电凝法制作MCAO模型。采取随机分组。假手术组:RHRSP非MCAO,1周、2周、3周、4周,每组3只,共12只;梗死组:RHRSP与MCAO模型成功1周、2周、3周、4周,每组6只,共24只;电针组:RHRSP与MCAO模型成功行电针治疗1周、2周、3周、4周,每组6只,共24只。各组均于梗死后行不同时间点进行神经功能评分,灌注取脑行病理切片,TTC染色,常规HE染色,免疫组织化学方法检测各组大鼠脑内病灶周围及海马区DCX、MMP-2阳性细胞表达。统计学方法采用SPSS17.0forWindows统计软件包进行统计分析。正态分布数据用均数±标准差进行描述,两组间的均数比较用t检验,多组间的均数比较用单因素方差分析,当方差分析具有统计学意义时进一步用SNK-q检验做两两比较,检验水平为α=0.05。结果:RHRSP后平均血压为198.37±21.45mmHg。按照18分综合评分法进行神经功能评分,假手术组评分18分(正常),MCAO后评分随时间逐渐增高,5d时恢复正常。电针后2d与3d大鼠神经功能评分高于梗死组(P<0.01);电针治疗后前3周梗死灶体积较梗死组缩小(P<0.05);DCX阳性细胞在脑梗死后灶周、海马表达增加,1周为高峰期;电针治疗1周末较梗死组增加(P<0.05),2~4周随时间延长数目较梗死组明显减少。MMP-2于脑梗死后1周时灶周表达明显增加,之后呈下降趋势,接近假手术组。电针治疗1周末较梗死组明显增加(P<0.05)。结论:高血压大鼠脑梗死后电针治疗能促进神经功能恢复,减小脑梗死体积;脑梗死可诱导神经前体细胞向缺血灶周围移行;急性期电针治疗可使缺血灶周移行的神经前体细胞增多,并且促进缺血灶周MMP-2的合成分泌,加快神经前体细胞移行,修复神经细胞缺损。

【Abstract】 Objective:Functional recovery after cerebral infarction is a very complex process, electricity has been recognized for neurological recovery after stroke, but the exact mechanism is not clear. Play a role in the recovery of the nerve damage due to neurogenesis, EA related to the occurrence of the nerve. This paper aims to investigate the migration of neural precursor cells (NPCs) and to examine the effect of electroacupuncture (EA) on them after acute cerebral infarction.Methods:Stroke-prone renovascular hypertensive rats (RHRSP) were made after the renal arteries in male SD rats weighting80-120g being constricted bilaterally with ring-shape silver clips. BP were measured once two weeks after the operation.16weeks after the operation, MCAO models were made by electric coagulation according Wahl’s method in RHRSP,which weighted450~500g with high BP (more than175mmHg) and without stroke signs. Take a random grouping.Sham operation group:RHRSP non-MCAO,1,2,3,4weeks, n=3, a total of12; cerebral infarction Group:RHRSP and the success of MCAO mode1,2,3,4weeks.n=6,24; EA group:RHRSP MCAO model successfully in electro-acupuncture treatment1,2,3,4weeks (n=6), a total of24. Each group in the infarction underwent neurological score, HE staining, the TTC staining. Paraffin sections immunohistochemical method for detection of brain at different time points around the lesion and the hippocampus DCX-positive cells of MMP-2expression, and cerebral infarction and control groups. Statistical methods using SPSS17.0for Windows statistical package for statistical analysis. Normal distribution data were expressed as x±s deviation are described between the two groups were compared by t-test groups using univariate analysis of variance, and further analysis of variance was statistically significant with SNK-q do a pairwise comparison test, the test level ofα=0.05.Results:RHRSP average blood pressure was198.37±21.45mmHg. In accordance with the neurological score of18points score, sham operation group to score18points (normal) score of MCAO after gradually increased over time, and5d back to normal.2d and3d neurological score higher than infarction after EA(P<0.01); the first three weeks after the infarct volume in the electro-acupuncture treatment than infarction group reduced(P<0.05); DCX-positive cells in the ischemic perifocal and hippocampus increase, one week for the peak period; EA treatment significantly increased on a weekend than infarction group(P<0.05),2to4weeks with time than infarction group was significantly reduced. Perifocal expression of MMP-2at1week after ischemia significantly increased, after a downward trend, close to the sham operation group. Electro-acupuncture treatment of a weekend than the infarction was significantly increased (P<0.05).Conclusion:The EA may promote the neurofunctional recovery, reduce infarct volume, NPCs to the infarct from around in cerebral ischemic area.To promote the synthesis and secretion of MMP-2, the acute phase is particularly evident, to accelerate the migration of neural precursor cells into the ischemic perifocal.

【关键词】 脑梗死电针DCXMMP-2神经前体细胞
【Key words】 StrokeElectroacupunctureDoublecortinMMP-2Neural precursor cell
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