益肾续经汤延缓绝经过渡期大鼠卵巢功能衰老的实验研究
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摘要
研究目的探讨女性绝经过渡期早期的衰老机理,在前期临床研究的基础上,通过建立绝经过渡期早期动物模型,从内分泌调控及Caspase依赖型卵巢凋亡通路两个方面,探讨益肾续经汤在绝经过渡期早期延缓卵巢衰老进程的作用机制。
理论研究绝经过渡期早期标志女性开始衰老,肝血不足是女性衰老早期的典型特点；衰老速度与肾气的强弱密切相关；瘀血是衰老过程的必然产物,血瘀是加速衰老的重要因素。黎烈荣教授针对绝经过渡期早期女性特殊的生理病理基础,兼顾开始虚损的肝肾二脏,养肝血、补肾精的同时,不忘通血脉,以免瘀血阻滞脉络而加速衰老进程,体现了“既病防变”的治未病思想。从女性的最早期衰老阶段着手,在治疗绝经过渡期早期月经不调的同时,中药干预可延缓女性衰老进程,以期推迟绝经年龄,减少绝经过渡期晚期围绝经期综合征的发生,这正是“未病先防”的具体体现。
实验研究
研究方法：本实验以绝经过渡期(Menopausal Transition, MT) wistar雌性大鼠为研究对象,选取阴道脱落细胞涂片表现为动情间期延长、动情周期紊乱者纳入实验。以大鼠月龄区分MT早期(10月龄)和MT晚期(11-12月龄)。选取符合MT早期标准的大鼠24只,随机分为MT早期模型组、MT早期治疗组和西药对照组,每组8只；选取符合MT晚期标准的大鼠16只,随机分为MT晚期模型组、MT晚期治疗组,每组8只；另选取4-6月龄大鼠8只,作为青年对照组(青年组)。MT早期模型组、MT晚期模型组、青年组分别给予生理盐水；MT早期治疗组和MT晚期治疗组给予益肾续经汤；西药组予以去氧孕烯炔雌醇水溶液灌胃。各组每日均按1ml/100g灌胃,连续灌胃15天,末次给药24小时后,腹主动脉取血,分离血清,酶联免疫吸附法检测抑制素B (INHB)、促卵泡素(FSH)、雌二醇(E2)水平；摘取卵巢,以电子天平称取卵巢湿重,将脏器湿重换算成脏器指数。石蜡包埋,常规切片4μm,左侧卵巢切片HE染色用于光镜下观察卵巢形态学表现,免疫组化SP法测定各组大鼠右侧卵巢Fas、Cytc、Caspase-12、Caspase-3的表达。
实验结果：
1、MT早期模型组INHB水平低于青年组(P<0.01),MT晚期模型组低于MT早期模型组(P<0.01)。MT早期治疗组INHB水平显著高于MT早期模型组(P<0.01)。MT晚期治疗组INHB水平与MT晚期模型组INHB水平差异无显著意义(P>0.05)。西药组与MT早期模型组INHB水平差异无显著性(P>0.05)。
2、MT早期模型组FSH水平高于青年组(P<0.01)；MT晚期模型组高于MT早期模型组(P<0.01)。MT早期治疗组FSH水平接近青年组(P>0.05)。西药组FSH水平低于MT早期模型组(P<0.01),但高于青年组(P<0.05)。MT晚期治疗组与MT晚期模型组FSH差异无显著性(P>0.05)
3、MT早期模型组E2水平明显高于青年组(P<0.01),MT晚期模型组显著低于青年组(P<0.01)。使用益肾续经汤后,MT早期治疗组E2水平较MT早期模型组明显降低(P<0.01),MT晚期治疗组高于MT晚期模型组(P<0.01)。
4、青年组卵巢指数高于MT早期模型组(P<0.01),MT早期模型组高于MT晚期模型组(P<0.01)。MT早期治疗组卵巢指数显著高于MT早期模型组(P<0.05)。MT晚期治疗组卵巢指数虽略高于MT晚期模型组,但差异无显著意义(P>0.05)。西药组与MT早期模型组差异无显著性(P>0.05)。
5、光镜下可见,MT早期治疗组和青年组大鼠卵巢内有较多数目的初级卵泡、次级卵泡、窦状卵泡等各级生长发育卵泡,黄体、白体等都可见,卵泡形状规则,体积较大。MT早期模型组和西药组大鼠卵巢皮质内卵泡数目较少,卵泡形状不规则。MT晚期模型组和MT晚期治疗组大鼠卵巢内生长卵泡明显减少,卵泡结构紊乱。
6、MT晚期模型组卵巢Fas表达水平显著高于MT早期模型组(P<0.01)；MT早期模型组明显高于青年组(P<0.01)。使用益肾续经汤后,MT早期治疗组较MT早期模型组卵巢Fas表达水平显著降低(P<0.01),MT晚期治疗组与MT晚期模型组差异无显著性(P>0.05)。西药组卵巢Fas表达水平与MT早期模型组差异无显著意义(P>0.05)。
7、MT晚期模型组卵巢Caspase-12表达水平显著高于MT早期模型组(P<0.05),MT早期模型组明显高于青年组(P<0.01)。MT早期治疗组卵巢Caspase-12表达水平明显低于MT早期模型组(P<0.05),与青年组水平接近(P>0.05)。MT晚期治疗组亦可降低卵巢Caspase-12表达水平(P<0.05)。西药组与MT早期模型组差异无显著意义(P>0.05)。
8、青年组卵巢CytC、Caspase-3表达最低,MT早期模型组高于青年组(P<0.01、P<0.01),MT晚期模型组高于MT早期模型组(P<0.01、P<0.01)。MT早期治疗组卵巢CytC、Caspase-3表达均显著低于MT早期模型组(P<0.01、P<0.05),但高于青年组(P<0.01、P<0.01)。MT晚期治疗组卵巢CytC、Caspase-3表达与MT早期模型组无明显差异(P>0.05、P>0.05)。西药组卵巢CytC、Caspase-3表达低于MT早期模型组,差异无显著性(P>0.05)。
9、未给药组全部大鼠卵巢Caspase-3表达与其他凋亡指标的两因素相关分析显示,Caspase-3与Fas(r=0.453,P<0.001)、Caspase-12 (r=0.798, P<0.001)和CytC (r=0.935, P<0.001)均呈显著正相关。
结论：导师黎烈荣教授提出在绝经过渡期早期以养肝血、补肾精、通血脉立论,拟定益肾续经汤治疗绝经过渡期早期月经不调,延缓女性衰老进程。本课题在前期临床研究疗效确切的基础上,首次建立绝经过渡期早期模型,为研究者未来从实验角度研究绝经过渡期提供了研究基础。实验研究表明,在MT早期使用益肾续经汤,能够升高INHB水平,增强INHB对FSH的负反馈调节,从而降低FSH水平,可以调节卵泡发育和闭锁交替引起的雌激素波动至正常状态。益肾续经汤通过调整紊乱的激素水平,改善激素效应,使激素作用的靶器官蛋白质合成增加,降低卵巢重量下降速度,对生殖轴具有正向调节作用。益肾续经汤能通过降低三条经典凋亡途径的启动凋亡因子Fas、CytC、Caspase-12的表达,从多靶点降低凋亡启始酶的活化或释放,抑制三条Caspase依赖型链式蛋白酶级联反应,继而降低卵巢Caspase-3蛋白的表达,阻止卵巢Caspase依赖型凋亡通路,延缓卵巢老化进程。
Speciality:Gynecology of Traditional Chinese Medicine Author:Yuxinhui
Tutor:LiLieRong
Objects:Probing the Mechanism of aging in early stage of menopausal transition period, and Through establishing the animal model, investigating the mechanism of Yishenxujing decoction in delaying aging of ovary from regulating endocrine secretion and relectting ovarion apoptosis pathway of caspase-depended typle.
Theory probing Deficiency of hepatic blood is the typical feature of aging in early stage of menopausal transition period. The speech of aging is closed with kidney. Blood stasis is products of aging, and blood stasis is one of the most important factors for accelerating speed of aging. Professor li lie rong aims directly at the special physiological functions and pathology of the menopausal transition, giving consideration to Liver and kidney. She suggests delaying the speech of female aging by use of traditional Chinese drug in early stage of menopausal transition period, delay the menopause age and reduce taking place of the menopausal syndromes, this is ref lectting for the theory of prevention and cure redisease. Meanwhile, promoting blood flow to avoide the stagnated blood blocking veins and arteries, which reflects the theory of preventting development for the disease.
Experiment study
Methods:The experiment chose MT female rats as research object, made experiment with vagina cell fitting the model standard. We distinguished the rast as MT initial stage group and MT advanced stage group depend on the age of rats. we distinguished the rats as MT initial stage model group、MT initial stage treatment group and western medicine group from 24 rats which accord with MT initial stage standard. Then we distinguished the rats as MT advanced stage model group and MT advanced stagetreatment group from 16 rats which accord withMT advanced stage standard. Then chose 8 young rats as the young control group. The rats of MT initial stage model group、MT advansed stage model group and the young control group were gavaged with isotonic Na chloride. The rats of MT initial stage treatment group and MT advanced stagetreatment group were gavaged by gastric perfusion of yishenxujing decoction. The Western medicine group received Mafulong by gastric perfusion. After lavaged for 15 days, drawed blood from abdominal aorta, separated the blood serum. The blood sample were tested to detect serum INHB、FSH、E2. The ovaries were treated by HE and Immune histoehemistry method to observed the morphologe of overies、expression of overy Fas、Cytc、Caspase-12 and Caspase-3.
Results:
1. INHB level of young group is higher than MT initial stage model group (P<0.01), and INHB level of MT initial stage model group is higher than MT advanced stage model group (P<0.01). INHB level of MT initial stage treatment group is higher than MT initial model group (P<0.01). INHB level of MT advanced stage treatment group hasn't obvious difference with MT advanced stage group (P<0.05).The western medicine group is approach with MT initial stage model group (P>0.05).
2. The FSH level of MT initial stage model group is higher than young group (P<0.01), and FSH level of MT advanced stage model group is higher than MT initial stage model group (P<0.01).The young group is approach with MT initial stage model group (P>0.05). FSH level of western medicine group is lower than MT initial stage model group (P<0.01), But is higher than the young group (P<0.05).FSH level of MT advanced stage treatment group is approach with MT advanced stage model group (P>0.05).
3. The E2 level of MT initial stage model group is obviously higher than young group (P<0.01).The E2 level of MT advanced stage model group is obviously lower than young group (P<0.01). After using the yishenxujing decoction, E2 level of MT initial stage treatment group is lower than MT initial stage model group (P<0.01), E2 level of MT advanced stage treatment group is higher than MT advanced stage model group (P<0.01).
4. The ovarian index of the young group is higher than MT initial stage model group (P<0.01).The ovarian index of MT initial stage model group is higher than MT advanced stage model group (P<0.01), MT initial stage treatment group is higher than MT advanced stage treatment group (P<0.05), MT advanced stage treatment group is a little higher than MT initial stage model group, but there is not statistical significance (P>0.05). The ovarian index of western medicine group is approach with MT initial stage model group (P>0.05).
5.From the light microscope, we can see there is more primary follicle、secondary follicle、sinusoid follicle in overies of MT initial stage group, and the follicles is regular and big. In ovary cortex of MT initial stage model group and western medicine group rats, there are less follicles, follicles'morphous are irregular. Follicles in ovaries of MT advanced stage treatment group and MT advanced stage model group rats are small and irregular.
6. Fas expression in ovaries of MT advanced stage model group is higher than MT initial stage model group (P<0.05). And the ovaries Fas expression of young group is lower than MT initial stage model menopausal transition group(P<0.01). After using the yishenxujing decoction, Fas expression in ovaries of MT initial stage treatment group is lower than MT initial stage model group (P<0.05). MT advanced stage treatment group hasn't statistical significance with MT initial stage model group (P>0.05). The ovaries Fas expression of western medicine group is approach with MT initial stage model group (P>0.05)
7. Caspase-12 expression in ovaries of MT advanced stage model group is higher than MT initial stage model group (P<0.05). And the ovaries Caspase-12 expression of young group is lower than MT initial stage model group (P<0.01). Caspase-12 expression of MT initial stage treatment group is lower than MT initial stage model group(P<0.05), and apporoach with young group (P>0.05).Expression in ovaries of MT advanced stage expression is lower than MT advanced stage model group(P<0.05), west medicing group is approach with MT initial stage model group (P>0.05).
8. Compared with other groups, CytC、Caspase-3 expression in ovaries of young group is the lowest. And the overies CytC、Caspase-3 expression of MT advanced stage model group is higher than MT initial stage model group (P<0.01、P<0.01).CytC、Caspase-3 expression in overies of MT initial stagetreatment group is lower than MT initial stage model group (P<0.01、P<0.05), but is higher than young group (P<0.01、P<0.01). CytC、Caspase-3 expression of MT advanced stage model group is a little lower than MT initial stage model group (P>0.05、P>0.05). The western medicine group is approach with MT initial stage model group (P>0.05).
Conclusion:The topic erect the initial stage model of the menopausal transition. It makes the foundation for study the menopausal transition from experimentation. My teacher Lilierong makes the principle of giving consideration to Liver and kidney and promoting blood flow, drows up Yisenxujing decoction, which can delay ovarian apoplexies when heating irregular menstruation in early stage of menopausal transition. Experiment invest indicated, yisenxujing decoction could adjust the ovarian follicle development and adjust the estrogenic hormone fluctuation. Yisenxujing decoction could adjust the estrogenic hormone level and amendment the estrogenic hormone effect, improved the protein synthesis. Yisenxujing decoction has the forward direction action for the reproduction genealogy. Yisenxujing decoction could depress expression of Fas、CytC and Caspase-12, depress the activation and dilivery of ovary's apoptosis pathway of caspase depended typle, so that depress expression of Caspase-3 gene, delaying the proceeding of ovarian aging.
理论研究绝经过渡期早期标志女性开始衰老,肝血不足是女性衰老早期的典型特点；衰老速度与肾气的强弱密切相关；瘀血是衰老过程的必然产物,血瘀是加速衰老的重要因素。黎烈荣教授针对绝经过渡期早期女性特殊的生理病理基础,兼顾开始虚损的肝肾二脏,养肝血、补肾精的同时,不忘通血脉,以免瘀血阻滞脉络而加速衰老进程,体现了“既病防变”的治未病思想。从女性的最早期衰老阶段着手,在治疗绝经过渡期早期月经不调的同时,中药干预可延缓女性衰老进程,以期推迟绝经年龄,减少绝经过渡期晚期围绝经期综合征的发生,这正是“未病先防”的具体体现。
实验研究
研究方法：本实验以绝经过渡期(Menopausal Transition, MT) wistar雌性大鼠为研究对象,选取阴道脱落细胞涂片表现为动情间期延长、动情周期紊乱者纳入实验。以大鼠月龄区分MT早期(10月龄)和MT晚期(11-12月龄)。选取符合MT早期标准的大鼠24只,随机分为MT早期模型组、MT早期治疗组和西药对照组,每组8只；选取符合MT晚期标准的大鼠16只,随机分为MT晚期模型组、MT晚期治疗组,每组8只；另选取4-6月龄大鼠8只,作为青年对照组(青年组)。MT早期模型组、MT晚期模型组、青年组分别给予生理盐水；MT早期治疗组和MT晚期治疗组给予益肾续经汤；西药组予以去氧孕烯炔雌醇水溶液灌胃。各组每日均按1ml/100g灌胃,连续灌胃15天,末次给药24小时后,腹主动脉取血,分离血清,酶联免疫吸附法检测抑制素B (INHB)、促卵泡素(FSH)、雌二醇(E2)水平；摘取卵巢,以电子天平称取卵巢湿重,将脏器湿重换算成脏器指数。石蜡包埋,常规切片4μm,左侧卵巢切片HE染色用于光镜下观察卵巢形态学表现,免疫组化SP法测定各组大鼠右侧卵巢Fas、Cytc、Caspase-12、Caspase-3的表达。
实验结果：
1、MT早期模型组INHB水平低于青年组(P<0.01),MT晚期模型组低于MT早期模型组(P<0.01)。MT早期治疗组INHB水平显著高于MT早期模型组(P<0.01)。MT晚期治疗组INHB水平与MT晚期模型组INHB水平差异无显著意义(P>0.05)。西药组与MT早期模型组INHB水平差异无显著性(P>0.05)。
2、MT早期模型组FSH水平高于青年组(P<0.01)；MT晚期模型组高于MT早期模型组(P<0.01)。MT早期治疗组FSH水平接近青年组(P>0.05)。西药组FSH水平低于MT早期模型组(P<0.01),但高于青年组(P<0.05)。MT晚期治疗组与MT晚期模型组FSH差异无显著性(P>0.05)
3、MT早期模型组E2水平明显高于青年组(P<0.01),MT晚期模型组显著低于青年组(P<0.01)。使用益肾续经汤后,MT早期治疗组E2水平较MT早期模型组明显降低(P<0.01),MT晚期治疗组高于MT晚期模型组(P<0.01)。
4、青年组卵巢指数高于MT早期模型组(P<0.01),MT早期模型组高于MT晚期模型组(P<0.01)。MT早期治疗组卵巢指数显著高于MT早期模型组(P<0.05)。MT晚期治疗组卵巢指数虽略高于MT晚期模型组,但差异无显著意义(P>0.05)。西药组与MT早期模型组差异无显著性(P>0.05)。
5、光镜下可见,MT早期治疗组和青年组大鼠卵巢内有较多数目的初级卵泡、次级卵泡、窦状卵泡等各级生长发育卵泡,黄体、白体等都可见,卵泡形状规则,体积较大。MT早期模型组和西药组大鼠卵巢皮质内卵泡数目较少,卵泡形状不规则。MT晚期模型组和MT晚期治疗组大鼠卵巢内生长卵泡明显减少,卵泡结构紊乱。
6、MT晚期模型组卵巢Fas表达水平显著高于MT早期模型组(P<0.01)；MT早期模型组明显高于青年组(P<0.01)。使用益肾续经汤后,MT早期治疗组较MT早期模型组卵巢Fas表达水平显著降低(P<0.01),MT晚期治疗组与MT晚期模型组差异无显著性(P>0.05)。西药组卵巢Fas表达水平与MT早期模型组差异无显著意义(P>0.05)。
7、MT晚期模型组卵巢Caspase-12表达水平显著高于MT早期模型组(P<0.05),MT早期模型组明显高于青年组(P<0.01)。MT早期治疗组卵巢Caspase-12表达水平明显低于MT早期模型组(P<0.05),与青年组水平接近(P>0.05)。MT晚期治疗组亦可降低卵巢Caspase-12表达水平(P<0.05)。西药组与MT早期模型组差异无显著意义(P>0.05)。
8、青年组卵巢CytC、Caspase-3表达最低,MT早期模型组高于青年组(P<0.01、P<0.01),MT晚期模型组高于MT早期模型组(P<0.01、P<0.01)。MT早期治疗组卵巢CytC、Caspase-3表达均显著低于MT早期模型组(P<0.01、P<0.05),但高于青年组(P<0.01、P<0.01)。MT晚期治疗组卵巢CytC、Caspase-3表达与MT早期模型组无明显差异(P>0.05、P>0.05)。西药组卵巢CytC、Caspase-3表达低于MT早期模型组,差异无显著性(P>0.05)。
9、未给药组全部大鼠卵巢Caspase-3表达与其他凋亡指标的两因素相关分析显示,Caspase-3与Fas(r=0.453,P<0.001)、Caspase-12 (r=0.798, P<0.001)和CytC (r=0.935, P<0.001)均呈显著正相关。
结论：导师黎烈荣教授提出在绝经过渡期早期以养肝血、补肾精、通血脉立论,拟定益肾续经汤治疗绝经过渡期早期月经不调,延缓女性衰老进程。本课题在前期临床研究疗效确切的基础上,首次建立绝经过渡期早期模型,为研究者未来从实验角度研究绝经过渡期提供了研究基础。实验研究表明,在MT早期使用益肾续经汤,能够升高INHB水平,增强INHB对FSH的负反馈调节,从而降低FSH水平,可以调节卵泡发育和闭锁交替引起的雌激素波动至正常状态。益肾续经汤通过调整紊乱的激素水平,改善激素效应,使激素作用的靶器官蛋白质合成增加,降低卵巢重量下降速度,对生殖轴具有正向调节作用。益肾续经汤能通过降低三条经典凋亡途径的启动凋亡因子Fas、CytC、Caspase-12的表达,从多靶点降低凋亡启始酶的活化或释放,抑制三条Caspase依赖型链式蛋白酶级联反应,继而降低卵巢Caspase-3蛋白的表达,阻止卵巢Caspase依赖型凋亡通路,延缓卵巢老化进程。
Speciality:Gynecology of Traditional Chinese Medicine Author:Yuxinhui
Tutor:LiLieRong
Objects:Probing the Mechanism of aging in early stage of menopausal transition period, and Through establishing the animal model, investigating the mechanism of Yishenxujing decoction in delaying aging of ovary from regulating endocrine secretion and relectting ovarion apoptosis pathway of caspase-depended typle.
Theory probing Deficiency of hepatic blood is the typical feature of aging in early stage of menopausal transition period. The speech of aging is closed with kidney. Blood stasis is products of aging, and blood stasis is one of the most important factors for accelerating speed of aging. Professor li lie rong aims directly at the special physiological functions and pathology of the menopausal transition, giving consideration to Liver and kidney. She suggests delaying the speech of female aging by use of traditional Chinese drug in early stage of menopausal transition period, delay the menopause age and reduce taking place of the menopausal syndromes, this is ref lectting for the theory of prevention and cure redisease. Meanwhile, promoting blood flow to avoide the stagnated blood blocking veins and arteries, which reflects the theory of preventting development for the disease.
Experiment study
Methods:The experiment chose MT female rats as research object, made experiment with vagina cell fitting the model standard. We distinguished the rast as MT initial stage group and MT advanced stage group depend on the age of rats. we distinguished the rats as MT initial stage model group、MT initial stage treatment group and western medicine group from 24 rats which accord with MT initial stage standard. Then we distinguished the rats as MT advanced stage model group and MT advanced stagetreatment group from 16 rats which accord withMT advanced stage standard. Then chose 8 young rats as the young control group. The rats of MT initial stage model group、MT advansed stage model group and the young control group were gavaged with isotonic Na chloride. The rats of MT initial stage treatment group and MT advanced stagetreatment group were gavaged by gastric perfusion of yishenxujing decoction. The Western medicine group received Mafulong by gastric perfusion. After lavaged for 15 days, drawed blood from abdominal aorta, separated the blood serum. The blood sample were tested to detect serum INHB、FSH、E2. The ovaries were treated by HE and Immune histoehemistry method to observed the morphologe of overies、expression of overy Fas、Cytc、Caspase-12 and Caspase-3.
Results:
1. INHB level of young group is higher than MT initial stage model group (P<0.01), and INHB level of MT initial stage model group is higher than MT advanced stage model group (P<0.01). INHB level of MT initial stage treatment group is higher than MT initial model group (P<0.01). INHB level of MT advanced stage treatment group hasn't obvious difference with MT advanced stage group (P<0.05).The western medicine group is approach with MT initial stage model group (P>0.05).
2. The FSH level of MT initial stage model group is higher than young group (P<0.01), and FSH level of MT advanced stage model group is higher than MT initial stage model group (P<0.01).The young group is approach with MT initial stage model group (P>0.05). FSH level of western medicine group is lower than MT initial stage model group (P<0.01), But is higher than the young group (P<0.05).FSH level of MT advanced stage treatment group is approach with MT advanced stage model group (P>0.05).
3. The E2 level of MT initial stage model group is obviously higher than young group (P<0.01).The E2 level of MT advanced stage model group is obviously lower than young group (P<0.01). After using the yishenxujing decoction, E2 level of MT initial stage treatment group is lower than MT initial stage model group (P<0.01), E2 level of MT advanced stage treatment group is higher than MT advanced stage model group (P<0.01).
4. The ovarian index of the young group is higher than MT initial stage model group (P<0.01).The ovarian index of MT initial stage model group is higher than MT advanced stage model group (P<0.01), MT initial stage treatment group is higher than MT advanced stage treatment group (P<0.05), MT advanced stage treatment group is a little higher than MT initial stage model group, but there is not statistical significance (P>0.05). The ovarian index of western medicine group is approach with MT initial stage model group (P>0.05).
5.From the light microscope, we can see there is more primary follicle、secondary follicle、sinusoid follicle in overies of MT initial stage group, and the follicles is regular and big. In ovary cortex of MT initial stage model group and western medicine group rats, there are less follicles, follicles'morphous are irregular. Follicles in ovaries of MT advanced stage treatment group and MT advanced stage model group rats are small and irregular.
6. Fas expression in ovaries of MT advanced stage model group is higher than MT initial stage model group (P<0.05). And the ovaries Fas expression of young group is lower than MT initial stage model menopausal transition group(P<0.01). After using the yishenxujing decoction, Fas expression in ovaries of MT initial stage treatment group is lower than MT initial stage model group (P<0.05). MT advanced stage treatment group hasn't statistical significance with MT initial stage model group (P>0.05). The ovaries Fas expression of western medicine group is approach with MT initial stage model group (P>0.05)
7. Caspase-12 expression in ovaries of MT advanced stage model group is higher than MT initial stage model group (P<0.05). And the ovaries Caspase-12 expression of young group is lower than MT initial stage model group (P<0.01). Caspase-12 expression of MT initial stage treatment group is lower than MT initial stage model group(P<0.05), and apporoach with young group (P>0.05).Expression in ovaries of MT advanced stage expression is lower than MT advanced stage model group(P<0.05), west medicing group is approach with MT initial stage model group (P>0.05).
8. Compared with other groups, CytC、Caspase-3 expression in ovaries of young group is the lowest. And the overies CytC、Caspase-3 expression of MT advanced stage model group is higher than MT initial stage model group (P<0.01、P<0.01).CytC、Caspase-3 expression in overies of MT initial stagetreatment group is lower than MT initial stage model group (P<0.01、P<0.05), but is higher than young group (P<0.01、P<0.01). CytC、Caspase-3 expression of MT advanced stage model group is a little lower than MT initial stage model group (P>0.05、P>0.05). The western medicine group is approach with MT initial stage model group (P>0.05).
Conclusion:The topic erect the initial stage model of the menopausal transition. It makes the foundation for study the menopausal transition from experimentation. My teacher Lilierong makes the principle of giving consideration to Liver and kidney and promoting blood flow, drows up Yisenxujing decoction, which can delay ovarian apoplexies when heating irregular menstruation in early stage of menopausal transition. Experiment invest indicated, yisenxujing decoction could adjust the ovarian follicle development and adjust the estrogenic hormone fluctuation. Yisenxujing decoction could adjust the estrogenic hormone level and amendment the estrogenic hormone effect, improved the protein synthesis. Yisenxujing decoction has the forward direction action for the reproduction genealogy. Yisenxujing decoction could depress expression of Fas、CytC and Caspase-12, depress the activation and dilivery of ovary's apoptosis pathway of caspase depended typle, so that depress expression of Caspase-3 gene, delaying the proceeding of ovarian aging.
引文
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