电针“神门”、“三阴交”对CRH注射失眠模型大鼠HPAA功能的影响研究

发布时间：2018-06-13 10:30

本文选题：失眠 + 下丘脑-垂体-肾上腺轴　；参考：《成都中医药大学》2016年硕士论文

【摘要】：目的：观察电针“神门”、“三阴交”对CRH注射失眠模型大鼠睡眠-觉醒、情绪状态以及HPAA功能的影响,揭示电针治疗失眠的中枢机制和关键性神经递质。方法：将健康清洁级SD大鼠根据体重及旷场实验检测筛选出合格的实验大鼠,并将合格的实验大鼠随机分为空白组、假注射组、CRH模型组、CRH电针组。对CRH模型组、CRH电针组大鼠采取侧脑室注射CRH法建立失眠模型,假注射组只在侧脑室插入注射针头但不给药。侧脑室注射后进行24小时昼夜自发活动及旷场实验检测。检测完成后对CRH电针组进行电针“神门”、“三阴交”治疗,每晚20：00开始治疗,1次/天,连续治疗4天。治疗后对各组大鼠进行24小时昼夜自发活动、旷场实验检测。行为学检测结束后,取股动脉血以及丘脑、脑干,采用酶联免疫法检测各组大鼠外周血浆ACTH、CORT、5-HT和中枢CRH、5-HT含量。结果：1.24小时昼夜自发活动：①经侧脑室注射后：白天活动量：CRH模型组、CRH电针组大鼠24小时昼夜自发活动白天活动量与空白组相比明显增多,均具有统计学意义(P0.05)；CRH模型组、CRH电针组大鼠24小时昼夜自发活动白天活动量与假注射组相比明显增多,均具有统计学意义(P0.05)；夜晚活动量：假注射组大鼠24小时昼夜自发活动夜晚活动量与空白组相比略有减少,具有统计学意义(P0.05)；CRH模型组、CRH电针组大鼠24小时昼夜自发活动夜晚活动量与空白组相比明显减少,均具有统计学意义(P0.05)；CRH模型组、CRH电针组大鼠24小时昼夜自发活动夜晚活动量与假注射组相比明显减少,均具有统计学意义(P0.05)；②治疗后：白天活动量：CRH模型组大鼠白天活动量与空白组相比明显增多,CRH电针组大鼠电针治疗后白天活动量与空白组相比略有增多,均具有统计学意义(P0.05)；CRH模型组大鼠白天活动量与假注射组相比明显增多,CRH电针组大鼠电针治疗后白天活动量与假注射组相比略有增多,均具有统计学意义(P0.05)；CRH电针组大鼠电针治疗后白天活动量与CRH模型组相比明显减少,具有统计学意义(P0.05)；夜晚活动量：CRH模型组大鼠夜晚活动量与空白组相比明显减少,差异具有统计学意义(P0.05)；CRH模型组大鼠夜晚活动量与假注射组相比明显减少,差异具有统计学意义(P0.05)：CRH电针组大鼠电针治疗后夜晚活动量与CRH模型组相比明显增多,差异具有统计学意义(P0.05)。2.旷场实验：①基线：各组旷场实验外周格得分、中央格得分、垂直得分均采用单因素方差分析,差异没有统计学意义,说明基线旷场实验检测指标结果均衡,具有可比较性。②经侧脑室注射后：外周格得分：CRH模型组、CRH电针组外周格得分与空白组相比明显减少,均具有统计学意义(P0.05)；CRH模型组、CRH电针组外周格得分与假注射组相比明显减少,均具有统计学意义(P0.05)；中央格得分：CRH模型组中央格得分与空白组相比增多,CRH电针组中央格得分与空白组相比明显增多,均具有统计学意义(P0.05)；CRH电针组中央格得分与假注射组相比略有增多,具有统计学意义(P0.05)；垂直得分：CRH模型组、CRH电针组垂直得分与空白组相比明显增多,均具有统计学意义(P0.05)：CRH模型组、CRH电针组垂直得分与假注射组相比明显增多,均具有统计学意义(P0.05)：③治疗后：外周格得分：CRH模型组大鼠旷场外周格得分与空白组相比明显减少,CRH电针组大鼠电针治疗后旷场外周格得分与空白组相比略有减少,均具有统计学意义(P0.05)；CRH模型组大鼠旷场外周格得分与假注射组相比明显减少,CRH电针组大鼠电针治疗后旷场外周格得分与假注射组相比略有减少,均具有统计学意义(P0.05)；CRH电针组大鼠电针治疗后外周格得分与CRH模型组相比明显增多,具有统计学意义(P0.05)；中央格得分：CRH模型组大鼠中央格得分与空白组相比明显增多,CRH电针组大鼠电针治疗后中央格得分与空白组相比略有增多,均具有统计学意义(P0.05)；CRH模型组大鼠中央格得分与假注射组相比明显增多,CRH电针组大鼠电针治疗后中央格得分与假注射组相比略有增多,均具有统计学意义(P0.05)；CRH电针组大鼠电针治疗后中央格得分与CRH模型组相比明显减少,具有统计学意义(P0.05)；垂直得分：CRH模型组大鼠旷场垂直得分与空白组相比明显增多,CRH电针组大鼠电针治疗后旷场垂直得分与空白组相比略有增多,均具有统计学意义(P0.05)；CRH模型组大鼠旷场垂直得分与假注射组相比明显增多,CRH电针组大鼠电针治疗后旷场垂直得分与假注射组相比略有增多,均具有统计学意义(P0.05)；CRH电针组大鼠电针治疗后垂直得分与CRH模型组相比明显减少,具有统计学意义(P0.05)：3.血浆神经递质含量：①CRH模型组大鼠外周血浆中5-HT、ACTH、CORT量与空白组、假注射组相比均明显增多,差异均具有统计学意义(P0.05)；②CRH电针组大鼠电针治疗后外周血浆中5-HT、ACTH、CORT量与CRH模型组相比均明显减少,差异均具有统计学意义(P0.05)4.中枢神经递质含量：①CRH模型组大鼠丘脑、脑干神经递质CRH含量与空白组、假注射组相比均明显增多,差异具有统计学意义(P0.05)；CRH电针组大鼠电针治疗后丘脑、脑干神经递质CRH含量与CRH模型组相比明显减少,差异具有统计学意义(P0.05)；②CRH模型组大鼠丘脑、脑干神经递质5-HT含量较空白组、假注射组相比均明显减少,差异均具有统计学意义(P0.05)；CRH电针组大鼠电针治疗后丘脑、脑干神经递质5-HT含量较CRH模型组明显增多,差异具有统计学意义(P0.05)。结论：侧脑室注射CRH可造成失眠模型,失眠模型大鼠下丘脑-垂体-肾上腺轴兴奋性增高；电针“神门”、“三阴交”通过调整下丘脑-垂体-肾上腺轴的过度兴奋治疗失眠,下丘脑室旁核CRH是电针治疗失眠症的关键物质。
[Abstract]:Objective: To observe the effects of "Shen men" and "Sanyinjiao" on sleep - Awakening, emotional state and HPAA function of CRH rats injected with insomnia model, and to reveal the central mechanism and key neurotransmitters of electroacupuncture in the treatment of insomnia. Methods: the healthy and clean SD rats were screened out of the qualified experimental rats according to the test of body weight and open field test. The qualified experimental rats were randomly divided into blank group, false injection group, CRH model group and CRH electroacupuncture group. In the CRH model group, the rats in the CRH electroacupuncture group were injected with the lateral ventricle with CRH to establish the insomnia model. The false injection group only inserted the injection needle in the lateral ventricle, but did not give the drug. The lateral ventricle was injected after 24 hours of spontaneous activity and open field test. After the test was completed, the CRH electroacupuncture group was treated by Electroacupuncture of "Shen gate" and "Three Yin cross" treatment. The treatment was started at 20:00 every night, 1 times a day and 4 days. After the treatment, the spontaneous activities of 24 hours and the open field test were carried out for the rats in each group. After the test of behavior, the blood of the femoral artery and the thalamus, brain stem were taken and the enzyme linked immunosorbent assay was used to detect each group. Rat peripheral plasma ACTH, CORT, 5-HT and central CRH, 5-HT content. Results: 1.24 hours of day and night spontaneous activity: (1) after the lateral ventricle injection: daytime activity: CRH model group, CRH electroacupuncture group 24 hours day and night spontaneous activity in day and night activity increased significantly compared with the blank group, all with statistical significance (P0.05); CRH model group, CRH electroacupuncture. The day and night activity of the rats in the 24 hours was significantly higher than that in the false injection group (P0.05), and the night activity of the rats in the 24 hour day and night decreased slightly compared with the blank group (P0.05), and the CRH model group and the CRH electroacupuncture group had 24 hours of day. The night activity of spontaneous night activity decreased significantly compared with that of the blank group (P0.05). CRH model group, the night activity of spontaneous activity in the 24 hour day and night of rats in the CRH electroacupuncture group was significantly lower than that of the false injection group (P0.05). (2) the daytime activity after treatment: the daytime activity of the CRH model group rats Compared with the blank group, the amount of daytime activity in the CRH electroacupuncture group increased slightly compared with that of the blank group (P0.05), and the amount of daytime activity in the CRH model group was significantly higher than that in the false injection group. The daytime activity of the electroacupuncture group of the CRH electroacupuncture group was slightly more than that of the false injection group. There was statistical significance (P0.05); in the CRH electroacupuncture group, the daytime activity after electroacupuncture was significantly lower than that of the CRH model group, with statistical significance (P0.05); the night activity in the CRH model group decreased significantly compared with the blank group in the CRH model group, and the difference had the statistical significance (P0.05); the activity of night activity in the CRH model group and the rats in the CRH model group were significantly higher than those in the CRH model group. Compared with the false injection group, the difference was statistically significant (P0.05) in the:CRH electroacupuncture group, the activity of electroacupuncture at night was significantly higher than that of the CRH model group. The difference was statistically significant (P0.05).2. open field experiment: (1) baseline: the score of the outer circumference, the score of the central lattice and the vertical score were all single factor. Difference analysis, the difference was not statistically significant, indicating that the results of the baseline field test were balanced and comparable. After the injection of the lateral ventricle, the score of peripheral blood in the CRH model group, the peripheral peripheral score of the CRH electroacupuncture group was significantly lower than that in the blank group, and all had statistical meaning (P0.05); the CRH model group, the peripheral peripheral score of CRH electroacupuncture group were scored and Compared with the false injection group, the score of the central lattice was statistically significant (P0.05). The score of central grid in the CRH model group was more than that in the blank group. The score of the central grid in the CRH electroacupuncture group was significantly higher than that in the blank group. All the scores were statistically significant (P0.05). The score of the central grid in the CRH electroacupuncture group was slightly more than that of the false injection group. Study significance (P0.05); vertical score: CRH model group, the vertical score of CRH electroacupuncture group was significantly increased compared with the blank group, all had statistical significance (P0.05):CRH model group, the vertical score of CRH electroacupuncture group was significantly increased compared with the false injection group (P0.05): after treatment: the peripheral score: CRH model group rats open. Compared with the blank group, the score of the OTC week in the CRH electroacupuncture group was significantly lower than that in the blank group (P0.05). The score of the open field week in the CRH model group was significantly lower than that in the false injection group. The score of the open field week after the electroacupuncture group of the CRH electroacupuncture group was compared with that of the false injection group. Compared with the CRH model group, the score of the rats in the CRH electroacupuncture group was significantly higher than that of the CRH model group (P0.05), and the central grid score of the CRH model group was significantly higher than that in the blank group, and the central grid in the CRH model group was significantly higher than that in the blank group, and the center of the CRH electroacupuncture group was in the center of the electroacupuncture group after the treatment. The score of the grid was a slight increase compared with that in the blank group (P0.05); the score of the central grid in the CRH model group was significantly higher than that in the false injection group. The score of the central grid in the electroacupuncture group of the CRH electroacupuncture group was slightly higher than that in the false injection group, and all had statistical meaning (P0.05), and the central lattice of the electroacupuncture group of the CRH electroacupuncture group after the electroacupuncture treatment. The score was significantly lower than that of the CRH model group (P0.05), and the vertical score of the rats in the CRH model group was significantly higher than that in the blank group. The vertical score of the open field after the electroacupuncture group of the CRH electroacupuncture group was slightly increased compared with the blank group (P0.05), and the open field of the CRH model group was perpendicular to the open field. The scores of the rats in the CRH electroacupuncture group increased slightly compared with those in the false injection group, and there was a statistical significance (P0.05). The vertical score of the electroacupuncture group in the CRH electroacupuncture group was significantly lower than that in the CRH model group, with a statistically significant (P0.05):3. plasma neurotransmitter content: (1) the amount of 5-HT, ACTH, CORT in the peripheral plasma of the CRH model group was significantly increased with the blank group, and the difference was statistically significant (P0.05). (2) the plasma 5-HT, ACTH, CORT in the peripheral plasma of the CRH electroacupuncture group were significantly decreased compared with the CRH model group, and the difference was statistically significant (P0.05) 4. central nerve delivery. Content: (1) CRH model group rats thalamus, brain stem neurotransmitter CRH content and blank group, compared with the false injection group was significantly increased, the difference was statistically significant (P0.05); CRH electroacupuncture group rats after electroacupuncture treatment of thalamus, brainstem neurotransmitter CRH content compared with the CRH model group significantly decreased, the difference was statistically significant (P0.05); (2) CRH model The content of 5-HT in the thalamus and brainstem neurotransmitter in the rat model group was significantly lower than that in the blank group, and the difference was statistically significant (P0.05). The content of 5-HT in the thalamus and brainstem neurotransmitter in the CRH electroacupuncture group was significantly higher than that in the CRH model group, and the difference was statistically significant (P0.05). Conclusion: the lateral ventricle injection of CRH can be made. As a model of insomnia, the hypothalamus pituitary adrenal axis excitability of the rat model of insomnia is increased; Electroacupuncture "Shen gate" and "Three Yin" are used to treat insomnia by adjusting the hyperexcitation of the hypothalamus pituitary adrenal axis. The paraventricular nucleus CRH of the inferior colliculus is the key substance of electroacupuncture in the treatment of insomnia.
【学位授予单位】：成都中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R245.97;R-332

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