本文選題：電針 + 異氟醚�。� 參考：《廣州中醫(yī)藥大學(xué)》2014年碩士論文

【摘要】：目的： 探討電針對(duì)異氟醚誘導(dǎo)的AD樣病神經(jīng)毒性的影響及作用機(jī)制。 方法： 阿爾茨海默病APPswe/PS1dE9雙轉(zhuǎn)基因小鼠(AD小鼠)及野生型小鼠各24只,隨機(jī)分為AD小鼠對(duì)照組(T-Con)、AD小鼠異氟醚組(T-Iso)、AD小鼠電針組(T-EA)、野生小鼠對(duì)照組(W-Con)、野生小鼠異氟醚組(W-Iso)及野生小鼠電針組(W-EA),n=8。異氟醚暴露前電針組進(jìn)行電針處理,選取百會(huì)、涌泉穴,每次15min,1次／天,連續(xù)3天；對(duì)照組和異氟醚組只固定不針刺。麻醉暴露：Iso組和EA組由30%氧氣和70%氮?dú)饣旌蠚怏w輸送異氟醚通過麻醉箱行麻醉暴露,調(diào)整麻醉箱內(nèi)異氟醚濃度至1.2%并維持穩(wěn)定；Con組同時(shí)置于另外相同麻醉箱內(nèi),吸入30%氧氣和70%氮?dú)饣旌蠚怏w4h。Morris水迷宮測(cè)試小鼠學(xué)習(xí)記憶能力的變化；TUNEL檢測(cè)海馬CA1區(qū)細(xì)胞凋亡變化；免疫組化檢測(cè)各組海馬CA1區(qū)激活型caspase-3、Aβ的表達(dá)；Western blot檢測(cè)海馬APP-C83及APP-C99的表達(dá)。 結(jié)果：一、行為學(xué)測(cè)試 (一)AD小鼠與野生學(xué)習(xí)及記憶能力比較：各組小鼠的逃避潛伏期隨著測(cè)試天數(shù)的增加逐漸縮短；AD小鼠的逃避潛伏期長(zhǎng)于野生小鼠,穿越平臺(tái)次數(shù)及第一象限停留時(shí)間百分比低于野生小鼠,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。 (二)AD小鼠在水迷宮測(cè)試中的成績(jī)：定位航行測(cè)試中逃避潛伏期比較：T-ConT-EAT-Iso, T-Iso與T-Con之間有顯著性差異(P0.05),T-EA與T-Iso間差異有統(tǒng)計(jì)學(xué)意義(P0.05)�？臻g探索實(shí)驗(yàn)比較：第一象限停留時(shí)間百分比T-ConT-EAT-Iso,T-Iso與T-Con差異有統(tǒng)計(jì)學(xué)意義(P0.05),T-EA與T-Iso差異有統(tǒng)計(jì)學(xué)意義(P0.05)；AD小鼠60s內(nèi)穿越平臺(tái)次數(shù)T-ConT-EAT-Iso, T-Iso及T-EA與T-Con差異有統(tǒng)計(jì)學(xué)意義(P0.05)。 (三)野生小鼠在水迷宮測(cè)試中的成績(jī)：定位航行測(cè)試中逃避潛伏期比較：W-ConW-EAW-Iso,W-Iso與W-Con之間有顯著性差異(P0.05),W-EA與W-Iso間差異有統(tǒng)計(jì)學(xué)意義(P0.05)�？臻g探索實(shí)驗(yàn)比較：野生組小鼠在第一象限停留時(shí)間百分比及60s內(nèi)穿越平臺(tái)次數(shù)比較：W-ConW-EAW-Iso,三組之間差異無統(tǒng)計(jì)學(xué)意義(P0.05)。 二、激活型caspase-3的表達(dá)及TUNEL細(xì)胞凋亡情況比較 (一)AD小鼠：AD小鼠海馬區(qū)caspase-3陽(yáng)性細(xì)胞數(shù)目比較：T-ConT-EAT-Iso,T-Iso與T-Con間差異統(tǒng)計(jì)學(xué)意義(P0.05), T-EA與T-Iso間差異有統(tǒng)計(jì)學(xué)意義(P0.05)。AD小鼠海馬區(qū)TUNEL細(xì)胞凋亡情況與激活型caspase-3的表達(dá)相一致。 (二)野生小鼠：野生小鼠海馬區(qū)caspase-3陽(yáng)性細(xì)胞數(shù)目比較：W-ConW-EAW-Iso,三組之間差異無統(tǒng)計(jì)學(xué)意義(P0.05)。野生小鼠海馬區(qū)TUNEL細(xì)胞凋亡情況與激活型caspase-3的表達(dá)相符。 三、APP-C83、APP-C99及Aβ表達(dá)情況 (一) Western blot檢測(cè)APP-C83及APP-C99的表達(dá)：APP-C83/GAPDH:T-Iso組APP-C83的表達(dá)低于T-Con和W-Con (P0.05); T-EA組APP-C83的表達(dá)高于T-Iso組(P0.05). APP-C83/GAPDH:T-Iso組APP-C99的表達(dá)高于T-Con和W-Con; T-EA組APP-C99的表達(dá)低于T-Iso組(P0.05)。 (二)免疫組化檢測(cè)Aβ的表達(dá)：6月齡AD小鼠海馬區(qū)已有明顯的老年斑形成；T-Iso及T-EA組Aβ表達(dá)高于T-Con組(P0.05)；T-EA組A β表達(dá)低于T-Iso組,但兩組之間差異無統(tǒng)計(jì)學(xué)意義(P0.05)。 結(jié)論： 一、6月齡AD小鼠的學(xué)習(xí)記憶能力較野生小鼠已經(jīng)有明顯下降。 二、電針可在一定程度上減輕異氟醚誘使的AD小鼠的認(rèn)知功能損害。 三、電針可通過抑制異氟醚引起的AD小鼠海馬區(qū)激活型caspase-3的過度表達(dá),減少神經(jīng)元凋亡,減輕異氟醚對(duì)AD小鼠造成的神經(jīng)細(xì)胞損害。 四、電針可抑制異氟醚導(dǎo)致的APP-C99的過量表達(dá),一定程度上減少Aβ的生成和聚集。
[Abstract]:Objective:
Objective to investigate the effect and mechanism of Electroacupuncture on isoflurane induced AD like neurotoxicity.
Method:
24 APPswe/PS1dE9 double transgenic mice (AD mice) and 24 wild type mice were randomly divided into AD mice control group (T-Con), AD mice isoflurane group (T-Iso), AD mice electroacupuncture group (T-EA), wild mouse control group (W-Con), wild mice isoflurane group (W-Iso) and wild mice electroacupuncture group (W-EA), before exposure to isoflurane exposure electroacupuncture group Electroacupuncture treatment, selected Baihui, Yongquan point, each time 15min, 1 times per day, 3 days, the control group and isoflurane group only fixed no acupuncture. Anesthesia exposure: Iso and EA group of 30% oxygen and 70% nitrogen gas mixture of isoflurane through anaesthesia box exposure, adjust the concentration of isoflurane in the drunken box to 1.2% and maintain stability; Con group In the same anaesthesia box, the changes of learning and memory ability of mice were measured by inhaling 30% oxygen and 70% nitrogen gas mixture 4h.Morris water maze; TUNEL was used to detect the changes in the apoptosis of hippocampal CA1 area; immunohistochemistry was used to detect the expression of Caspase-3, A beta in hippocampus CA1 region, and Western blot to detect the expression of APP-C83 and APP-C99 in hippocampus.
Results: first, behavioral test
(1) compared with the learning and memory ability of AD mice, the escape latency of mice in each group gradually shortened with the increase of test days; the escape latency of AD mice was longer than that of wild mice, and the number of crossing platform and the percentage of first quadrant residence time were lower than those in the wild mice, and the difference was statistically significant (P0.05).
(two) the results of AD mice in the water maze test: the escape latency of the navigation test was compared: T-ConT-EAT-Iso, T-Iso and T-Con had significant difference (P0.05), the difference between T-EA and T-Iso was statistically significant (P0.05). The comparison of the space exploration experiment: the percentage T-ConT-EAT-Iso in the first quadrant and the difference between T-Iso and T-Con The difference between T-EA and T-Iso was statistically significant (P0.05), and the number of T-ConT-EAT-Iso, T-Iso and T-EA and T-Con of 60s in AD mice were statistically significant (P0.05), and the difference between T-Iso and T-Iso in AD mice was statistically significant.
(three) the results of wild mice in the water maze test: W-ConW-EAW-Iso, W-Iso and W-Con have significant difference (P0.05), there is a significant difference between W-EA and W-Iso (P0.05). The comparison of space exploration experiment: the percentage of the first quadrant residence time in the wild group and the passing through of 60s in the wild group There was no significant difference between the three groups (W-ConW-EAW-Iso): P0.05.
Two, the expression of activated caspase-3 and TUNEL cell apoptosis were compared.
(1) AD mice: the number of Caspase-3 positive cells in the hippocampus of AD mice was compared: T-ConT-EAT-Iso, the difference between T-Iso and T-Con was statistically significant (P0.05), and there was a significant difference between T-EA and T-Iso (P0.05) the apoptosis of TUNEL cells in the hippocampus of.AD mice was in accordance with the expression of activated type.
(two) wild mice: the number of Caspase-3 positive cells in the hippocampus of wild mice was compared: W-ConW-EAW-Iso, there was no significant difference between the three groups (P0.05). The apoptosis of the TUNEL cells in the hippocampal region of the wild mice was in accordance with the expression of the activated caspase-3.
Three, APP-C83, APP-C99 and A beta expression
(1) the expression of APP-C83 and APP-C99 by Western blot: the expression of APP-C83 in APP-C83/GAPDH:T-Iso group was lower than that of T-Con and W-Con (P0.05), and the expression of APP-C83 in T-EA group was higher than that in T-Iso group.
(two) immunohistochemical detection of the expression of A beta: 6 month old AD mice had obvious senile plaque formation; A beta expression in T-Iso and T-EA groups was higher than that of T-Con group (P0.05), and A beta expression in T-EA group was lower than that of T-Iso group, but there was no statistical difference between the two groups (P0.05).
Conclusion:
The learning and memory ability of one, 6 month old AD mice decreased significantly compared with that of wild mice.
Two, EA can alleviate the cognitive impairment of AD mice induced by isoflurane to a certain extent.
Three, electroacupuncture can inhibit the overexpression of activated caspase-3 in the hippocampal region of AD mice induced by isoflurane, reduce neuronal apoptosis and reduce the nerve cell damage caused by isoflurane in AD mice.
Four, EA can inhibit the overexpression of APP-C99 induced by isoflurane, and reduce the production and accumulation of A beta to some extent.
【學(xué)位授予單位】：廣州中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R245

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