精乌胶囊治疗更年期综合征的药效学研究

发布时间：2018-05-31 04:24

本文选题：更年期综合征 + 精乌胶囊　；参考：《北京中医药大学》2014年硕士论文

【摘要】：更年期综合征(Climacteric syndrome, CS)是妇女绝经前后由于性激素波动或减少而引起的一系列以自主神经系统功能紊乱为主,伴有神经心理症状的一组症候群(包括潮红潮热、烦躁易怒、失眠健忘、头晕、头疼、腰疼、骨质疏松等)。卵巢功能衰退引起的雌激素水平降低,以及相伴随的神经内分泌紊乱和植物神经功能紊乱直接导致了更年期综合征的发生。精乌胶囊主要由制何首乌、黄精(制)、女贞子(制)、墨旱莲4味中药组成,具有补肝肾、益精血、壮筋骨之功效,拟用于更年期综合征的治疗。本研究探索了更年期综合征动物模型,建立了更接近更年期综合征临床症状的病-证结合动物模型(去势加肾虚),模拟了更年期综合征的临床表现：血液中下丘脑-垂体-性腺轴的相关激素发生改变,植物神经功能的紊乱等。并在此模型上,观察了精乌胶囊对更年期综合征的改善作用,为该复方的临床应用提供实验依据。 1观察去势大鼠模型各指标的变化及精乌胶囊的干预作用目的观察去势(10周)模型大鼠各指标的变化及精乌胶囊对其的影响。方法健康雌性,育龄未孕SD大鼠90只,随机留取15只为正常对照组,其余大鼠均手术摘除双侧卵巢。自第6-10天,每日对大鼠进行阴道涂片检查,以不出现动情期反应为去势成功。大鼠去势6周后,随机分为：去势模型组、精乌胶囊1.62、0.81、0.405g/kg三个剂量组,坤宝丸组(0.823g/kg)、按照相应剂量灌胃给药,正常对照组及去势模型组给于等量饮用水,连续4周。给药期限结束后,各组动物进行行为学指标观察(敞箱活动、自主活动、对戊巴比妥钠诱导睡眠时间的影响)及直肠温度测定；动物麻醉后取血,测全血粘度；部分血液离心取血清,测定血清孕酮(P)、雌二醇(E2)、黄体生成素(LH)、卵泡刺激素(FSH)、催乳素(PRL)、促性腺激素释放激素(GnRH)含量；各组动物处死取脏器(子宫、肾、肾上腺)称重,计算脏器系数。结果去势(10周)模型组大鼠血清相关激素的变化是：GnRH显著下降(P0.01),P含量降低1/2(因标准差大,未见显著性差异)；子宫系数显著下降(P0.01),肾系数显著下降(P0.05)；直肠温度显著下降(p0.01)；各切率(1s-1、30s-1、70s-1、100s-1、150s-1、200s-1)全血粘度显著增大(P0.01)。其余指标未见明显变化。与去势(10周)模型组比较,精乌胶囊在某些指标上有逆转作用,表现在各剂量大鼠直肠温度回升(P0.05或P0.01),GnRH含量回升。但本次研究中去势(10周)模型大鼠血清E2、FSH、LH、PRL水平的变化与文献报道有差异,初步分析与指标检测时间点有关,文献中多在去势后5-6周。结论由于本实验复制的去势模型未显示更年期综合征典型临床表现,故不能用于评价精乌胶囊的作用,仅作为方法学的探索。 2去势加肾虚类更年期综合征状态模型的建立及精乌胶囊的干预作用 2.1去势加肾虚大鼠类更年期综合征状态的模型的建立目的建立去势加肾虚大鼠模型,以评价该模型的特征。方法健康雌性,育龄未孕SD大鼠50只,随机留取15只作为正常对照组,其余大鼠均手术摘除双侧卵巢。大鼠去势后,自第5-7天,每日对大鼠进行阴道涂片检查,将不出现动情期反应大鼠随机分为单去势组、去势加肾虚组。去势加肾虚组大鼠每日肌注氢化可的松(25mg/kg),连续两周。每周测大鼠体重；于建模完成时,测大鼠尾根、腋下、足底温度；于建模完成时、建模4周、建模6周检测大鼠血清相关激素(E2、FSH、LH、GnRH)水平；于建模完成前1周,建模完成时,建模1周,建模2周,建模3周,建模5周,建模6周测大鼠直肠温度；于建模6周后,依次观察大鼠行为(敞箱活动、自主活动、戊巴比妥钠诱导睡眠)以及脏器(子宫、肾、肾上腺)系数。结果与正常对照组比较,单去势大鼠各指标变化如下：建模完成时,E2显著下降(P0.01),FSH显著升高(P0.05)；建模4周,GnRH显著下降(P0.01)；建模6周,E2显著下降(P0.05),LH显著升高(P0.05),子宫系数、肾系数均显著降低(P0.01),在敞箱中垂直活动次数显著升高(P0.05)；体重快速增加,直肠温度略有下降。去势加肾虚大鼠各指标变化如下：建模完成时,E2、GnRH均显著下降(P0.01或P0.05),FSH显著升高(P0.01),大鼠足底、腋下温度均显著升高(P0.05或P0.01)；建模4周,E2、GnRH显著下降(P0.05或P0.01)；建模6周,E2显著下降(P0.05),GnRH显著降低(P0.01)；子宫系数显著下降(P0.01),大鼠在敞箱中水平活动次数显著下降(P0.05)；体重缓慢显著增加(P0.01),直肠温度显著下降(P0.05)。结论单去势(8周)和去势加肾虚大鼠,均模拟了更年期激素水平紊乱,子宫萎缩以及体温和行为方面的变化。与单去势(8周)比较,去势加肾虚大鼠表现出的类更年期下丘脑-垂体-性腺轴激素水平紊乱及植物神经功能紊乱状态更加典型。 2.2精乌胶囊对去势加肾虚大鼠类更年期综合征状态的干预作用目的观察精乌胶囊对去势加肾虚大鼠类更年期综合征状态的作用,并初步分析其作用机制。方法健康雌性,育龄未孕SD大鼠100只,随机留取15只作为正常对照组,按2.1中方法建立去势加肾虚大鼠模型,并随机分为：模型组、精乌胶囊1.62、0.81、0.405g/kg剂量组,坤宝丸组(0.823g/kg)。按照相应剂量每天灌胃给药,连续8周后,观察各组大鼠血清激素(E2、P、PRL、LH、FSH、GnRH)、脏器(子宫、肾、肾上腺、垂体、下丘脑、海马)系数、行为(敞箱活动、自主活动、高架十字迷宫、戊巴比妥钠诱导睡眠)、直肠温度、血清氧化产物丙二醛(MDA)水平及脑内(下丘脑、皮层)单胺类神经递质(去甲肾上腺素(NE)、多巴胺(DA)、5-羟吲哚乙酸(5-HIAA)、5-羟色胺(5-HT))水平的变化。结果模型大鼠各指标变化如下：血清P、PRL水平显著下降(P0.01)；子宫、垂体系数显著下降(P0.01)；在敞箱中垂直活动次数显著下降(P0.05),自主活动显著下降(P0.05),高架十字迷宫中：总路程显著下降(P0.01)；大鼠直肠温度显著降低(P0.01)；大鼠下丘脑NE水平显著升高(P0.01),皮层5-HIAA水平显著降低(P0.05)。与模型组比较,精乌胶囊各剂量组各指标变化如下：大剂量显著降低血清GnRH水平(P0.05),小剂量显著降低血清P水平(P0.05)；大剂量子宫系数显著下降(P0.05),大、中、小剂量肾系数显著下降(P0.01或P0.05),中剂量肾上腺系数显著下降(P0.05),大、中剂量下丘脑系数显著下降(P0.05)；大剂量显著升高水平活动次数(P0.05),中剂量显著降低水平活动次数(P0.05),中、小剂量显著升高垂直活动次数(P0.01),大剂量显著增加自主活动次数(P0.01)；高架十字迷宫中：大剂量显著增加进开臂区停留时间百分比(P0.05),中剂量显著增加总路程、开臂区停留时间百分比(P0.01或P0.05),小剂量显著增加总路程(P0.01)；各剂量均可显著降低大鼠血清MDA水平(P0.01)；小剂量下丘脑NE显著升高(P0.01),大、小剂量显著降低皮层DA水平(P0.01或P0.05),大、中、小剂量显著降低皮层5-HIAA水平(P0.01),中剂量显著降低5-HT水平(P0.01),大剂量显著降低皮层5-HT(P0.05)结论精乌胶囊对去势加肾虚动物模型的改善作用不是通过促进下丘脑-垂体-性腺轴的分泌,而是侧重于调节中枢单胺类神经递质的失衡,从而改善行为等的异常。 3小结 1疾病动物模型(去势)与病-证结合动物模型(去势加肾虚)相比较,二者均可模拟更年期激素水平紊乱,子宫萎缩、体重以及体温的变化。但前者需掌握成模的时间,不宜过长,后者表现出类更年期下丘脑-垂体-性腺轴激素水平紊乱状态指标、体温改变,植物神经功能紊乱以及肾虚表征等更典型,对评价中药的作用更适用。 2精乌胶囊具有改善去势加肾虚大鼠类更年期综合征状态的作用。主要特点表现在以下几个方面：对下丘脑-垂体-性腺轴血清相关激素无明显促进分泌作用；增加大鼠自主活动次数,消除焦虑情绪；调节中枢单胺类神经递质水平的紊乱。本研究结果为精乌胶囊的临床应用提供了部分实验依据,其作用特点及深入的作用机制有待进一步的研究。
[Abstract]:Climacteric syndrome (CS) is a series of syndrome (including tidal red tide fever, irritability, insomnia and forgetfulness, dizziness, headache, pain, low back pain, osteoporosis, etc.). The lowering of the estrogen level caused by the retreat, as well as the accompanying neuroendocrine disorder and the disturbance of the plant nerve function directly lead to the occurrence of menopause syndrome. The Jingwu capsule is mainly composed of 4 Chinese herbs, Radix Polygoni Multiflori, Huang Jing (system), Ligustrum lucidum (system), and dry lotus. It has the function of supplementing the liver and kidney, benefiting the blood and the strong muscles and bones. It is intended to be used in the climacteric synthesis. The treatment of signs.
This study explored the animal model of climacteric syndrome, established a disease syndrome combined animal model (castration plus kidney deficiency) which was closer to the clinical symptoms of climacteric syndrome, and simulated the clinical manifestations of menopause syndrome: changes in the hypothalamus pituitary - gonadal axis related hormones in the blood and the disorder of plant nerve function. The effects of Jing Wu Capsule on climacteric syndrome were observed, providing experimental basis for the clinical application of the compound.
1 to observe the changes of indexes in castrated rat models and the intervention effect of Jing Wu capsule.
Objective To observe the changes of the indexes of the castrated (10 week) model rats and the effect of Jing Wu Capsule on it. Methods healthy female and 90 SD rats of childbearing age were randomly left to take 15 normal controls. The other rats were removed from the bilateral ovary by operation. From day 6-10, the rats were smeared with vaginal smears every day, with no estrus reaction as castration. After 6 weeks of castration, rats were randomly divided into two groups: castration model group, three dose group of Jing Wu capsule 1.62,0.81,0.405g/kg, Kun Bao pill group (0.823g/kg), gavage according to the corresponding dosage, normal control group and castrated model group were given equal amount of drinking water for 4 weeks. Action, autonomic activity, the influence of pentobarbital sodium induced sleep time induced by sleep time and rectal temperature determination; after anaesthesia, blood was taken and whole blood viscosity was measured; some blood centrifuged serum, serum progesterone (P), estradiol (E2), luteinizing hormone (LH), follicle stimulating hormone (FSH), prolactin (PRL), gonadotropin releasing hormone (GnRH) content, and the content of gonadotropin releasing hormone (GnRH). The changes of serum related hormones in the model group of the castrated (10 weeks) model group were: GnRH significantly decreased (P0.01), P content decreased 1/2 (the standard difference was large, no significant difference); the uterine coefficient decreased significantly (P0.01), the renal coefficient decreased significantly (P0.05); rectal temperature was significantly lower. The whole blood viscosity increased significantly (1s-1,30s-1,70s-1100s-1150s-1200s-1) (P0.01), and the whole blood viscosity increased significantly (P0.01). There was no obvious change in other indexes. Compared with the castrated (10 week) model group, the capsule of Jing Wu had a reversal effect on some indexes, showing the recovery of rectal temperature in each dose (P0.05 or P0.01), and the increase of GnRH content. The changes of serum E2, FSH, LH and PRL levels in the model rats were different from those in the literature. The preliminary analysis was related to the time point of the test, and the literature was more than 5-6 weeks after the castration. Conclusion because the castration model of this experiment did not show the typical clinical manifestation of the climacteric syndrome, it can not be used to evaluate the effect of Jing Wu capsule, only as a recipe. The exploration of law.
2 Establishment of a climacteric syndrome model with castration plus kidney deficiency and intervention of Jing Wu capsule
2.1 establishment of climacteric syndrome model with castration plus kidney deficiency rats
Objective to establish the model of castration and kidney deficiency rats to evaluate the characteristics of the model. Methods 50 healthy female SD rats and 15 rats of childbearing age were randomly selected as the normal control group. The other rats were removed by operation. After the castration, the rats were smeared with vaginal smears on day 5-7, and no estrus reaction was observed in rats. The rats were divided into single castration group, castration plus kidney deficiency group. The rats were injected with hydrocortisone (25mg/kg) daily for two weeks. The rats' body weight was measured every week. The rat tail root, underarm and foot temperature were measured at the completion of modeling. When Yu Jianmo finished, the model was modeled for 4 weeks, and the model of rat serum related hormone (E2, FSH, LH, GnRH) was established for 6 weeks; and the construction of the rat model was built. 1 weeks before the completion of the model, modeling was completed, modeling for 1 weeks, modeling for 2 weeks, modeling for 3 weeks, modeling for 5 weeks, modeling for 6 weeks and measuring rectal temperature in rats. After modeling for 6 weeks, the rats' behavior (open box activity, autonomic activity, pentobarbital induced sleep) and organs (uterus, kidney, adrenal) coefficient were observed in turn. The results were compared with the normal control group, and the single castrated rats were compared with the normal control group. The changes of the index were as follows: when the modeling was completed, the E2 was significantly decreased (P0.01) and the FSH was significantly increased (P0.05); the modeling was 4 weeks, and GnRH decreased significantly (P0.01); the modeling for 6 weeks, E2 significantly decreased (P0.05), LH significantly increased (P0.05), the uterus coefficient, the kidney coefficient significantly decreased (P0.01), the number of vertical movements in the open box increased significantly (P0.05); the weight of the rectum was rapidly increased and rectum increased. There was a slight decrease in temperature. The changes in the parameters of the ovariectomized and Kidney Asthenia Rats were as follows: when the modeling was completed, E2 and GnRH were significantly decreased (P0.01 or P0.05), FSH was significantly increased (P0.01), the underarm temperature of the rats increased significantly (P0.05 or P0.01), and the modeling for 4 weeks, E2 and GnRH decreased significantly (P0.05 or P0.01); modeling for 6 weeks .01); the uterine coefficient decreased significantly (P0.01), the level of horizontal activity in the open box decreased significantly (P0.05), the body weight slowed significantly (P0.01), and the rectal temperature decreased significantly (P0.05). Conclusion the single castration (8 weeks) and the ovariectomized and Kidney Asthenia Rats all simulated the changes in the hormone levels in the more years, the atrophy of the uterus, and the changes in body temperature and behavior. Compared with the single castration (8 weeks), the hypothalamus - hypophysis - gonadotropin level disorder and the state of the autonomic nervous dysfunction were more typical in the ovariectomized and kidney deficiency rats.
2.2 intervention effects of Jing Wu Capsule on climacteric syndrome in ovariectomized rats and kidney deficiency rats
Objective To observe the effect of Jing Wu Capsule on the state of menopause syndrome in castration and kidney deficiency rats and preliminary analysis of its mechanism. Methods 100 healthy female SD rats and 15 rats of childbearing age without pregnancy were randomly selected as normal control group, and the castrated and kidney deficiency rats were established by 2.1 methods and randomly divided into model group and Jing Wu capsule 1.62,0.81 0.405g/kg dose group and Kun Bao pill group (0.823g/kg). The serum hormone (E2, P, PRL, LH, FSH, GnRH) and the coefficient of the organs (uterus, kidney, adrenal, hypophysis, hypothalamus, hippocampus) of rats were observed for 8 weeks after the corresponding dose of the stomach, and the rectal temperature was (open box activity, elevated cross maze, pentobarbital induced sleep), and rectal temperature. The level of serum oxidation product malondialdehyde (MDA) and the level of monoamine neurotransmitter (norepinephrine (NE), dopamine (DA), 5- hydroxyindolo acetic acid (5-HIAA), 5- hydroxytryptamine (5-HT)) in the brain (hypothalamus, cortex). The results of the model rats were as follows: the serum P, the PRL level decreased significantly (P0.01), the uterus, the pituitary coefficient was significantly lower. Drop (P0.01); the number of vertical activities in the open box decreased significantly (P0.05), autonomous activity decreased significantly (P0.05), in the elevated cross maze, the total path decreased significantly (P0.01), the rectal temperature in rats decreased significantly (P0.01), the level of NE in the hypothalamus of rats was significantly increased (P0.01), and the level of cortical 5-HIAA decreased significantly (P0.05). The changes of the indexes in each dose group were as follows: the large dose significantly reduced the serum GnRH level (P0.05), the small dose significantly decreased the serum P level (P0.05), the large dose of the uterus coefficient decreased significantly (P0.05), the large, medium and small dose renal coefficient decreased significantly (P0.01 or P0.05), the medium dose adrenal coefficient decreased significantly (P0.05), and the medium dose hypothalamic coefficient was significant. Decrease (P0.05); large dose significantly increased level of activity (P0.05), middle dose significantly reduced the frequency of horizontal activity (P0.05), small dose significantly increased the number of vertical activity (P0.01), large dose significantly increased the number of autonomous activities (P0.01), high dose of the cross maze: large dose significantly increased the percentage of stay in the open arm area (P0.05). The median dose increased significantly, the percentage of staying time in the open arm area (P0.01 or P0.05), the small dose significantly increased the total road distance (P0.01), and the serum MDA level (P0.01) was significantly reduced in each dose, and the low dose of NE in the hypothalamus increased significantly (P0.01), and the small dose significantly decreased the level of DA (P0.01 or P0.05) in the cortex (P0.01 or P0.05), and the small dose decreased significantly. Cortical 5-HIAA level (P0.01), middle dose significantly reduced 5-HT level (P0.01), large dose significantly decreased cortical 5-HT (P0.05). Conclusion the effect of Jing Wu Capsule on castration and kidney deficiency animal model is not by promoting the secretion of hypothalamus pituitary gonadal axis, but focusing on regulating the imbalance of the central monoamine neurotransmitters, thus improving the behavior and so on. Abnormality.
3 Summary
1 the animal model of disease (castration) compared with the animal model of disease and syndrome (castration plus kidney deficiency), the two can simulate changes in hormone level disorder, uterine atrophy, body weight and body temperature in menopause. However, the former needs to master the time of mold formation and should not be too long, the latter shows the index of the level disorder of hypothalamus pituitary gonadal hormone levels in the more years. Changes in body temperature, autonomic nervous function and kidney deficiency are more typical, and are more suitable for evaluating the role of traditional Chinese medicine.
The 2 Jing Wu capsule has the effect of improving the state of menopause syndrome of rats with castration and kidney deficiency. The main characteristics are as follows: there is no obvious promotion of the secretion of serum related hormones in the hypothalamus pituitary gonadal axis, increasing the frequency of autonomic activity and eliminating anxiety, and regulating the turbulence of the level of monoamine neurotransmitters in the central nervous system. The results of this study provide some experimental evidence for clinical application of Jing Wu capsule, and its action characteristics and in-depth mechanism need to be further studied.
【学位授予单位】：北京中医药大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R271.9

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