| ObjectiveThe purpose of this study was to investigate the expression of inducible nitric oxide synthase(iNOS) and Matrix Metalloproteinase-9(MMP-9) messenger ribonucleic acid (mRNA) in the endometrium of patients with Dysfunctional Uterine Bleeding(DUB), and to explore the roles of MMP-9 and iNOS in pathogenesis of uterine bleeding and the relations between them and dysfunctional uterine bleeding.Methods1. Tissue collectionAll endometrial tissue samples were randomly collected from women undergoing a gynaecological operation at the first clinical hospital China Medical University dating from Oct. 2005 to March 2006. Two groups of women participated in the study of endometrium: 1)study group:60 women, harf of all these tissue samples are simple hyperplasia, the other harf of them are complex hyperplasia; 2)control group: The control group including 20 samples which origin from uterine smooth muscle tumor patients whose menstrual cycle were normal and have had abdominal hysterectomy. All the women recruited to the study had not been exposed to steroid for three months prior to operation, did not have either an intrauterine contraceptive device(IUD)in situ. All the tissue samples were conserved in liquid nitrogen soon after operation(within 5 minutes)for 4 hours then transported to -70℃deep freeze refrigerator to be conserved.2. MethodsReverse transcriptase polymerase chain reaction ( RT-PCR) was used to study the expression of iNOS and MMP-9mRNA.(1)Total RNA was extracted with RNAout Reagent, total RNA was prepared by solvent extraction following the instructions.(2)All procedures of RT and PCR ractions were implemented according to the illustration of RNA PCR Kit. Reactions of RT were incubated at 30℃for 10 min followed by 30 min at 42℃and 5 min at 99℃. The amplification profile was as follows: 94℃for 2 min; 94℃for 30 sec, 57℃for 40 sec and 72℃for 1 min; 35cycles and 72℃for 10min.(3) Statistical Analysis: The data were evaluated by independent-samples T test of SPSS 13. 0.Results1. The expression of iNOS and MMP-9 mRNA have small amounts in normal endometrium.2. The expression of MMP-9 mRNA was higher in the endometrium of simple hyperplasia and complex hyperplasis than those in normal endometrium, there is significant difference between them (P<0. 01).3. The expression of iNOSmRNA was higher in the endometrium of simple hyperplasia and complex hyperplasis than those in normal endometrium, there is significant difference between them (P<0. 01).4. The expression of MMP-9 mRNA was higher in the endometrium of complex hyperplasis than those of simple hyperplasia, there was significant difference between two groups of different pathological types in the study group of endometrium(P<0. 01).5. The expression of iNOSmRNA was higher in the endometrium of complex hyperplasis than those of simple hyperplasia, there was significant difference between two groups of different pathological types in the study group of endometrium(P<0. 01).Conclusions1.The higher expression of iNOS and MMP-9 lead to abnormal bleeding of uterine, iNOS and MMP-9 are important regulating factors in pathogenesis of dysfunctional uterine bleeding.2. There is closed relation between the expression of iNOS ,MMP-9 and pathological types of endometrium, this suggest that iNOS and MMP-9 are involved in pathogenesis of dysfunctional uterine bleeding. |
