电针对异氟醚诱导的AD样病神经毒性的影响

发布时间：2018-06-22 18:41

本文选题：电针 + 异氟醚　；参考：《广州中医药大学》2014年硕士论文

【摘要】：目的：探讨电针对异氟醚诱导的AD样病神经毒性的影响及作用机制。方法：阿尔茨海默病APPswe/PS1dE9双转基因小鼠(AD小鼠)及野生型小鼠各24只,随机分为AD小鼠对照组(T-Con)、AD小鼠异氟醚组(T-Iso)、AD小鼠电针组(T-EA)、野生小鼠对照组(W-Con)、野生小鼠异氟醚组(W-Iso)及野生小鼠电针组(W-EA),n=8。异氟醚暴露前电针组进行电针处理,选取百会、涌泉穴,每次15min,1次／天,连续3天；对照组和异氟醚组只固定不针刺。麻醉暴露：Iso组和EA组由30%氧气和70%氮气混合气体输送异氟醚通过麻醉箱行麻醉暴露,调整麻醉箱内异氟醚浓度至1.2%并维持稳定；Con组同时置于另外相同麻醉箱内,吸入30%氧气和70%氮气混合气体4h。Morris水迷宫测试小鼠学习记忆能力的变化；TUNEL检测海马CA1区细胞凋亡变化；免疫组化检测各组海马CA1区激活型caspase-3、Aβ的表达；Western blot检测海马APP-C83及APP-C99的表达。结果：一、行为学测试 (一)AD小鼠与野生学习及记忆能力比较：各组小鼠的逃避潜伏期随着测试天数的增加逐渐缩短；AD小鼠的逃避潜伏期长于野生小鼠,穿越平台次数及第一象限停留时间百分比低于野生小鼠,差异有统计学意义(P0.05)。 (二)AD小鼠在水迷宫测试中的成绩：定位航行测试中逃避潜伏期比较：T-ConT-EAT-Iso, T-Iso与T-Con之间有显著性差异(P0.05),T-EA与T-Iso间差异有统计学意义(P0.05)。空间探索实验比较：第一象限停留时间百分比T-ConT-EAT-Iso,T-Iso与T-Con差异有统计学意义(P0.05),T-EA与T-Iso差异有统计学意义(P0.05)；AD小鼠60s内穿越平台次数T-ConT-EAT-Iso, T-Iso及T-EA与T-Con差异有统计学意义(P0.05)。 (三)野生小鼠在水迷宫测试中的成绩：定位航行测试中逃避潜伏期比较：W-ConW-EAW-Iso,W-Iso与W-Con之间有显著性差异(P0.05),W-EA与W-Iso间差异有统计学意义(P0.05)。空间探索实验比较：野生组小鼠在第一象限停留时间百分比及60s内穿越平台次数比较：W-ConW-EAW-Iso,三组之间差异无统计学意义(P0.05)。二、激活型caspase-3的表达及TUNEL细胞凋亡情况比较 (一)AD小鼠：AD小鼠海马区caspase-3阳性细胞数目比较：T-ConT-EAT-Iso,T-Iso与T-Con间差异统计学意义(P0.05), T-EA与T-Iso间差异有统计学意义(P0.05)。AD小鼠海马区TUNEL细胞凋亡情况与激活型caspase-3的表达相一致。 (二)野生小鼠：野生小鼠海马区caspase-3阳性细胞数目比较：W-ConW-EAW-Iso,三组之间差异无统计学意义(P0.05)。野生小鼠海马区TUNEL细胞凋亡情况与激活型caspase-3的表达相符。三、APP-C83、APP-C99及Aβ表达情况 (一) Western blot检测APP-C83及APP-C99的表达：APP-C83/GAPDH:T-Iso组APP-C83的表达低于T-Con和W-Con (P0.05); T-EA组APP-C83的表达高于T-Iso组(P0.05). APP-C83/GAPDH:T-Iso组APP-C99的表达高于T-Con和W-Con; T-EA组APP-C99的表达低于T-Iso组(P0.05)。 (二)免疫组化检测Aβ的表达：6月龄AD小鼠海马区已有明显的老年斑形成；T-Iso及T-EA组Aβ表达高于T-Con组(P0.05)；T-EA组A β表达低于T-Iso组,但两组之间差异无统计学意义(P0.05)。结论：一、6月龄AD小鼠的学习记忆能力较野生小鼠已经有明显下降。二、电针可在一定程度上减轻异氟醚诱使的AD小鼠的认知功能损害。三、电针可通过抑制异氟醚引起的AD小鼠海马区激活型caspase-3的过度表达,减少神经元凋亡,减轻异氟醚对AD小鼠造成的神经细胞损害。四、电针可抑制异氟醚导致的APP-C99的过量表达,一定程度上减少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.
【学位授予单位】：广州中医药大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R245

【参考文献】