| Objective:Based on the success of establishing adenomyosis mouse model, through a series of experimental method and effect indicators, evaluating the effects of Xuandansanjie Decoction on adenomyosis mouse model and exploring its mechanism.Methods:The rebirth ICR mice were randomly divided into 5 groups, setting as blank group, model group, western medicine group, low dose and high dose group of Chinese medicine. Blank group is used as control. Model group, western medicine group, low dose and high dose group of Chinese medicine as experimental group. Using the method of oral tamoxifen establish adenomyosis mouse model for mice of experimental group. After three months, we randomly selected 6 mice from model group, western medicine group, low dose and high dose group of Chinese medicine, which were given the suitable dose of distilled water, danazol, low-dose and high-dose Xuandansanjie Decoction. Another six normal mice from the control group were given distilled water. Each group of mice fed once a day, five days a week, continue two months. Pain associated behavioral experiment (tail flick test, hot plate test, prick test machine, test and writhing test acetone) to observe pain sensitivity for every group mice and the analgesic effect of Xuandansanjie Decoction. Two months later the mice were sacrificed. Observe the uterine congestion situation and volume size through naked eyes. Electronic weighing scales the weight of uterus and ovary and caliper measure the diameter of the uterus. Observe mouse uterine lesions nodules under the microscope. Collect mouse uterus made paraffin sections, HE staining to observe morphological changes of the uterine tissue and given pathological score. By immunohistochemistry and RT-PCR to detecte the expression of NGF-β, p75NTR, TrkA proteins and mRNA in uterus and DRG, then statistical processing.Results:1. Pain related behavioral experiments:pain threshold of experimental mice are lower than the control group and more sensitive to pain. Comparative experimental group, pain thresholds of Chinese medicine group and western medicine group were higher than the model group, indicating that the XuanDansanjie Decoction and danazol can alleviate the pain of adenomyosis mice model e and reduce pain sensitivity. The high-dose Chinese medicine group has the most significant change to the pain threshold, suggesting that the effect of high dose Xuandansanjie Decoction is the best.2. Results of drug efficacy:Chinese medicine group and western medicine group all could shrink uterine volume, reduce uterine weight, improved surface hyperemia, reduce the number of nodules and lower pathological scores, suggesting Xuandansanjie Decoction and danazol are valid for adenomyosis Mice and high-dose Xuandansanjie Decoction has the best effect.3. Immunohistochemistry and RT-PCR results:Compared with the control group, the expression levels of NGF-, p75NTR, TrkA protein and mRNA were significantly increased in the uterine tissues. Compared with the experimental group, the expression level in the model group was highest, and the expression level in the high-dose Chinese medicine group was the lowest. In the DRG, compared with the control group, there was no significant difference in the expression of TrkA mRNA between the experimental group and the control group, and the expression level of p75NTR mRNA was significantly increased. Comparison between the experimental group, there was no significant difference in the expression of p75NTR mRNA. The expression level of TrkA mRNA in the model group was highest, and in the high-dose Chinese medicine group was the lowest.Conclusion:1. The animal model of adenomyosis is successful confirmed by oral tamoxifen method, can be used in the study of clinical disease.2. Xuandansanjie Decoction can relieve the adenomyosis model mouse of hyperalgesia.3. NGF and its receptors may involve in the pain mechanism of adenomyosis. Xuandansanjie Decoction relieve adenomyosis induced pain by inhibiting the expression of NGF and its receptor in adenomyosis lesion. |
