Mechanisms Of Bushen Shugan Recipe In Regulating The Hypothalamus-Pituitary-Ovarian Axis Of Anorexia Nervosa's Stress Rat Models

Objective:1) To investigate the methods of modeling anorexia nervosa(AN). To establish the modeling methods of AN's stress models in this study.2) To observe the effects of Bushen Shugan Recipe in regulating the Hypothalamus-Pituitary-Ovarian axis of anorexia nervosa's stress rat models. To investigate the regulating mechanisms.Methods:1) We searched literatures about anorexia nervosa and its animal models and summarized the classic modeling methods. We chose the stress models, which in accordance with our studies.2)We established the modeling methods by experiments, which were improved slightly on the basis of the original methods and confirmed by vaginal cytology smear. The smears showed that the stress models'oestrous cycles were disordered and their diestrus were lengthened, which was similar to AN patients'symptom——amenorrhea.3) Animal experiments:40 Wistar female rats, weighing 160-180g, were chosen from≥50 rats, by vaginal cytology smear. Those 40 rats had at least 2 continuous normal oestrous cycles and were used in the experiments. Dividing the chosen rats into 4 groups according to the random digits table——the normal control group, the model group, the placebo group and the Bushen Shugan Recipe group(that is the Chinese herbs group). There were 10 rats in each group and others were eliminated. From the frist day of the experiments, the nomal control group were given physiological saline by intragastric administration. The other three groups were induced the stress models by the steps of separation, diet restriction and constraint. At the same time, the placebo group were given physiological saline by intragastric administration. While the Chinese herbs group were given Bushen Shugan Recipe's decoction by intragastric administration. Observe the vaginal cytology smear everyday.4) During the rats'diestrums and on the 35th day of the treatment, the body weights and the oestrous cycles of the 4 groups were compared. The levels of serum estradiol (E2), hypophysis luteotrophic hormone (LH), hypophysis follicle stimulating hormone (FSH) and hypothalamusβ-endorphin (P-EP) were detected with the radio-immunity, while the levels of serum corticosterone (CORT), hypothalamus dopamine (DA), hypothalamus noradrenaline (NE) and hypothalamus serotonin (5-HT) were detected with euzymelinked immunosorbent assay.Results:1) In our experiments, rats were induced the AN's stress models by the steps of separation, diet restriction and constraint. Vaginal cytology smears showed that the models' oestrous cycles were disordered and their diestrus were lengthened, which is similar to AN patients'symptom——amenorrhea. The models could be used in our studies.2)The body weights of the model group, the placebo group and the Chinese herbs group were significantly lower than the normal control group's (P<0.01). The Chinese herbs group's body weights were significantly higher than the placebo group's (P<0.01), and higher than the model group.3) The rats' oestrous cycles:we computed the rates of cycle disordering of the 20th day, the 25th day and the 30th day. The rates of the Chinese herbs group were higher than the model group and the placebo group.4) Organ coefficient: there were no obviously significance among the four groups'hypothalamus coefficient, hypophysis coefficient or ovaryv coefficient. The placebo group's uterus coefficient were obviously lower than the nomal control group's (P<0.05). The placebo group's adrenal glands coefficient were significantly higher than the nomal control group and the model group's (P<0.01).5) The model group's hypophysis FSH were obviously lower than the nomal control group's (P<0.05).The placebo group's hypophysis FSH were significantly lower than the nomal control group's (P<0.01). The Chinese herbs group's hypophysis FSH were higher than the the model group and the placebo group's. The model group's hypophysis LH were significantly lower than the the nomal control group's (P<0.01). The Chinese herbs group's hypophysis LH were obviously higher than the model group's (P<0.05), and higher than the placebo group's.6)The model group and the placebo group's serum E2 were obviously lower than the nomal control group's (P<0.05).The Chinese herbs group's serum E2 were obviously higher than the model group's (P<0.05), and higher than the placebo group's. The model group' serum CORT were obviously higher than the nomal control group's (P<0.05). The placebo group's serum CORT were significantly higher than the nomal control group's (P<0.01). The Chinese herbs group's serum CORT were obviously lower than the model group'(P<0.05), and significantly lower than the placebo group's (P<0.01).7)The model group and the placebo group's hypothalamus P-EP were significantly higher than the nomal control group's (P<0.01). The Chinese herbs group's hypothalamusβ-EP were obviously lower than the the placebo group's(P<0.05), and lower than the model group's.8) The placebo group's hypothalamus DA were obviously higher than the nomal control group's (P<0.05). The nomal control group's hypothalamus NE were obviously higher than the model group and the placebo group's (P<0.05).There were no obviously significance among the four groups' hypothalamus 5-HT.Conclusion:1) In our experiments, rats were induced the AN's stress models by the steps of separation, diet restriction and constraint. The models could be used in our studies. 2) Bushen Shugan Recipe increased body weights of the rats, postponed the disordering of the rats'oestrous cycle, increased uterus coefficient and reduced adrenal glands coefficient.3) Bushen Shugan Recipe increased the hypophysis LH and serum E2 of the AN's stress rat models obviously, and decreased their serum CORT and hypothalamusβ-EP obviously. That's the paths of Bushen Shugan Recipe in regulating the HPOA.