| Objectives1. To Analysis the quality of life and its influential factors in patients with deficiency of kidney and blood stasis syndrome of liver stagnation and spleen deficiency type DOR, to provide scientific basis for the deficiency of kidney and blood stasis syndrome of liver stagnation and spleen deficiency DOR prevention and health care, and to provide new ideas and methods for the treatment of kidney deficiency and blood stasis of liver stagnation and spleen deficiency DOR.2. To test success of CTX induced ovarian damage in rat model.3. To observe the effect of Guishen Pill on ovarian function of DOR rats induced by CTX.Methods1. Select50patients to the gynecology clinic, in accordance with the inclusion criteria as the observation group, the other50healthy women as the healthy control group. Women in two groups were using the unified registration information table, and the "health measurement scale SF-36".2. Preparation for test(1) Female rats were intraperitoneally injected with CTX at the dose of75mg/kg, and to test success of CTX induced ovarian damage in rat model.(2)48SD female rats were randomly divided into blank control group, model group, Chinese medicine group. Estrous cycle of rats were observed in each group every day for4weeks after medicine was used.4rats in each group were killed every week, the weight of the ovary, ovarian tissue morphological were observed, follicle number were counted, serum E2、FSH were measured.3. To observe the effect of Guishen Pill on ovarian function of DOR rats model induced by CTX.Results 1. Analysis of the influencing factors and life quality of patients with deficiency of kidney and blood stasis syndrome of liver stagnation and spleen deficiency type DOR.(1)Women’s height, weight, BMI had no significant difference between two groups (P>0.05).(2)①The number of women who stay up late, smoking, drinking in the observed group was significantly higher than that in the healthy control group(P<0.05). The number of women regular exercise in the observed group was significantly lower than that in the healthy control group (P<0.05).②The number of women with mental tension, long-term depression, suffered a major stimulus, easy to get angry in the observed group was significantly higher than that in the healthy control group (P<0.05).③The number of women who feeling pressure, often on a business trip, College in the observed group was significantly higher than that in the healthy control group (P<0.05). The number of women with the harmonious relationship between the colleagues in the observed group was significantly lower than that in the healthy control group (P<0.05).(3) Comparison of women’s quality of life status in two groups:①The SF-36general health score was711.41±18.91in the observation group, while in the healthy control group was481.23±16.67, which had the statistical significant difference between the2groups (P<0.05).②Physiological health score, mental health score of women in the observation group were significantly lower than in the healthy group (P<0.05).③PF、RP、BP、GH、VT、SF、RE、MH of women in the observation group were significantly lower than in the healthy group (P<0.05).2. Preparation for test(1) The general condition of ratsThe signs appearance, behavior, mental state, feeding, drinking water, hair, breathing, mouth, eyes, nose, ears, size and shape of color of the rats in control group were normal. The rats in the model group were Mao Shusong, reduced activity, dizziness, limb weakness. The signs appearance, behavior, mental state, feeding, drinking water of rats in the Chinese medicine group was equal to the rats in control group. The coat, breathing, mouth, eyes, nose, ears, size and color of rats in the Chinese medicine group were normal.(2) Influence on the estrous cycle ㈠Estrous cycle of rats in blank control group were all the rules of the vaginal smear, the cycle was4-6days, the median was5days.②Thevaginal smear showed estrus cycle change in the model group at the1,2week after injecting CTX, estrous cycle rule disappeared, the cycle was8-17days, the median was11days. The vaginal smears showed little secretion, more scattered in the white blood cells, a small amount of epithelial cells, part of a small amount of keratinized cells and mononuclear cells, diestrus time was5～7days.③The vaginal smear showed estrus cycle change in the Chinese medicine group at the1,2week after injecting CTX, estrous cycle variation rule prolonged, the cycle was6-12days, the median was9days. Vaginal smears showed a small amount of powder in the amount of white blood cells, epithelial cells, a small amount of keratinized cells and mononuclear cells, diestrus time was2~4days.(3) Influence on serum E2, FSH of DOR ratsSerum E2of the rats in the model group at2W,3W,4W decreased significantly, which had significant difference with that in the blank control group (P<0.05or P<0.01). E2of the rats in Chinese Medicine group at3W,4W increased significantly, which had significant differences with that in the model group (P<0.05). While serum FSH was whereas.(4) Influence on ovary index of DOR ratsOvary index of the rats in the model group at2W,3W,4W decreased significantly, which had significant difference with that in the blank control group (P<0.05or P<0.01). Ovary index of the rats in the Chinese Medicine group at1W.2W,3W,4W increased, which had no significant difference with that in the model group (P>0.05)(5) Influence on follicle number of DOR ratsPrimordial follicles of rats in the model group at1W,2W,3W,4W decreased significantly, which had significant difference with that in the blank control group (P<0.01). Primordial follicles of rats in the Chinese Medicine group at1W,2W,3W,4W increased significantly, which had significant difference with that in the model group (P<0.01).Primary follicles, secondary follicles, the mature follicle, atretic follicles at1W,2W,3W,4W decreased significantly, which had significant difference with that in the blank control group (P<0.01). Primary follicles, secondary follicles, the mature follicle, atretic follicles at1W,2W,3W increased significantly, which had significant difference with that in the model group (P<0.05).(6) Influence on ovary morphology of DOR ratsBlank control group:Ovary (1W,2W,3W,4W) can be clearly divided into fcortex and medulla. Cortex had many different sizes, dark staining of primordial follicle, primary follicle, secondary follicle, mature follicle and corpus luteum atresic follicle, and white body. Interstitial gland was rare, and rich in blood vessels. Stromal cells and fibers organized cortex, interstitial was not hemorrhage, edema, degeneration, necrosis, fibrosis Medulla contained abundant nerve, lymphatic and vascular, congestion and degeneration was not found.Model group:Ovary (1W,2W,3W,4W) medullary was atrophy, cortical hyperplasia, fibrosis like change, and the number of atretic follicles increased significantly.Chinese Medicine group:Ovary (1W,2W,3W,4W) medulla was basically normal, no obvious atrophy. Cortical hyperplasia was less. Follicle, corpus luteum was the same as that in blank control group, which was no obvious degeneration, fibrosis and other diseases.3. Effect of Guishen Pill on ovarian function of DOR rats induced by CTX(1)Influence on body weight of DOR rats1week after rats were intraperitoneally injected with CTX:Body weight of rats in model group and other medicine group decreased significantly than that in blank control group (P<0.05).4weeks after given medicine:Body weight of rats in model group decreased significantly than that in blank control group (P<0.05); Body weight of rats in Chinese medicine group and Modern medicine control group increased significantly than that in model group (P<0.01), which had no significant difference with that in blank control group (P>0.05)(2) Changes of the rat estrous cycleEstrous cycle of all rats was normal before given medicine, a period of about4-5d. After given medicine, estrous cycle rats in the other group was changed, stoped at diestrus.4weeks after given medicine, estrous cycle of rats in model group was dysfunction. Rats in Chinese high, medium dose group had complete estrous cycle after administration of14-15d. while in modern medicne group had complete estrous cycle after administration of15-17d. Rats in Chinese medicine low dose group began to appear around the proestrus, after administration of19-20d, duration of8~10d, and appeared estrus before the end of the4week.(3) Influence on serum E2、FSH of DOR rats1week after rats were intraperitoneally injected with CTX:E2of rats in model group and other medicine group decreased significantly than that in blank control group (P<0.05).4weeks after given medicine:E2of rats in Chinese medicine group and modern medicine increased significantly before given medicine (P<0.05), which increased significantly than that in model group (P<0.01); E2of rats in model group decreased significantly than that in blank control group (P<0.01)1week after rats were intraperitoneally injected with CTX:FSH of rats in model group and other medicine group increased significantly than that in blank control group (P<0.01)4weeks after given medicine:FSH of rats in Chinese medicine group and modern medicine decreased significantly before given medicine (P<0.05), which increased significantly than that in model group (P<0.01); FSH of rats in model group increased significantly than that in blank control group (P<0.01).(4) Influence on serum AMH. INHB of DOR rats1week after rats were intraperitoneally injected with CTX:AMH、INHB of rats in model group and other medicine group decreased significantly than that in blank control group (P<0.05)4weeks after given medicine:AMH、INHB of rats in model group in model group decreased significantly than that in blank control group (P<0.01); AMH、 INHB of rats in Chinese medicine group and modern medicine increased significantly than that in model group (P<0.01), which had no significant difference with that in blank control group (P>0.05).(5) Influence on ovary index of DOR rats4weeks after given medicine:Ovary index of rats in model group in model group decreased significantly than that in blank control group (P<0.01); Ovary index of rats in Chinese medicine group and modern medicine increased significantly than that in model group (P<0.01), which had no significant difference with that in blank control group (P>0.05). (6) Influence on follicle number of DOR rats4weeks after given medicine:Number of primordial follicles, primary follicles, secondary follicles, growing follicles and mature follicle in in model group decreased significantly than that in blank control group (P<0.01).Number of primordial follicles in Chinese medicine high, middle dose group decreased significantly than that in model group (P<0.01, P<0.05); Number of primordial follicles in Chinese medicine high dose group had no significant difference with that in blank control group (P>0.05)Number of primary follicles, growing follicles, mature follicles in modle group had significant difference with that in blank control group (P>0.05); Number of primary follicles, growing follicles, mature follicles in Chinese medicing group decreased significantly than that in model group (P<0.05) which had no significant difference with that in blank control group (P>0.05)(7) Influence on ovary morphology of DOR ratsBlank control group:normal ovarian cortex structure, visible at all levels of follicle and corpus luteum distribution, normal morphology, mature follicle size, follicular fluid more.Model group:Ovarian atrophy, reduced growth follicles, follicular atresia more. Chinese high, medium dose group:Growing follicles increased in comparison with the model group, large diameter of the follicles, follicular atresia reduce. Low-dose group:Growing follicles are less visible, primordial follicles. Modern medicine group primary follicles, growing follicles and mature follicles increased, visible.(8) Influence on VEGF, GDF-9of DOR ratsVEGF, GDF-9of rats in model group reduced significantly than that in blank control group (P<0.01), in Chinese medicine group enhanced significantly than that in model group (P<0.01)Conelusions1. The life quality of kidney deficiency and blood stasis of liver stagnation and spleen deficiency type DOR patients decreased significantly. Specific performances in the social function, mental health and overall health of3dimensions damage. The psychological health of DOR patients decreased much more than physical health. The incidence and the social, psychological factors, living habits, cultural level.2. A single intraperitoneal injection of CTX can induce SD rat follicle developmental disorder, sex hormone secretion disorder, ovarian dysfunction. FSH of rats in Chinese medicine group decreased, E2increased, which had no significant difference with that in blank control group. It proved that Guishen Pill can restore the estrous cycle in DOR rats, DOR rats, promote follicular development and ovulation function, improve the DOR serum high gonadotropin low estrogen state, thus recovering the function of ovary.3. Guishen Pill can improve the ovarian reserve function through a variety of ways, specifically in the following aspects:Regulation of the hypothalamic pituitary ovarian axis drop, reduce the secretion of gonadotropin; improve the INHB, AMH level in DOR rats; granulosa cell VEGF levels increased in DOR rats, through paracrine stimulation of thecal endothelial cell proliferation, promote ovarian angiogenesis; can up regulate the expression of GDF-9protein in rats, in order to promote the primordial follicle recruitment, promote the follicular development, ovulation inhibition of follicular atresia, in order to achieve the purpose of ovalution. |
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