The Relation Between INOSmRNA In Endometrium And Anovulatory Functional Bleeding

ObjectiveThe purpose of this study was to investigate the expression and distribution patterns of inducible nitric oxide synthase messenger ribonucleic acid ( iN-OSmRNA) in the endometrium of patients with anovulatory functional bleeding, and to explore its role in pathogenesis of uterine bleeding , and the relation between it and anovulatory functional bleeding.Method1. 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. 2004 to June 2005. Two groups of women participated in the study of endometrium: 1) study group:60 women,of all these tissue samples 40 of them are simple hyperplasia ,20 of them are complex hyperplasia;2) control group:30 women. All the women of control groups participated in the study were adults with primitively normal mestrual cycles. 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. Each of the specimens was separated into two smaller specimens for both immunohistochem-istry and RT - PCR.2. MethodsRT-PCR:RT - PCR was used to study the expression of iNOSmRNA. (l)Total RNA was extracted with RNAout Reagent, total RNA was prepared by solvent extraction following the instructions. (￡)A11 procedures of RT and PCR raction were implemented according to the illustration of RNA PCR Kit. Reactions of RT were incubated at 30T! for 10 min followed by 30 min at 42T1 and 5 min at 99^1. The amplification profile was as follows: 94°C for 2 min;94X1 for 30 sec, 55°C for 30 sec and 72X, for 1 min;30 cycles and 72X, for 7 min.Immunohistochemistry (SP method)Immunohistochemistry was used to study the expression and distribution patterns of iNOS protein. All procedures were implemented according to the product illustration .3. Statistical Analysis;The data were evaluated by independent -samples T test.SSPS 13. 0.Results1. iNOS was detected in the glandular epithelial and some of the interstitial cells.2. The expression of iNOSmRNA, iNOS protein have small amounts in normal endometrium.3. The expression of iNOSmRNA, iNOS protein was higher in the endometrium of simple hyperplasia and complex hyperplasis than those in normal endometrium, there is significant difference between them (P <0. 05).4. The expression of iNOSmRNA, iNOS protein was higher in the endometrium of complex hyperplasis than those simple hyperplasia, there was significant difference between two groups of different pathological types in the study group of endometrium (P < 0.05).Conclusions1. The higher expression of iNOS lead to abnormal bleeding of uterine, andiNOS is an important regulating factor in pathogenesis of anovulatory functional bleeding.2. There is closed relation between the expression of iNOS and pathological type of endometrium, this suggest that iNOS is involved in pathogenesis of anovulatory functional bleeding.