调理肝脾气机法对更年期大鼠卵巢功能的调控

Effect of regulating qi movement of liver and spleen on ovarian function of climacterical rats

目的:采用从调理肝脾气机入手,间接滋养先天的方法,观察调理肝脾气机法对更年期大鼠卵巢功能的调控作用。方法:①实验于2002-09/11在四川省中药所药理实验室完成。采用SD健康雌性大白鼠62只,其中14～16月龄52只,3月龄10只,动物购进后适应性喂养3天后进行实验。将52只14～16月龄实验大鼠随机分为4组:更年期组(12只)、尼尔雌醇组(12只)、调理肝脾高剂量组(14只)、调理肝脾低剂量组(14只);另10只3月龄正常大鼠设为青年组。②更年期组、青年组灌胃给予蒸馏水10mL/(kg·d);尼尔雌醇组灌胃每5d给予尼尔雌醇犤由上海华联制药有限公司生产,批号:010801,批准文号:沪卫药准字(1995)第012090号犦混悬液0.5mg/kg,调理肝脾气机药物(主要成分为柴胡,由四川省中药研究所提供,批号:020318)。高、低剂量组灌胃给予调理肝脾气机药物溶液6和3g/(kg·d),均连续用药15d。③称取子宫质量,计算子宫湿质量指数(脏器质量/100g体质量)。同时用放射免疫法测定血清促卵泡生成激素、促黄体生成激素、雌二醇水平。采用XSJ-2型光学显微镜观测大鼠卵巢组织态学变化。计数成熟卵泡、生长卵泡的数目、卵巢颗粒细胞层面积、黄体数量。④计量资料差异比较采用单因素方差分析,组间比较采用LSD分析。结果:实验过程中因部分大鼠取样不合格而脱失,观察大鼠子宫湿重指数和血清雌激素水平时尼尔雌醇组脱失1只,进行卵巢组织态学观察时更年期组,青年组,尼尔雌醇组,调理肝脾气机药物高、低剂量组脱失2,2,3,3,3只。①子宫湿重指数:更年期组与青年组接近(P>0.05);尼尔雌醇组和调理肝脾气机药物低剂量组明显高于更年期组(P<0.01,0.05);调理肝脾气机药物高剂量组明显低于尼尔雌醇组(P<0.01)。②青年组大鼠血清促卵泡生成激素水平高于更年期组,但差异不明显(P>0.05),促黄体生成激素、雌二醇水平明显低于更年期组(P<0.05)。调理肝脾气机药物高、低剂量组大鼠血清促卵泡生成激素水平明显高于更年期组(P<0.05,0.01),促黄体生成激素水平明显低于更年期组(P<0.01),雌二醇水平低于更年期组,但差异不明显(P>0.05)。③更年期组大鼠卵巢生长卵泡数量和卵巢颗粒细胞层面积明显少于或小于青年组和调理肝脾气机药物高、低剂量组(P<0.05～0.01)。结论:调理肝脾气机对更年期大鼠血中激素水平影响,不是单纯升高,而是综合性的调节作用。通过改善卵巢衰老,促进卵泡发育、黄体形成,改善体内性激素环境,调整性腺轴功能,使血清性激素水平基本恢复到正常状态。

AIM; To observe effect of regulating qi movement of liver and spleen on regulating and controlling ovarian function of climacterical rats through the method of nourishing innate essence indirectly by regulating qi movement of liver and spleen, METHODS： ① This experiment was Completed in the Pharmacological Laboratory of Sichuan Institute of Traditional Chinese Medicine （TCM） from November to December 2002. Totally 62 health female rats which include 52 rats of fourteen to sixtean months old for first group and 10 rats of three months old for second group. The selected rats were fed for three days in order to adapt to the next experiment. The first group was divided into four groups including 12 rats for climacterical group, 12 rats for nilbstriol group, 14 rats for highdose group and tile rest for low-dose group. The second group was young group, ② Rats in the climacterical group and the young group were perfused with 10 mL/（kg·d） distilled water. At the same time, rats in the nilestriol group were perfused with nilestriol （Shanghai Hualian Pharmacy Company Limited, batch number： 012090, apprevaled number：, hu wei yao zhun zi （1995） 012090] 0.5 mg/kg suspending liquid per five days, regulating qi movement of liver and spleen （main component： Chinese thorowax root; Sichuan Institute of Traditional Chinese Medicine, batch number： 020318）. Rats in high-dose group were perfused with the 6 g/（kg.d） Chinese herb and 3 g/（kg.d） for the low-dose group for 15 days. ③ The quality index of fresh uteruses was calculated as the formula： the weight of rat＇s uterus and ovary/the weight of the rat）. While the contents of folliculin, leuteinizing hormone and estradiol in blood were measured with radio-immunity method. Changes of the ovaries were observed with XSJ-2 optical microscope. And the numbers of mature follicles, growing follicles and corpora lutea were calculated, and also the surface area of granule cells of ovaries was calculated. ④Differences from measurement data were analyzed with single factor mean square method, while inter-block data were compared with LSD method. RESULTS： The unqualified data of some rats were eliminated from the experiment. One rat＇s data were lost in nilestriol group during observing quality index of fresh uteruses and content of estrogen in blood, Two rats＇ data were lost in both climacterical group and young group during observing tectology change of the ovaries. Also three rats＇ data were in the rest groups. ① Quality index of fresh uteruses： The indexes were closed between elimacterical group and young group （P 〉 0.05）. The indexes of nilestriol group and low-dose group were higher than that of climacterical group （P 〈 0.01, 0.05）. And the index of high-dose group was much lower than that of nilestriol group （P 〈 0,01）. ② The content of hemo folliculin in young group was little higher than that in climaeterical group （P 〉 0.05）, However the contents of both hemo leuteinizing hormone and estradiol in young group were much lower than those in climacterical group （P 〈 0.05）. The contents of hemo folliculin in both high-dose group and low-dose group were much higher than those in climacterical group （P 〈 0.05, 0.01）. While the content of hemo leuteinlzing hormone was much lower than that in the climaeterieal group （P 〈 0.01）. And the content of hemo estradiol was little lower than that in the climaetefical group （P 〉 0.05）. ③ The number of growing follicles in climacterical group was much less than that in young group, highdose group and low-dose group （P 〈 0.05-0.01）. And also the surface area of granule cells of ovaries was. CONCLUSION： The regulative effect on hemo hormone of climacterical rats is not only to rise but also to regulate comprehensively through regulating qi movement of liver and spleen. This method can delay the aging of ovary, promote the growing of follicles and the luteal forming, improve the state of hormone and adjust sexual gland axis makes the lever of hemo hormone restored normal state.