电针“长强”穴对FMR1基因敲除小鼠海马区NLGN-3蛋白表达的影响

发布时间：2018-11-21 10:02

【摘要】：目的:观察电针"长强"穴对FMR1基因敲除小鼠学习记忆能力及海马区NLGN-3蛋白表达的影响。方法:(1)将FMR1基因敲除小鼠纯合子一雌一雄合笼繁殖,其所产幼鼠于19日龄进行分笼、标记、采血,利用PCR法对其进行基因鉴定,符合条件的FMR1基因敲除小鼠于23日龄随机分为长强组、非穴组、模型组,每组10只,共30只。长强组取"长强"穴并用电针干预,干预过程采用30号0.5寸毫针,进针深度0.3寸,波型为连续波,频率为2 Hz,强度为2mA,持续20min,连续治疗6d为1个疗程,共2个疗程,疗程间间隔1d;非穴组取小鼠右助弓最低点上1cm处作为非经非穴点并用电针干预,余操作同长强组;模型组在相同环境下饲养,每日只进行相同的网兜固定,不采取其它任何干预。(2)所有分组于23日龄及41日龄进行Morris水迷宫测试,分别检测小鼠干预前后的学习、记忆能力,于46日龄采用免疫组化法检测小鼠海马区NLGN-3蛋白的表达情况。(3)统计方法:利用SPSS20.0统计软件对实验数据进行单因素方差分析。结果:(1)水迷宫实验结果:①干预前逃避潜伏期分别为:长强组(62.04±6.96)s、非穴组(55.46±9.81)s、模型组(57.83±17.72)s,三组组间比较,无显著性差异(P0.05);②干预后逃避潜伏期分别为:长强组(30.74±7.88)s、非穴组(40.82±8.05)s、模型组(43.63±12.27)s,长强组与非穴组、模型组比较,差异具有显著性(P0.05);③干预前原平台象限时间/总时间分别为:长强组(0.14±0.05)、非穴组(0.17±0.08)、模型组(0.16±0.07),三组组间比较,无显著性差异(P0.05);④干预后原平台象限时间/总时间分别为:长强组(0.31±0.06)、非穴组(0.21±0.06)、模型组(0.17±0.04),长强组与非穴组、模型组比较,差异具有显著性(P0.05);(2)免疫组化法结果:干预后海马区NLGN-3蛋白的灰度值分别为:长强组(179.32±13.48)、非穴组(196.69±11.80)、模型组(197.615±7.99),长强组与非穴组、模型组比较,差异具有显著性(P0.05)。结论:电针"长强"穴能提高FMR1基因敲除小鼠的学习记忆能力,其机制可能与促进海马区NLGN-3蛋白表达有关。
[Abstract]:Aim: to observe the effect of electroacupuncture on the learning and memory ability of FMR1 knockout mice and the expression of NLGN-3 protein in hippocampus. Methods: (1) the homozygotes of FMR1 knockout mice, female and male, were divided into cages, labeled, and blood samples were collected at 19 days of age, and their genes were identified by PCR method. Eligible FMR1 knockout mice were randomly divided into Changqiang group, non-acupoint group and model group at the age of 23 days, with 10 mice in each group, 30 mice in each group. In the Changqiang group, the points of "Changqiang" were taken and electroacupuncture was used to intervene. The intervention process was 30 0.5 inch millimeter needle, the depth of the needle was 0.3 inch, the wave type was continuous wave, the frequency was 2 Hz, intensity 2 Ma, lasted for 20 minutes, and the treatment lasted for 6 days as a course of treatment. There were 2 courses of treatment, the interval between the two courses was 1 day; In the non-acupoint group, the 1cm at the lowest point of the right auxiliary arch was taken as the non-meridian non-acupoint point and electroacupuncture intervention, and the remaining operation was in the same long and strong group. The model group was fed in the same environment, only fixed with the same net pocket daily without any other intervention. (2) Morris water maze test was performed in all groups at 23 and 41 days of age, and the learning of mice before and after intervention was measured. At 46 days of age, the expression of NLGN-3 protein in hippocampus of mice was detected by immunohistochemical method. (3) Statistical method: univariate ANOVA was performed with SPSS20.0 software. Results: (1) the results of water maze test: 1the escape latency before intervention was (62.04 卤6.96) s in the long strong group, (55.46 卤9.81) s in the non-point group and (57.83 卤17.72) s in the model group. There was no significant difference (P0.05). 2Escape latency after intervention was (30.74 卤7.88) s in the long strong group, (40.82 卤8.05) s in the non-point group, (43.63 卤12.27) s in the model group, and there was significant difference between the model group and the non-point group (P0.05). 3before intervention, the original platform quadrant time / total time were (0.14 卤0.05), (0.17 卤0.08) and (0.16 卤0.07), respectively. There was no significant difference among the three groups (P0.05). 4After intervention, the original platform quadrant time / total time were (0.31 卤0.06), (0.21 卤0.06), (0.17 卤0.04), respectively. The difference was significant (P0.05). (2) Immunohistochemical results: the gray values of NLGN-3 protein in hippocampus after intervention were (179.32 卤13.48), (196.69 卤11.80), (197.615 卤7.99) in the long intensity group, (196.69 卤11.80) and (197.615 卤7.99) in the model group, respectively. The difference was significant (P0.05). Conclusion: electroacupuncture at "Changqiang" can improve the learning and memory ability of FMR1 knockout mice, and its mechanism may be related to the promotion of NLGN-3 protein expression in hippocampus.
【学位授予单位】：福建中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R245

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