| Objective: Endometriosis occurs in the female reproductive system, it is a benign disease with invasive. The Endometriosis refers to a growth activity of endometrial tissue outside the uterine cavity and abnormal growth, it is Often causes patients with chronic pelvic pain, dysmenorrhea, dyspareunia, menstrual disorders, bowel pain, infertility. Endometriosis is a common gynecological disease, but so far it has not been fully clarifiedthe. The disease is full of confusion but encourage people to explore. At present, Endometriosis drug therapy treatments mainly uses hormone drugs, but the side effects of these drugs are large and easy to relapse after discontinuation. Many unanswered questions need to solve. Therefore, developing new resistance to endometriosis drugs has a great significance. Because Chinese medicine has its unique medicinal property theories and exact the clinical curative effects, more and more clinicians pay attention to it, all these making Chinese herbal medicine treatment endometriosis have broad application prospect.Saponin is diosgenin and sugar composition. Aglycone spiral sterane class as steroidal saponins saponin. The sugar composition of saponins common glucose, glucuronic acid, galacturonic acid, rhamnose, galactose, arabinose, and so on. Triterpenoid saponin is diosgenin as triterpenoid saponins. The types are more than steroidal saponins, the Triterpenoid saponindistribution is very broad in nature. It is mainly exist in Leguminosae, Araliaceae, Polygalaceae, Cucurbitaceae. Such as ginseng, cohosh, panax, and licorice contains triterpenoid saponins. In recent years, with the fast development of technology, such as chromatography, spectral, activity testing, for Triterpenoid saponins research have some breakthroughs, its many important biological activity and pharmacological activities has been confirmed. For example, Anti-cancer, immunomodulatory, antipyretic, analgesia and antiulcer, allergy, antivirus, hypolycemic, prevention and cure disease of heart head blood-vessel, etc. However, triterpenoid saponin treatment of endometriosis has been reported rarely.In this study, we use autologous transplantation to establish rats endometriosis models, utilize Triterpenoid saponins to treat the rat endometriosis. By observating the change of IL-6, TNF-a, IFN-γ, NF-κB in the rats, discussing the mechanism of Triterpenoid saponins for endometriosis, in order to provide experimental basis for clinical treatment and a effective strategies for endometriosis.Methods: Estrous period without measurement, but in accordance with the induced animal model of endometriosis to establish rats endometriosis animal model. Four weeks after modeling, 40 become the model rats and they were randomly divided into five groups(n=8): Model control group, Mifepristone influences group, Triterpenoid saponins high, medium and low dose group. In accordance with administration and the grouping method, the 40 rats are gavaged for 28 days then collect the specimens for experiment.1. Measuring the volume of ectopic endometrial V2, compared with volume V1 that after the success of mould maker ectopic endometrium, observe the inhibition of the drugs on the ectopic endometrial. Calculate the drugs on the inhibition rate of ectopic endometrial. Calculation formula: Inhibition rate=( V1- V2)/ V1×100%.2. Collect rats serum and abdominal liquid: adopt double antibody sandwich enzyme-linked immunosorbent analysis(ABC-ELISA) to detect the rats interferon-γ(IFN-γ) level in serum and abdominal liquid.3.Cut each group ectopic endometrium, HE staining check the pathological morphology of rats endometriosis.PV immunohistochemistry method used to detect the expression of NF-κB in the ectopic endometrial tissues.4. Take each ectopic endometrium 100mg to extract total RNA by Trizol reagent, adopt RT– PCR to detect the IL - 6 and TNF - amRNA expression in ectopic tissues. 5. The results were expressed as the mean±S.E.M.On-wayANOVA was used to compare the outcomes. It is a statistieally signifieant difference(P<0.05).Results:1. Rat model of endometriosis successfully established, 72.7% into the mold. In each group, before treatment the ectopic endometrium was no significant difference in volume(P>0.05).2. After treatment, the drug group ectopic endometrium volume reduced to a vary degrees, endometrial vesicles liquid reduce or disappear. Compared with the control group, each group was less than ectopic endometrial volume model group, pairwise comparisons have significant differences(P<0.05). The rest between groups pairwise comparisons have significant differences(P<0.05).3. ELISA test was used to examined the serum and peritoneal fluid levels of IFN-γafter treatment. The IFN-γcontent in the model control groups are below the drug group, pairwise comparisons have significant differences(P<0.05). Triterpenoid saponins high-dosage groups have the strongest effect, they have the highest content of IFN-γ; compared with Other each drug group, Pairwise comparisons have significant differences(P<0.05); Mifepristone group and triterpenoid saponins in the medium dose group, no significant differences (P>0.05); The rest of the drug group Pairwise comparisons have significant differences(P<0.05).4. Pathological morphology observation of endometriosis in rats under optical microscope: Model group ectopic endometrium grow well. In addition to the low dose group,the other groups showed different degrees of ectopic endometrial epithelial glands to reduce atrophy and thinning or atrophy.5. PV immunohistochemical method to detect the NF -κB expression in each group endometriosis that after treatment: Testing positive for glandular epithelium cells in the cell nucleus appear brown particles dyeing. Each group in the ectopic endometrial glandular epithelial cells show varying degrees of cytoplasmic staining, nuclear staining is also visible. The expression of NF-κB in the model control group is stronger than the others drug group. Pairwise comparisons have significant differences(P<0.05). Triterpenoid saponins high-dosage groups have the strongest effect, expression of NF-κB is the weakest. Compared with Other each drug group, pairwise comparisons have significant differences(P<0.05); Mifepristone group and triterpenoid saponins in the medium dose group, the expression of NF-κB no significant differences (P>0.05).6. RT - PCR method to detect IL - 6 and TNF - a mRNA expression in each rat ectopic endometrium organization that after treatment. IL - 6 and TNF - a mRNA expression in ectopic endometrium organization of model control group is stronger than the others drug group.Pairwise comparisons have significant differences(P<0.05); each drug group have different levels that inhibited IL-6 and TNF-a mRNA expression in the ectopic endometrial , Triterpenoid saponins high-dosage groups have the strongest effect, compared with Other drug group, Pairwise comparisons have significant differences(P<0.05); the others drug group Pairwise comparisons have significant differences(P<0.05).Conclusion: Triterpenoid saponins have a obviously degradation function on the rat endometrium displacement, the mechanism is probably that Triterpenoid saponins can inhibit NF -κB activity, So can make cytokines interferonγsecretion increased, the expression of IL-6, TNF-a decline concerned, then inhibit the formation and development of lesions. |
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