Experimental Study On Induced Rats With Tamoxifen And Rhizoma Curcumae Treat Endometriosis

OBJECTION: As so far, treatments to endometriosis include medical therapy, surgical therapy and combination drug and surgical treatment. The main mechanism of medical therapy is medicine could inhibit ovulation. 30% patients would be recurrent after cessation of administration. All present therapies face up to difficulty. The treatment effect is expected to improved.The etiology of endometriosis is still unknown now. Menstruate reflux and implant theory and coelom epithelium metaplasia theory have been accepted widely. There is evidence which manifest that the dense of E₂ where ectopic endothelium implanted increased, which could contribute to the growth of ectpoic lesions. Tamoxifen, which belongs to selected estrogen receptor modulators (SERM) , could block estrogen's function. While Rhizoma curcumae, one kind of traditional Chinese medicine, has autitumor function and could enhance immune power of organism. It had been confirmed that the mechanism of Rhizoma curcumae's antitumor function was to induce cell apoptosis.This experiment would combine one SERM—Tamoxifen and cell apoptosis contributor — Rhizoma curcumae to treat the rats with endometriosis. To observe the therapeutic effect of the medicine on rats with experimental induced endometriosis. It is expected to find one more effect and thorough method that could treat or cure endometriosis. METHOD: Choose 50 female SD rats which weigh 200-250 grammar. Observe the smear of vaginal cell at settled time every day. Select those rats which estrus cycle is 4-5 days and has continuous 2 estrus cycle. Establish rat models with endometriosis during estrus ( the smear of vaginal cell is hyperkerotic cell )according to Jones method.Four weeks after the autotransplantation, take the second paunch to investigate the growth of transplantation and mesure its volume. Eliminate those rats that autotransplantation is not well grow. The rats with peritoneal implants were randomly assigned into 4 groups. One group was control group, the other 3 group were treated respectively with Tamoxifen 5mg/kg fill stomach twice per day, GnRHa(Alarelin) 30ug/kg muscle injection once per day, Tamoxifen 5mg/kg fill stomach twice per day combined Rhizoma curcumae oil lOmg/kg peritoneal injection once per day.After 4 weeks, the rats models were killed after observing continuously the smear of vaginal cell for 6 days. Take the third paunch to observe the shape of ectopic lesions and mesure lesions' diameter and height. Cutting eutopic endometrium and ectopic implants, dipped in 10% formalin, paraffin embed, make 4 u m thick continuous slice, HE stain, observe the pathologic change of eutopic and ectopic endometrium.Evaluate the score of expression levels of vascular endothelial growthfactor(VEGF)> bcl-2 and bax of ectopic implants which were detected by streptavedin-peroxidase. First antigen was substituted by PBS in negative control. Positive control is known positive slice. Observe slice under microscope.All data were showed by x + s and dealed with SPSS 10.0 statistical software. Estimate the ectopic lesion's volume has difference or not between pre-treatment and post-treatment with t test. Estimate the ectopic lesion's expression of VEGF n bcl-2 > bax has difference or not after treatment with analysis of variance(ANOVA). Inspect the difference of those expression existed among groups by q test.RESULTS: 1. The volume of endometrial implants of all treatment group has significant difference between pre-treatment and post-treatment (111. 79 mm3vs89. 60 mm3; 112. 00 mm3 vs 31. 34 mm3; 115. 22 mm3vs 40. 99 mm3) (P<0.05). 2. The volume of lesions of all treatment group markedly smaller compared with control group after treatment (89. 60 mm3 vsl26. 79 mm3; 31. 34 mm3 vsl26. 79 mm3; 40. 99 mm3vsl26. 79 mm3) (PO.05).3. The score of VEGF was significant difference between each treatment group and control group (3.00vs4.30, 1.67vs4.30n 2. 11vs4.30) (PO.05).4. The score of was VEGF markedly difference respectively between Tamoxifen group and GnRHa(Alarelin) group, Tamoxifen group and Tamoxifen+ Rhizoma curcumae oil group (3.00 vs 1. 67; 3.00 vs 2. 11) (PO.05). 5.The ratio of bcl-2/bax in GnRHa group is smallest (0. 371) , the smaller is Tamoxifen+ Rhizoma curcumae oil group (0. 44) . CONCLU SION: After treatment, the volume of endometrial implants of all treatment group has significant difference compare with control group, thevolume of Tamoxifen+ Rhizoma curcumae oil group has significant difference compare with Tamoxifen group. These show that selected estrogen receptor modulators(Tamoxifen) combine cell apoptosis contributor — Rhizoma curcumae could decrease evidencedly the volume of ectopic implants of rats. Lnmunohistochemistry defect manifest that the expression levels VEGF of Tamoxifen+ Rhizoma curcumae oil group decrease and apoptosis increase compare with Tamoxifen group, which show that combined treatment has some experimental effect on deal with induced rats model. It is worth that more extensive investigation on mechanism and the value of clinical practice.