| Premature Ovarian Failure (POF) is caused by many factors. It occurred in women after adolescence but before 40 yeas old. It is characterised by secondary amenorrhoea, hypo-estrogenism and elevated gonadotrophins serum levels. According to these prevalence data, POF affect 1-3% of women. POF can cause a series of symptoms of menopause and sterility. The treatment of POF is very difficult and it seriously affects women's health. My tutor, Professor Zhang Yu-zhen considers the pathogeny of POF is deficiency of kidney and spleen, liver-qi stagnation and blood stasis. We treat POF with Jia Jian Gui Shen Pill, which can tonify kidney and spleen, regulate liver and remove blood stasis. The effect is good. From clinical and experimental research, we study the mechanism of Jia Jian Gui Shen Pill from cell aoptosis respect.1 Literature StudiesThe etiology of POF is unclear, perhaps because of the decrease of ovum, ovum atresia and the damage of ovum after birth. Many scholars recognize that POF is associated to autoimmune disorders, inheritance factors, abnormity of gonadotropin and its receptors, metabolize or infection or damnification factors etc. Among all these factors, autoimmune disorders are the main cause, account for about 20% of POF.The proliferation and differentiation of granulosa cell in ovum is the postulate of ovum development and mature. The apoptosis of granulosa cell is the premise of ovum atresia. The main factors that involves the mechanism of it concludes hormone, cytokine that can regulate ovum apoptosis and some genes.The treatment of modern medicine includes hormone replacement treatment (HRT), inducing ovulation protocol, donation of ovum and artificial fertilization, immune therapy and transplantation of ovary. These methods have such problems: 1 these treatments above have no satisfactory effect. 2 the period of HRT treatment is long and has side effects,which have potential danger to breast and some other estrogen target organs.In traditional Chinese medicine, POF is included in amenorrhea, hypomenorrhea, anemia, early menopause and sterility. Presently traditional Chinese medicine treatment of POF includes such methods: treatment by differentiation of syndromes, fixed formula and Chinese traditional patent medicine.2 Clinical StudiesObjective: 30 women with POF are chosen as therapeutic group. 8 women with POF, which had taken HRT treatment for 6 months, are chosen as control group. Normal group includes 18 healthy and grow-up women, whose periods are regular.Method: The therapeutic group is treated with Jia Jian Gui Shen Pill. Symptoms are observed pre and post treatment. Electrochemistry radiation method has been used to examine follicle-stimulating hormone (FSH), luteinising hormone (LH) and estradiol (E2) pre and post treatment. Tumor necrosis factor alpha (TNF-a) and interleukin-2 (IL-2) have been checked by enzyme-linked immunosorbent assay (ELISA) pre and post treatment. The blood flow parametes involving PSV, PI and RI of both ovary arteries are measured by pelvic ultrasonography.Results: Preceding treatment, symptoms of flush and sweating, fatigue and irascibility, and vaginal dryness are obvious. But after treatment these symptoms are significantly better than before (P<0.01, P<0.05). Symptoms such as weakness of waist and knees, asomnia, heartthrob, stuffiness, and night sweat are ameliorated significantly (P<0.05). The integral of symptoms is decreased significantly compared with pre-treatment (P<0.01).The gonadotropin level of POF patients is not notably different from before treatment. FSH level in the therapeutic group is decreased notably after treatment compared with before (P<0.01), whereas LH level is not decresed notably (P>0.05). Estradiol level increased significantly compared with pre-treatment (P<0.05). Gonadotropin level in blood serum has not changed significantly compared with before in control group (P>0.05). After treatment Estradiol level in therapeutic group is higher significantly compared with control group (P<0.01). But FSH and LH level have no difference (P>0.05).TNF-a level in POF patients is not different compared with normal group (P>0.05). IL-2 level is higher than normal group (P<0.05). The level of TNF-a and IL-2 is not different between pre and after treatment (P>0.05).15 patients were checked by pelvic ultrasonography. The blood flow of both ovary arteries in POF patients is fewer than normal person. PSV in POF patients is lower than normal group (P<0.01). There is no difference of PI between POF and normal group(p>0.05). RI in POF group is higher than normal group (P<0.05).After treating with Jia Jian Gui Shen Pill, 6 patients were healed, 7 patients have good effect, 13 patients have effect and only 5 patients have bad or no effect. So the overall efficiency is 86.67%, and there were 3 sterile patients become pregnant.3 Experimental StudiesWe cultured Wistar rat's granulosa cell and protracte the growth curve of granulosa cell. Using serum pharmacological method, we observed the influence of Jia Jian Gui Shen Pill pastille serum on rat's granulosa cell apoptosis. The control class is Zi Shen Yu Tai pill pastille serum and human menopausal gonadotropin (HMG). Estradiol and progesterone level in culture medium were checked by enzyme-linked immunosorbent assay (ELISA). The percentage of apoptosis was checked by cytometry. BCL-2 protein expression was checked by immunofluorescence.Results: The amount of granulosa cells decreased after separation, especially from 1 to 3 days, and stabilized after that. Estradiol and progesterone level in all groups are not different before adding pastille serum (P>0.05). 6 hours after adding pastille serum estradiol in Jia Jian Gui Shen Pill pastille serum with high dosage is higher than in blank serum and HMG group (P<0.05,P<0.01). Estradiol in Zi Shen Yu Tai pill pastille serum group with medium dosage is different significantly from HMG group (P<0.05). Progesterone in high dosage of Jia Jian Gui Shen Pill pastille serum group is different significantly from blank group (P<0.05).Apoptosis percentage of the lower dosage of Jia Jian Gui Shen Pill pastille serum group is lower significantly than other groups. It is different from groups of the high dosage of Jia Jian Gui Shen Pill pastille serum, the medium group, Zi Shen Yu Tai pill pastille serum groups and the blank group (P<0.01, P<0.05).BCL-2 protein expression of Jia Jian Gui Shen Pill pastille serum groups are different significantly from the blank serum group (P<0.01, P<0.05). The high and medium dosages of Zi Shen Yu Tai pill pastille serum groups are different significantly from blank serum group (P<0.05). The lower dosage of Jia Jian Gui Shen Pill pastille serum group is different significantly with the high and lower dosage of Zi Shen Yu Tai pill pastille serum groups (P<0.05). Conclusion: Chinese herbs that have the effect of tonifying kidney and spleen, regulate liver and removing blood stasis can promote BCL-2 protein expression. So the cell apoptosis is restrained. |
PDF Full Text Request