| ObjectiveStudy on the effect of kidney tonifying method of primary ovarian insufficiency (POI) in patients with anti Mullerian hormone (AMH) level of influence. Objective to explore the kidney filled fine treatment of the POI effect. Study on the method of tonifying kidney of POI mice of AMH signal transduction pathway (AMH/ALK2/Smad1) effect, explore the mechanism of kidney tonifying method in the treatment of POI.MethodClinical study:a total of 2014.3～2014.12 in 80 cases of the First Affiliated Hospital of Guangzhou University of Chinese Medicine, Department of gynaecology outpatients with POI diagnosis standard patients, age range 22～39 years, mean age was 30 years. According to the serum levels of FSH patients were divided into 4 groups:group A(10IU/L<FSH≤20IU/L)、group B(20IU/L<FSH≤30IU/L)、group C(30IU/L<FSH≤40IU/L)、group D (FSH>40IU/L),20 cases in each group, including traditional Chinese medicine group and Western medicine group of 10 cases, Chinese medicine group with kidney fine treatment, with Zuogui pill for basic decoction, western medicine group were treated with kidney tonifying method of sequential therapy of estrogen and progesterone (progynova lmg* 21 days, QD, on the twelfth day Huangtitong Jiaowan 100mg, bid*10 days). The traditional Chinese medicine group, western medicine group were treated for 6 months. The improvement of the symptom score were observed in patients with menstrual and systemic symptoms, changes of chemiluminescent immunoassay in detection of serum AMH, FSH levels by electrochemistry. The women in the age group of 21～39 and 20 healthy volunteers as normal group.Experimental study:8 weeks old BALB/C female mice were 40, were randomly divided into model group, Zuogui pill group, estradiol valerate group, blank control group. Another female BALB/C mice aged 9 in December only to the elderly group.0.03mg bearing mouse zona pellucida 3 peptide (ZP3) in the solution of 0.15ml mice after subcutaneous injection of double feet and abdominal cavity, to establish an animal model of POI. After the success of modeling, the drug group were treated with Zuogui pill 2.7g/(Kg · d),0.15mg/(estradiol valerate Kg · d) by intragastric administration of distilled water, model group and blank control group were given the same amount of. Gavage for 4 week, the weekly rest day. Serum and ovarian tissue of mice were observed in the follicular growth, HE staining, detection of serum AMH, FSH levels of mice electrochemical luminescence method, the expression of immunohistochemical method and Westen blot method to detect AMH, ALK2, Smad1 protein in ovarian tissue.Results:(1) With the increase of age, the serum level of AMH in normal women of childbearing age gradually decreased, while the level of FSH increased gradually.(2) After treatment, POI patients with hypomenorrhea, of late, has no regular, amenorrhea has been significantly improved, the total effective rate of treatment was 75%, the kidney tonifying method the treatment efficiency was 72.5%. A group, B group, C group, D group were 80%,70%,80%,60%, for better effect, that tonifying the kidney in patients with early POI menstrual improvement and replenishing essence method.(3) Observed in patients with POI, clinical symptoms such as insomnia, depression is not happy, in group C, patients in group D also showed Hongrehanchu, tired body fatigue, sexual desire is low, the effect of kidney tonifying method after the treatment, the frequency of the main symptoms were reduced.(4) Compared with before treatment, after treatment of tonifying kidney, serum AMH levels in patients with POI increased, A, B, D three group increased significantly, and the effect of traditional Chinese medicine is better in group A; the serum levels of FSH were decreased, B, C, D three group decreased more significantly.(5) Mice injected with ZP3 solution can be successfully established animal model of immune POI mice, showed the estrous cycle disorder, vaginal smears to see a small amount of white blood cells, epithelial cells. Zuogui pill treatment, the estrous cycle is regular, in estrus, vaginal smear showed a large number of keratinized epithelial cells. HE staining showed that in the model group, the ratio of growing follicles in ovaries was significantly reduced (P<0.01), by Zuogui Pill in mice after intragastric administration, the follicle growth rate increased, the difference was statistically significant (P<0.05)(6) It was found to that increase with age, rats, serum AMH level decreased, FSH level increased. Compared with the model group,. Zuogui pill group serum AMH levels were significantly increased, and better than the progynova group (P<0.05), the FSH level was significantly decreased, compared with the observation group, no significant difference.(7) Immunohistochemical method and Western blot method showed that, AMH, ALK2, Smad1 protein was expressed in cytoplasm of granulosa cells. Aged rats ovarian AMH, ALK2, Smad1 protein expression was significantly lower than that of the control group. Compared with the model group, estradiol valerate group, Zuogui pill group mice ovarian AMH, Smad1 protein expression were increased (Western blot detection shows, Zuogui pill group was better than that of progynova group), the expression of ALK2 protein was also increased (Western blot assay showed that the detection of progynova group was better than that of Zuogui pill group).Conclusion1. Effect of kidney tonifying method can improve POI patients with menstruation and systemic symptoms, can reduce the level of serum FSH, increase the level of AMH, progress and intervention of POI.2. Ovarian lower expression of AMH protein may be one of the causes of POI, and may influence the effect of AMH/ALK2/Smad1 signal transduction, primordial follicle recruitment, so that excessive consumption of follicle.3. The effect of kidney tonifying recipe of Zuogui pill can up regulate the expression of AMH protein in POI mouse ovary, possibly by affecting the AMH/ALK2/Smad1 signal transduction, inhibiting too fast raise and consumption follicular, improving the ovarian function,... |
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