| Dysmenorrhea is one of the common clinical manifestations of adenomyosis (AM), seriously affecting the patient’s daily life and work. More and more attention is payed to the study on the treatment of pain and the mechanism of uterine adenomyosis in clinic. In recent years, many domestic and foreign research found that nerve growth factor (NGF) and its receptor has great relationship with the dysmenorrhea caused by AM. "Yi Kun Yi Tong Ping" in treating adenomyosis, in the long-term clinical application has a good therapeutic effect on relieving dysmenorrhea caused by AM. So this research aims to NGF and its receptor as the breakthrough point, through clinical observation and animal experiments to confirm that "Yi Kun Yi Tong Ping" ease AM dysmenorrhea and its mechanism.Objective1 Objective to observe the protective effect and safety of "Yi Kun Yi Tong Ping"on treating dysmenorrhea caused by AM(kidney deficiency and blood stasis Syndrome)by a prospective case control study.2 Objective to observe the mechanism of "Yi Kun Yi Tong Ping", the study use the animal model with adenomyosis, with the NGF and the P75 receptor as the breakthrough point.Method1 Clinical ResearchThe study used the prospective randomized controlled clinical observation, with the "placebo" drug control. And the patients with dysmenorrhea caused by AM of deficiency of kidney and blood stasis were randomly grouped observation method, to found out the "Yi Kun Yi Tong Ping" particles clinical effectiveness and safety.(1) Through the analysis and comparison with the control group, with VAS, COX as evaluation standard, the "Yi Kun Yi Tong Ping" particles are ease to kidney deficiency and blood stasis dysmenorrhea of adenomyosis degree and duration of the evaluation.(2) Through the analysis and comparison with the control group, the "Yi Kun Yi Tong Ping" granule in improving TCM syndrome integral, CA125 and uterine size and other aspects of deficiency of kidney and blood stasis type of adenomyosis of uterus were evaluation.(3) Through the analysis of the treatment group and the control group before and after treatment in patients with blood, urine routine, liver and kidney function, the safety of the "Yi Kun Yi Tong Ping" particles were evaluation.2 Animal ExperimentIn this study, an animal model for the experimental object, changes in cytology and molecular protein levels and receptor NGF. The experiment is divided into two stages, the experiment over a period of 5 months, were divided into 3 groups, each group of 10 mice, divided into blank group, control group and Chinese medicine group.(1)The first stage(the molding stage):Feeding tamoxifen to the ICR mouse was the method to make the model of adenomyosis. The mice were divided into 3 groups, control group and Chinese medicine group were orally administered tamoxifen established mouse model of adenomyosis, blank control group oral saline. Using Histopathological Method under the microscope of muscular layer infiltration to verify whether endothelium model is successful or not, the feasibility of tamoxifen to clear method to establish the ICR mouse model of adenomyosis.(2)The second stage (the experimental stage):After the success of modeling, the blank group and the control group mice were using physiological saline, traditional Chinese medicine groups of mice "Yi Kun Yi Tong Ping" particles stomach intervention. After 8 weeks of treatment, mice were sacrificed, changes before and after the three groups of mice were observed by gross specimen medication, weight of the uterus, tissue pathology; differential expression by real-time fluorescent quantitative PCR method for detection of blot, western three groups of mice uterine NGF and receptor, and to explore:①After treatment, weather there were differences between the traditional Chinese medicine group and the control group mice uterine specimen, uterine weight and histopathology.②After treatment wheThe existence of abnormal high expression of nerve growth factor NGF and its receptor P75 in mouse uterus adenomyosis lesions, and normal mouse uterus contrast analysis.③After treatment, there were differences between the traditional Chinese medicine group and the control group mice uterine expression of NGF and P75.Result1 Clinical Research(1) After 12 weeks of treatment, the VAS scores of the treatment group and the control group, were more decreased than before treatment (P<0.05). Compared with the control group, the VAS scores of the treatment group after treatment was significantly lower than that of the control group (P<0.05), showed that compared with placebo, "Yi Kun Yi Tong Ping" granule can markedly alleviate dysmenorrhea caused by AM.(2) After 12 weeks of treatment, the COX scores of the treatment group and the control group, were more decreased than before treatment (P<0.05). Compared with the control group, the COX scores of the treatment group after treatment was significantly lower than that of the control group (P<0.05), showed that compared with placebo," Yi Kun Yi Tong Ping" granule can markedly alleviate dysmenorrhea caused by AM.(3)After 12 weeks of treatment, the syndrome integral of the treatment group and the control group, were more decreased than before treatment (P<0.05). Compared with the control group, the syndrome integral of the treatment group after treatment was significantly lower than that of the control group (P<0.05), showed that compared with placebo, "Yi Kun Yi Tong Ping" granule can markedly alleviate the syndrome integral caused by AM.(4) After 12 weeks of treatment, the treatment group and the control group had no obvious change of uterine size (P>0.05). It showed that "Yi Kun Yi Tong Ping" Granule had no effect on improving uterine size in patients with AM.(5) After 12 weeks of treatment, the treatment group and the control group had no obvious change of CA125(P>0.05). It showed that "Yi Kun Yi Tong Ping" Granule had no effect on improving CA125 in patients with AM.(6) The treatment group did not find obvious adverse reaction during the treatment. Before and after treatment, blood routine, urine routine, liver and kidney function tests showed no abnormality.2 Animal Experiments(1)Result of Modeling:3 months after taking tamoxifen,one of the model group and blank group mice was killed to observe whether there were successful in the modeling.①The model group mice uterus was bigger than that of the normal control group mice. And the surface of the model mice uterus had the small quantities of adenomyosis nodules.②The model group mice endometrial layer structure disorder, no obvious boundary and muscular layer of uterus, uterine muscular layer visible glandular cells and stromal cells.(2)Result of Experiment①The Chinese medicine group compared with the control group mice, the uterus is increased, and the uterine adenomyosis nodules visible surface.②After treatment, the traditional Chinese medicine group and the control group of mice womb weight had no significant difference (P>0.05).③After treatment, adenomyosis uterine sections of mice still showed muscle layer has obvious glandular and stromal cell invasion, but involvement group compared with the traditional Chinese medicine group is more disorder.④By the Analysis of real time PCR and the westernblot, three groups of mice NGF mRNA values had no significant difference (P>0.05).⑤By the Analysis of real time PCR and the westernblot,the amount of mRNA in mouse uterus adenomyosis model P75 showed increasing trend compared with the blank group, after traditional Chinese medicine treatment and showed a downward trend (P<0.05).Conclusion1 Clinical Research(1) "Yi Kun Yi Tong Ping" granule can obviously relieve dysmenorrhea degree and duration, and also can significantly improve the symptoms of patients with AM.(2)There is no obvious effect of "Yi Kun Yi Tong Ping" Granule on improving uterine size and decreasing the level of serum CA125 in patients withsAM.(3)There is no obvious adverse reaction of "Yi Kun Yi Tong Ping" during the treatment process.2 Animal Experiments(1)There is no obvious effect of "Yi Kun Yi Tong Ping" Granule on improving the uterine size and weight of the AM model mice.(2)There is no obvious effect "Yi Kun Yi Tong Ping" Granule on endometrial glands relief AM model mice and interstitial infiltration of muscle layer, but seemed to slow down the speed of infiltration of interstitial cells of glands.(3) By the analysis of Real-time PCR and Western blot, there is no abnormal high expression of NGF in the AM mouse model uterus. And there is no changes in the expression after treatment of Chinese medicine.(4) By the analysis of Real-time PCR and Western blot, there is abnormal high expression of P75 in the AM model mice uterus. And after treatment of Chinese medicine, the expression of P75 reduces. That shows, Yi Kun Yi Tong Ping relieve the dysmenorrhea by acting P75.
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