The Research Of Hypoxia Theory On Pathogenesis Of Endometriosis

Part ? The expression and biological relevance of ATPase inhibitory factor 1 in reproductive systemObjective:To determine IF1 differential expression and associated withhypoxia and glycolysis in tissues of reproductive system.Methods: The expression of IF1, HIF-1a, GLUT1, LDHA, and PDK1 assessed by immunohistochemistry in all the samples, including different cohort of human primary cancers and corresponding normal tissues in the ovary(n=21,15),the cervix(n=30,18), the fallopian tube(n=19,11), the endometrium(n=30,38); decidua and chorionic villi(n=30); normal myometrium and leiomyoma(n=24,30); the eutopic endometrium(n=33) and ectopic lesions(n=33) from women with endometriosis.Quantitative analysis of IF1 protein and mRNA in endometriosis by western bolt andreal time RT-PCR.Result(s): We show that IF1 is highly overexpressed in malignant tumor of reproduvtive system(carcinomas of ovary, cervix, fallopian tube, and endometrium),embryonic tissues(decidua and chorionic villi ), and ectopic lesions of endometriosis.HIF-1a and the key enzymes of glycolysis, including GLUT1, PDK1, and LDHA are also affluent in this tissues. In contrast, IF1 and markers of the hypoxic response were negligible or low expressed in leiomyoma and normal tissues in reproductive system,besides the endometrium.Conclusion(s): IF1 expression has relevance to malignant transformation, cellular proliferration, and hypoxic response in reproductive system.Part ? The Effect of Hypoxic preconditioning on the survival capacity of endometrial mesenchymal stem cellsObjective: This study explored the hypothesis that hypoxic preconditioning(HPC) on endometrium mesenchymal stem cells(EnSCs) before shedding with retrograde menstruation into pelvic cavity is a prosurvival mechanisms to lethal anaerobic damage which may help the formation of endometriosis lesions.Methods:Cells isolated from human uterus endometrium were expanded and characterised by flow cytometry. Detected the surface epitope pro file,cloning forming,and adipogenicdifferentiation of cells between cultured under 21% 02 and 1% 02.The expression of HIF-la and LC3 in cells under 1% 02 for 24h, 48h, and 72h were detected by western blot, respectively. The level of VEGF in supernatant were measure by ELISA. Cultured cells under 1% 02 or 21%02 for 48 h, then transfered to a condition with 0.1%O2 and serum-free culture media. Survival rate of EnSCs in two groups were measured.Result(s):Cells were positive for mesenchymal markers (CD90, CD73, CD105, CD13,CD44), cell adhesion molecules (CD 166, HLA-ABC), and were negative for hematopoietic lineage markers(CD34, CD45, CD14, HLA-DR, CD19). There is no difference on cells surface epitope profile,adipogenic differentiation, and cloning formingpretreatedwith 21% 02 and 1% 02 for 48h. Culture of cells exposed to 1%02had no effect on cells proliferation activity. Cultured cells under hypoxic conditions (1% 02) for 24 h,expression levels of hypoxia-inducible factor 1-a(HIF1-?) elevated and remained constant until 72h. Expression of VEGF in superant and LC3 ? in cells were significantly increased under hypoxia for 48h. Survival rate of EnSCs cultured under 0.1% 02 and serum -free condition is higher in HPC treatment than non- HPC.Conclusion(s):HPC for 48h before exposure to a lethal anaerobic and serum -free condition can give EnSCs a state of adaptation and prosurvival than in normoxia environment by increased expression of pro-survival and pro-angiogenic factors,which may contribute to the survival of refluxed endometrial cells at ectopic sites.Part ? Effect of hypobaric hypoxic preconditioning on surgically-induced endometriosis by allotransplant of uterine tissue in ratObjective: To determine the effects of hypobaric hypoxia prectreatment on surgically induced endometriosis in a rat model.Methods: Six rats were randomized divided into two groups and treated with hypoxia(atmospheric pressure of 6000 m altitude, 8% O2) or normoxia(ordinary presure, 21% O2) for 6h respectively. Then the uterine endometrium of six rats were intraperitoneal implanted into two groups of estrogen-treated ovariectomized Lewis rat, accordingly.The growth and quality of the implants were measured, the expression of proliferation, apoptosis and angiogenesis-associated markers (Ki67,TUNEL, VEGF, CD31) were assessed using immunohistochemistry, the value of VEGF and TNFa were detected in plasma and peritneal fluid separately, and the expression of HIF-la and VEGF were evaluated by Western blot and real-time PCR analyses respectively.Result(s): The volume of the implants in the hypoxic pretreatment group was significantly increased compared with the normoxia group(P<0.05). There were higher expression of Ki67, CD31, VEGF, and HIF-la and lower apoptotic changes in the hypoxia-pretreated implants compared with the normoxia groups. VEGF level in plasma and peritoneal fluid were increased in hypoxia-pretreated group. However,the different value of TNFa between groups have on statistical significance.Conclusion(s): Hypoxia pretreatment play an important role in the originate and development of endometriosis by increase the proliferation and angiogenesis,decrease cellular apoptosis of implants in a rat model.