| Objective:Endometriosis (EMs)is the disease that the endometrial glands and stroma appear and grow out of the endometrial lining and the muscle of the uterine cavity. There are variety of clinical manifestations of the disease, such as dysmenorrheal,chronic pelvic pain, sexual pain, menstrual abnormalities and infertility. The group of women on childbearing age has the best high incidence rate of the disease, the incidence rate is about 10%～15%.Although the ectopic endometrium is benign on histology, it is with the proliferation, invasion, metastasis and recurrence of malignant behavior and so on. The current clinical treatments for endometriosis are the methods of surgery and drug therapy, but the cost of surgical treatment is very high and the relapsing rate is also high, so we should explore more new and effective drug treatments.In view of the quality of hormone-dependent,western medicine treats the disease with hormone therapy, but the side effects of these drugs are rather worse. Traditional Chinese medicine treatments may be good to treat the disease,as we all know, Chinese medicine has a long history of treatment which not only releases some symptoms effectively, but also has less side effects. So there is great practical significance to explore traditional Chinese medicine treatments for the endometriosis.Xiaoyi Tang is one of Chinese medicine for the endometriosis, and it was used for many years of clinical experience, which was made sure of the effective treatment of endometriosis by many patients.It contains Sanleng, Ezhu, Puhuang, Wulingzhi,Zicao, Dragon Blood, turtle, iron caltrop, Zhejiang Fritillaria, Lujiaoshuang and other Chinese medicine drugs.In this experiment, we built the EMs model of the rats on the abdominal wall muscle with their own endometrial tissue firstly, Xiaoyi Tang then was used to treat the rats.After 3 weeks by using the drugs,we killed the rats and collected their blood and the ectopic endometrial tissue. Then we detected the expression level of EMAb by ELISA, the expression levels of E2,P,FSH and LH by the method of chemiluminescent,and we also detected the expression level of VEGF in the ectopic endometrial tissue by the method of immunohistochemistry. The experiments on Xiaoyi Tang were made in order to approve the effect of treatment for endometriosis and research the mechanisms of the action, which can provide experimental data for clinical application.Method:1.Animal:a total number of 50 SD rats,no mated, sexually mature, females,clean, healthy,weight 180-220g.2. Divided into five groups randomly, sham-operated group 10,model group 10, mifepristone group 10, Xiaoyi Tang high-dose group 10, Xiaoyi Tang low-dose group 10.3. Beside sham-operated group, the rats in other groups were used to establish their own EMs rat transplantation model.After 2 weeks, we tested the ectopic endometrial tissue by the naked eyes.4. At the 15th day after making the Ems model,we began to give the medicine by irrigating gastric. Low-dose group was given dose of medicine 10.5g/kg d, high-dose group was given dose of medicine 21g/kg d, mifepristone group was given 5mg/kg d, sham-operated group and model group were given the liquid of NS by each 4ml/d. All the volume to give the animal was 4ml one time per one day. After 21 days continuously using drugs,the animals were forbidden to eat and drink for 12 hours in the evening,then these animals were killed and their blood and their ectopic endometrial tissue was collected.5.The expression level of EMAb was detected by the method of ELISA. The expression levels of E2, P, FSH and LH was detected by the method of chemiluminescent.The expression level of VEGF in the ectopic endometrial tissue was detected by immunohistochemistry method.6. All data were analyzed statistically by SPSS13.0.Results:1.The method of building the model of endometriosis rat is success by Pathology confirmed through the abdominal transplantation of autologous endometrial,and the survival rate of the transplant tissue is 95%.2.The weight of all the groups was no significantly difference before the operation to make the Ems model.But after 21 days for using drugs,the weight of sham-operated group was obviously higher than that of model group; and the weight of mifepristone group was also higher than that of model group.3. After making model for 2 weeks,the volume of the plantation of every group was no difference in the test.But after 21 days by using drugs,the volume of model group was bigger than mifepristone group, Xiaoyi Tang high-dose group and Xiaoyi Tang low-dose group (P<0.01).The difference between the volume of ectopic endometrium of mifepristone group and Xiaoyi Tang group was statistically significant (P<0.05).The difference between the volume of ectopic endometrium of Xiaoyi Tang low-dose group and Xiaoyi Tang high-dose group was also statistically significant (P<0.05).The effect of Xiaoyi Tang high-dose group reduce the volume didn't have statistics significance between mifepristone group(P>0.05).4. The method by histopathology showed the growth of ectopic endometrial of model group was very well,and that of mifepristone group and Xiaoyi Tang high-dose group was inhibited in which the basic atrophy or endometrial tissue degeneration and necrosis.The size of ectopic endometrial in the model group was bigger than other groups (p<0.05),but the other drug groups was no significant difference each other (p> 0.05).5.The expression of VEGF in ectopic endometrial in model group was higher than sham-operated group,mifepristone group, Xiaoyi Tang low-dose group and Xiaoyi Tang high-dose group(P<0.05).The expression of VEGF in mifepristone group, Xiaoyi Tang low-dose group and Xiaoyi Tang high-dose group were higher than that in the sham-operated group(P<0.01).The expression of VEGF in ectopic endometrial in Xiaoyi Tang low-dose group was higher than that of mifepristone group and Xiaoyi Tang high-dose group(P<0.05).There was no significant difference between mifepristone group and Xiaoyi Tang high-dose group(P>0.05)。6.The expression level of EMAb in the sham-operated group was lower than that of model group, mifepristone group, Xiaoyi Tang high-dose group, Xiaoyi Tang low-dose group(P<0.05).The positive rate of EMAb in the model group was obviously higher than that of sham-operated group, mifepristone group, Xiaoyi Tang high-dose group, Xiaoyi Tang low-dose group(P<0.05).But there were no significant difference between mifepristone group, Xiaoyi Tang high-dose group, Xiaoyi Tang low-dose group each other in the positive rate of EMAb(P>0.05).7.The expression level of estrogen in the sham-operated group was lower than model group,mifepristone group, Xiaoyi Tang high-dose group and Xiaoyi Tang low-dose group(P<0.01).The level of estrogen was no significant difference among sham-operated group, mifepristone group, Xiaoyi Tang low-dose group and Xiaoyi Tang high-dose group(P>0.05).8. The levels of progesterone of mifepristone group were significantly lower than other groups (P<0.01),other groups:model group,sham-operating-group,Xiaoyi Tang low-dose group and Xiaoyi Tang high-dose group didn't exist statistically significant difference each other (P>0.05).9. The expression of FSH and LH in all groups was no statistical difference each other.Conclusion:1.The method of building the model of endometriosis rat was success, the plantation grew well.2.Both Xiaoyi Tang and mifepristone can inhibit the growth of the plantations3. The expression level of VEGF in ectopic endometrial of EMs rat model group was higher than normal endometrial tissue,suggesting that VEGF might play an important role in the pathogenesis of endometriosis.Xiaoyi Tang can reduce the expression level of VEGF in the ectopic endometrial,which might one of the mechanisms of Xiaoyi Tang inhibiting the angiogenesis effectively4.Mifepristone could significantly inhibit the growth of ectopic endometrium. And the main mechanism of its role and its effect was its great impact on anti-progesterone, and reduced the expression of VEGF, too. What's more, mifepristone should have many ways in treatment of endometriosis disease.5. In this experiment,Xiaoyi Tang has not statistically significant effect on E2, P, FSH and LH, but it can effectively reduce the expression of anti-endometrial antibodies,which suggested that the primary mechanism of the treatment for endometriosis was its regulating function on immune by reducing the impact on the body's own antibodies.6.After this test,the possible mechanisms of Xiaoyi Tang might be learned, but there were still many questions.In the future, we should design more scientific and experimental programs to research the traditional Chinese medicine including adding the number of experimental samples. |
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