Expression Characteristics And Significance Of NF-κB And INOS In Adrenal Disease

Objective To investigate the expressions of nuclear transcription facrorκB(NF-κB) and inducible nitric oxide synthase(iNOS) in adrenal disease,and to study the roles in the course of adrenal disease and clinical significance.Methods and results1:The acquisition of Adrenal cortical hyperplasia, cortical adenomas and normal adrenal tissue and the extraction of total RNAIn aseptic conditions, collect all the specimens in the freezing tube and stored at-196℃liquid nitrogen. A total of access to adrenal hyperplasia in 22 cases, adrenal cortical adenoma in 14 cases,12 cases of normal adrenal tissue.Isolate total RNA from tissues by Trizol instructions.Results:Case group specimens were confirmed by pathology. Specimens were obtained for total RNA each other..2.RT-PCR detection of NF-κB and iNOS gene expressionReverse transcription of cDNA from the total RNAof 48 specimens, and then take the cDNA products to PCR amplified, carried out by agarose gel electrophoresisResults:In the human adrenal cortex hyperplasia and adenoma tissue the expression of positive rates of NF-κB and iNOS mRNA were significantly higher than the normal group (P<0.05);There was no significant difference in the rate between the adrenal cortex hyperplasia and adenoma (P> 0.05),3.Quantitative PCR detection of NF-κB and iNOS gene expressionTake the total RNA samples of 48 cases to quantitative RT-PCR reaction product, using samples fitting analysis results,obtaine the threshold cycle number (Ct) values of NF-κB,iNOS andβ-actin.Results:The mRNA levels of NF-κB,iNOS in The human adrenal cortex hyperplasia and adenoma tissue were significantly higher than the normal group (P <0.05).There was no significant difference between the adrenal cortex hyperplasia and adenoma (P> 0.05),the expression of NF-κB,iNOS mRNA is positive Correlation(r=0.984 P<0.05)4.the acquisition of Immunohistochemical specimens and the NF-κB and iNOS protein expression in every groups.Select the conventional paraffin-embedded tissue blocks provided by the General Hospital of Guangzhou Military Region in July 2008-September 2009, Serial sections, film thickness Sμm, one case do HE staining, the remaining separate NF-κB, iNOS staining.Results:adrenal hyperplasia 13 cases, cortical adenomas 13 cases,15 cases of pheochromocytoma and 10 cases of normal adrenal tissue.the NF-κB and iNOS protein expression is positive or strong positive in all casese, the normal group did not express or weakly positive. The expression intensity of NF-κB and iNOS protein in the pheochromocytoma tissue was significantly higher than in the adrenal cortex hyperplasia and cortical adenomas(P<0.05).And the expression intensity of the latter was significantly higher than the normal group (P<0.05).Conclusion1.In human adrenal cortical hyperplasia, cortical adenoma and pheochromocytoma tissues, high expression of NF-κB mRNA and NF-κB protein were closely related to the occurrence and development;2.In human adrenal cortical hyperplasia, cortical adenoma and pheochromocytoma tissues, high expression of iNOS mRNA and iNOS protein were closely related to the occurrence and development;3.The expression of NF-κB and iNOS mRNA or protein are in the pathogenesis of interaction,mutual restraint, this may be the new targets of treatment and diagnosis of adrenal disease.