| Objective: Using adult female rats underwent bilateral ovariectomy surgery to simulate the perimenopausal syndrome. Give the chinese medicine of JPYK and estradiol valerate intervene in the model. Observation of the rats estrous cycle, rectal temperature, levels of serum sex hormones and calcitonin gene-related peptide(CGRP), uterus weight coefficient, uteru tissue morphology and metrology explores the relationship between CGRP and perimenopausal hot flashes, as well as therapeutic effects and the mechanism in rats of traditional Chinese medicine for perimenopausal syndrome, provide the experimental basis for the theoretical basis of perimenopausal syndrome.Methods: 30 healthy and clean, non-pregnant adult female SD rats of weight of 200-250 g were adaptive feeding for 1 week. They were randomly divided into the sham operation group of 6 rats and the model group of 24 rats. After fasting 12 h, model group was underwent bilateral oophorectomy, while sham operation group was removed only a small amount of fat around the bilateral ovaries. After 5 days of the operation, rats whose vaginal exfoliated cell cornification index<50% and with estrous cycle dysfunction are successful as model. After the success of modeling, the model rats were randomly divided into model control group, chinese medicine treatment group, western medicine treatment group, the combination therapy group per 6 rats. Chinese medicine treatment group were given JPYK decoction of 2ml per rat, western medicine treatment group were given estradiol valerate water solution of 2ml per rat, the combination therapy group were given JPYK decoction of 1ml and estradiol valerate water solution of 1ml per rat, and the sham operation group and model control group were given equal volume of distilled water. Test the estrus cycle and rectal temperature of rats regularly. After 2 months of treatment, fasting 12 h, body weight of the rats were measured under anesthesia, decapitate blood, separate serum to determine levels of serum sex hormone and CGRP, remove the uterus to calculate the uterus weight coefficient, cut a part of uterus tissues and observe changes of uterus morphology and metrology under the light microscope.Results: 1 Changes of estrous cycle in rats Pre-modeled: In experiment study, the activity of pre-modeled rats were multifarious, their responds were intelligent, the fur was smooth, and estrous cycle was regular(4-5days/times). Post-modeled: The model group rats responded slowly to stimulate, decreased activity frequency, grew darkness of the fur, while estrous cycle disordered. After treatment: Chinese medicine treatment group, western medicine treatment group and the combination therapy group rats was frequent of the activity, sensitive response to stimulate, sheen of the fur, while estrous cycle became normal gradually. 2 Changes of rectal temperature in rats From figure 1, it can be seen that the interaction existed between different time points and groups, from the rectal temperature trends over time, and the difference was statistically significant between each group. The model group rats rectal temperature compared with the sham operation group there was increased significantly after ovariectomy, the difference was statistically significant(P<0.01). Rectal temperature of the treatment group rats were significantly lower than the model group, the difference was statistically significant(P<0.01). There was no significant difference among chinese medicine treatment group, western medicine treatment group and the combination therapy group in the rats rectal temperature(P>0.05). 3 Changes of serum sex hormone levels in rats Serum sex hormone levels of the model control group rats FSH, LH increased and E2 decreased, compared the sham operation group with significant difference(P<0.01). Serum sex hormone levels of the treatment group rats FSH, LH decreased and E2 increased, compared the model control group with significant difference(P<0.01). Serum sex hormone levels of the combination therapy group rats E2 increased, compared the western medicine treatment group with significant difference(P<0.05). Serum sex hormone levels of FSH had no significant difference between he combination therapy group and chinese medicine treatment group(P>0.05). There was no significant difference between the treatment of each group rats in serum sex hormone levels of LH(P>0.05). 4 Changes of calcitonin gene-related peptide level in rats Calcitonin gene-related peptide levels of the model control rats were significantly higher than the sham operation group, the difference was statistically significant(P<0.01). Calcitonin gene-related peptide levels of the treatment group rats were significantly lower than the model control group, the difference was statistically significant(P<0.01). Calcitonin gene-related peptide levels of the combination therapy group rats decreased, compared the western medicine treatment group with significant difference(P<0.05). 5 Changes of uterus weight coefficient in rats The uterus weight coefficient of the model control group rats decreased obviously. There was significant difference compared with the sham operation group(P<0.01). The uterus weight coefficient of the treatment group significantly increased, compared significant differences with the model control group(P <0.01). There was no significant difference between the combination therapy group and the sham operation group in the rats uterus weight coefficient(P>0.05). 6 Changes of uterus morphology and metrology 6.1 To observe the pathological morphology of uterus Observation under the light microscope: Endometrial epithelial cells and glandular epithelial cells of the sham operation group were highcolumn, glandular tissue denser, glandular cavity sizes, muscular fibers arranged regularly consistent, the serosal layer was smooth. The model control group endometrium obviously atrophied and thinned, glandular tissue decreased, glandular cells mostly were low columnar, muscle layer becomes thinner, muscular fiber arranged disorder, the serosa was not smooth. Compared with the model group, chinese medicine treatment group endometrium thickened mildly, glandular tissue increased, muscular fiber arranged regularly, while the serosa became smooth. Compared with the model group, the endometrial hyperplasia of western medicine treatment group and the combination therapy group was obvious, endometrial epithelial cells was high columnar, glandular tissue denser, muscle fibers were arranged in rules, serosa was smooth. 6.2 The results of endometrial morphometry Compared with the sham operation group, the model control group endometrial thickness and uterine wall thickness decreased significantly, the difference was statistically significant(P<0.01). Compared with model group, uterus morphology measurement indexes of the treatment group thickened obviously, the difference was statistically significant(P<0.01).Conclusions: 1 Menopausal hot flashes, endocrine disorders, histopathological changes demonstrated that perimenopausal syndrome animal model is established successfully. 2 JPYK decoction can improve the general condition of perimenopausal rats, shorten estrous cycle, promote them to recover normally. Changes of serum sex hormone levels, the weight coefficient of uterus, uterus tissue morphology and metrology of the treatment group rats show that has the advantages of chinese medicine in improving endocrine and ovarian function, while the treatment medicine can lower the rectal temperature and the leves of calcitonin gene-related peptide to improve hot flashes and other symptoms of perimenopause in rats. That establishes the experimental basis for the clinical treatment of perimenopausal syndrome. 3 By statistical analysis of the treatment group, the combination therapy group has a better effect. The treatment group can improve perimenopausal symptoms in rats, while the results show that the combination therapy group can reduce the amount of hormones and related adverse reaction. So modern medicine should be closely combined with traditional chinese medicine theory. we should research perimenopausal syndrome from different stages, multi-channel, multi-level to pay equal emphasis on prevention and treatment. |
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