目的:探讨中药方剂五加丹方对去卵巢雌性大鼠下丘脑组织匀浆中PLC-β1、IP3、IP3R、CaM及PLC?β1、IP3R1、GnRHmRNA和GnRH、CaM蛋白的影响,试图阐释五加丹方对PLC?β/IP3/CaM通路及绝经综合征的影响.方法:取雌性SD大鼠64只,随机分为空白组(NG)、模型组(MG)、中药对照组(TMC)、西药对照组(WMC)各10只,五加丹方高(WH)、中(WM)、低剂量组(WL)各8只.除NG组外,其余6组均手术摘除大鼠双侧卵巢复制去势模型.模型复制成功后,各组采用灌胃给药1次/日,TMC组及WMC组分别给予相应剂量更年安片混悬液及补佳乐混悬液灌胃,NG组、MG组予等剂量生理盐水灌胃.4周后,处死大鼠,检测脑组织中PLC?β1、IP3、IP3R、CaM的含量;PCR法检测PLC?β、IP3R1、GnRH mRNA表达;Western Blot法检测各组脑组织CaM和GnRH蛋白的表达.结果:与NG组比较,大鼠去势后下丘脑组织PLC?β1、IP3、IP3R、CaM含量明显升高(P<0.01),PLC?β1、IP3R1、GnRHmRNA的表达显著升高(P<0.01),GnRH、CaM蛋白表达较空白组均升高(P<0.01),五加丹方可降低去势模型大鼠下丘脑组织中PLC?β1、IP3、IP3R、CaM水平,并下调PLC?β1、IP3R1、GnRHmRNA的表达.结论:五加丹方下调GnRH的表达,可能是通过作用于GnRH上游调节因子PLC?β1、IP3、IP3R、CaM等而产生,从而起到协调生殖轴功能,改善绝经综合征患者生殖内分泌水平.

Objective:To explore the effect of wujiadan Recipe on the expression of FSH, E2 in serum and KiSS-1, GPR54, GnRH mRNA in hypothalamic arcuate nucleus tissue homogenate of ovariectomized rats, and to further explore the mechanism of wujiadan Recipe on menopausal syndrome.Methods:Sixty-four adult female rats were randomly divided into three groups: blank group, model group, Chinese medicine control group, western medicine control group, and wujiadan high, medium and low dose groups. Except for the blank group, all the other groups were ovariectomized. The rats in the normal group and the model group were given the same dose of saline once a day. One rat in the blank group died during the experiment. After 4 weeks of continuous administration, blood was collected from abdominal aorta and brain stereotaxic localization was performed to preserve the arcuate nucleus of hypothalamus. The levels of FSH and E2 in serum were detected by enzyme-linked immunosorbent assay (ELISA), and the expression of Kiss-1, GPR54 and GnRH mRNA in arcuate nucleus of hypothalamus were detected by PCR.Results:1. Compared with the blank group, FSH level in the model group was significantly higher (P < 0.01), and E2 level was significantly lower (P < 0.01). Compared with the model group, the level of serum E2 in the western medicine control group increased significantly (P. The serum FSH content in each group decreased, especially in the high dose group and the western medicine control group (P < 0.01), and in the Chinese medicine control group, the middle dose group and the low dose group (P < 0.05). Compared with the blank group, the expression of KiSS-1 mRNA, GPR54 mRNA and GnRH mRNA in the model group increased significantly (P < 0.01); compared with the model group, the expression of KiSS-1 mRNA and GPR54 mRNA in the high dose group, middle dose group and Western medicine control group of wujiadan prescription decreased significantly (P < 0.05). Compared with the model group, the expression of GnRH mRNA in the high dose group and the western medicine control group decreased significantly (P < 0.01); the middle dose group of wujiadan decreased significantly (P < 0.05).Conclusion: Wujiadan recipe can improve the serum sex hormone level of castrated rats, and reduce the over expression of KiSS-1, GPR54 and GnRH mRNA in hypothalamic arcuate nucleus of castrated rats, which may be the reproductive endocrine mechanism of wujiadan recipe in the treatment of menopausal syndrome.

从古至今,中医诊病,离不开望、闻、问、切(四诊).问诊虽谓之"工"不及"神、圣"之境界,然而临证确是极为重要的.指出:"诊病不问其始,忧患饮食之失节,起居过度,或伤于毒,不先言此,卒持寸口,何病能中?".尤其是对致病的心理因素,往往是通过"数问其情,以引其意"来发掘.