【摘要】：目的:觀察電針"長(zhǎng)強(qiáng)"穴對(duì)FMR1基因敲除小鼠學(xué)習(xí)記憶能力及海馬區(qū)NLGN-3蛋白表達(dá)的影響。方法:(1)將FMR1基因敲除小鼠純合子一雌一雄合籠繁殖,其所產(chǎn)幼鼠于19日齡進(jìn)行分籠、標(biāo)記、采血,利用PCR法對(duì)其進(jìn)行基因鑒定,符合條件的FMR1基因敲除小鼠于23日齡隨機(jī)分為長(zhǎng)強(qiáng)組、非穴組、模型組,每組10只,共30只。長(zhǎng)強(qiáng)組取"長(zhǎng)強(qiáng)"穴并用電針干預(yù),干預(yù)過程采用30號(hào)0.5寸毫針,進(jìn)針深度0.3寸,波型為連續(xù)波,頻率為2 Hz,強(qiáng)度為2mA,持續(xù)20min,連續(xù)治療6d為1個(gè)療程,共2個(gè)療程,療程間間隔1d;非穴組取小鼠右助弓最低點(diǎn)上1cm處作為非經(jīng)非穴點(diǎn)并用電針干預(yù),余操作同長(zhǎng)強(qiáng)組;模型組在相同環(huán)境下飼養(yǎng),每日只進(jìn)行相同的網(wǎng)兜固定,不采取其它任何干預(yù)。(2)所有分組于23日齡及41日齡進(jìn)行Morris水迷宮測(cè)試,分別檢測(cè)小鼠干預(yù)前后的學(xué)習(xí)、記憶能力,于46日齡采用免疫組化法檢測(cè)小鼠海馬區(qū)NLGN-3蛋白的表達(dá)情況。(3)統(tǒng)計(jì)方法:利用SPSS20.0統(tǒng)計(jì)軟件對(duì)實(shí)驗(yàn)數(shù)據(jù)進(jìn)行單因素方差分析。結(jié)果:(1)水迷宮實(shí)驗(yàn)結(jié)果:①干預(yù)前逃避潛伏期分別為:長(zhǎng)強(qiáng)組(62.04±6.96)s、非穴組(55.46±9.81)s、模型組(57.83±17.72)s,三組組間比較,無顯著性差異(P0.05);②干預(yù)后逃避潛伏期分別為:長(zhǎng)強(qiáng)組(30.74±7.88)s、非穴組(40.82±8.05)s、模型組(43.63±12.27)s,長(zhǎng)強(qiáng)組與非穴組、模型組比較,差異具有顯著性(P0.05);③干預(yù)前原平臺(tái)象限時(shí)間/總時(shí)間分別為:長(zhǎng)強(qiáng)組(0.14±0.05)、非穴組(0.17±0.08)、模型組(0.16±0.07),三組組間比較,無顯著性差異(P0.05);④干預(yù)后原平臺(tái)象限時(shí)間/總時(shí)間分別為:長(zhǎng)強(qiáng)組(0.31±0.06)、非穴組(0.21±0.06)、模型組(0.17±0.04),長(zhǎng)強(qiáng)組與非穴組、模型組比較,差異具有顯著性(P0.05);(2)免疫組化法結(jié)果:干預(yù)后海馬區(qū)NLGN-3蛋白的灰度值分別為:長(zhǎng)強(qiáng)組(179.32±13.48)、非穴組(196.69±11.80)、模型組(197.615±7.99),長(zhǎng)強(qiáng)組與非穴組、模型組比較,差異具有顯著性(P0.05)。結(jié)論:電針"長(zhǎng)強(qiáng)"穴能提高FMR1基因敲除小鼠的學(xué)習(xí)記憶能力,其機(jī)制可能與促進(jìn)海馬區(qū)NLGN-3蛋白表達(dá)有關(guān)。
[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 鹵6.96) s in the long strong group, (55.46 鹵9.81) s in the non-point group and (57.83 鹵17.72) s in the model group. There was no significant difference (P0.05). 2Escape latency after intervention was (30.74 鹵7.88) s in the long strong group, (40.82 鹵8.05) s in the non-point group, (43.63 鹵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 鹵0.05), (0.17 鹵0.08) and (0.16 鹵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 鹵0.06), (0.21 鹵0.06), (0.17 鹵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 鹵13.48), (196.69 鹵11.80), (197.615 鹵7.99) in the long intensity group, (196.69 鹵11.80) and (197.615 鹵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.
【學(xué)位授予單位】：福建中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R245

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