| Obsjective:To investigate the expression levels of endothelial and inducible nitric oxide synthase(eNOS and iNOS) in eutopic and ectopic endometrial of women with endometriosisand normal endometrial tissues throughout the menstrual cycle from molecular level and its efect in the mechanism of endometriosis. This experimentcan be chosen for endometriosis to targeted therapy in clinic.Material and MethodsThis was a prospective randomized study undertaken in Women's Hospital School of Medicine Nanchang University from October 2006 to May 2007.Thirty-five patients were divided into two groups:20 patients in Control group and 15 patients in Experiment group.Experiment group were divided into two groups:Eutopic endometrial group and Ectopic endometrial group.Experiment group were 15 patients with ovarian cyst of endometriosis diagnosed by laparoscopic surgery,including 15 ovarian endometriosis and 15 eutopic endometrium (proliferative stage 18,progestational stage12) ;Control groupwere 20 patients with disease of cervix by diagnostic curettage, including20 ornormal uterine endometrium (proliferative stage 15, progestational stage5). All the patient were ruled out other disease of endometrium.The mean age of experiment group was32.2±2.77,the mean age of control group was34.9±3.24,No statistically significant differences of age level was found between experiment group and control group.All the patient must have potentia generandi,age less then 45,no hypohepatia,no nephropathy , no blood disease,and no inflammation or anoxic disease;had not sex hormone and immunosuppressant to cure before six months.The endometriosis were rapidly removed and washed in ice-cold 0.9% NaCL solution, then put it into EPcuvette and let it in lox for halfhour transfer it into-70℃icebox for use. To measure transcription of eNOS and iNOS calcium channel,semiquantitative RT-PCR was performed. Total RNA was extracted with TRNZOL Reagent.The quality of RNA content was assessed by Electrophoretic diagram by agarose gel . A quantity of 5μg of total RNA was reversetranscribed in a 25μl reaction mixture .For PCR 2ul of the cDNA mixture was added to 25 ul of a master mix.PCR products were separated on 1.5% agarose ethidiutm-bromide gel, visualized under ultraviolet ligh. By reverse transcription-polymerase chain reaction ,we assess the expression levels of endothelial and inducible nitric oxide synthase(eNOS and iNOS) in eutopic and ectopic endometrial of women with endometriosisand normal endometrial tissues throughout the menstrual cycle from molecular level.We observed the diference betweenthe expression levels of NOS and iNOS inthe ectopic endometriosis and eutopic endometriosis.All the date were dealed with statistical software SPSS 13.0.Results:1.In all endometriotic samples of proliferative phases,the expression of eNOS mRNA level was found Expression of eNOS mRNAs was significantly increased in Eutopic endometrial group and Ectopic endometrial group contrast with control group (P=0.0001)But No changes in mRNAs level of eNOS were observed between Eutopic endometrial group and Ectopic endometrial group (p=0.759)2.In all endometriotic samples of secretory phases,the expression of eNOS mRNA level was found Expression of eNOS mRNAs was significantly increased in Eutopic endometrial group contrast with control group (P=0.049)Also as the Ectopic endometrial group(P=0.038);But No changes in mRNAs level of eNOS were observed between Eutopic endometrial group and Ectopic endometrial group (p=0.793)3.In proliferative phases,these is not expression level of inducible nitric oxide synthase mRNA in experiment endometrium and normal uterine endometrium.4.In all endometriotic samples of secretory phases,the expression of iNOS mRNA level was found.Expression of iNOS mRNAs was significantly increased in Eutopic endometrial group contrast with control group(P=0.05)Also as the Ectopic endometrial group(P=0.012);No statistically significant diferences of iNOSmRNA level was found between Eutopic endometrial group andEctopic endometrial group(P=0.822).5.In Eutopic endometrial group, eNOS mRNA level was higher in the proliferative phases than in secretory phase(p=0.001),inEctopic endometrial group, eNOS mRNA level was higher in the proliferative phases than in secretory phase(p=0.004)but no significantly higher (P=0.072)between the proliferative phases and in secretory phase.Conclusion:1.Greater amounts of NOS are present in the endometrial tissues of women with endometriosis,implying a possible role for NO in the pathogenesis of endometriosis2.No statistically significant diferences of iNOS and eNOS mRNA level was found between Eutopic endometrial group and Ectopic endometrial group... |
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