The Mechanism Research Of EETs Metabolized By CYP450of Arachidonic Acid In Severe Hepatitis B

Background and objective High incidence of severe hepatitis, hazard, verycomplex pathogenesis, clinical lack of specific effective treatment. Studies EETshave to dilate blood vessels, anti-inflammatory, promoting the formation of new bloodvessels, promote endothelial cell proliferation, migration, inhibition of endothelial cellapoptosis, promote tumor proliferation and metastasis, and so on. The epoxygenaseCYP enzyme（sCYP450）generate epoxyeicosatrienoic acids (EETs), by catalysing theepoxidation of arachidonic acid olefin bonds, resulting in the production of fourregioisomeric EETs:5,6-EETs,8,9-EETs,11,12-EETs and14,15-EETs. The EETs aremetabolized to their di-hydroxyl derivatives (DHET) by the soluble epoxide hydrolase(sEH) Recently, sEH inhibitors (sEHIs) have been developed to enhance the actions ofEETs. CYP450is Ι metabolic enzymes, catalyzed many xenobiotics metabolism andsteroid hormone, arachidonic acid, etc. The synthesis and metabolism of endogenoussubstances. CYP2C9is one of the family of CYP2C, isozyme, mainly distributed inliver tissue, and accounts for about20%of the total liver microsomal CYP enzyme.The rapid development of sEHIs for in vivo use and clinical testing in the past decadeis remarkable. A landmark study showed that injection of a sEHI to the spontaneouslyhypertensive rat (SHR) and angiotensin-induced hypertension lowered blood pressure.Another breakthrough showed that an orally administered sEHI was antihypertensive and slowed the progression of renal damage. Following this, a number of studies haveprovided exciting findings on the broad potential for sEHIs as cardiovasculartherapeutic agents, and a first in class sEHI began clinical Phase IIa testing this year.This research through the MHV-3infection to establish animal model of viralfulminant hepatitis in mice study arachidonic acid oxide metabolites EETs CYP450table in the role of viral fulminant hepatitis and its possible mechanism. And thesurvival time of mice will extend or not when add the sEH inhibitors of MHV-3virusinfection,,whether the sEHI can be the new severe hepatitis therapeutic agents.Methods Establish Balb/c mice MHV-3virus infection model with ELISAmethod to detect different time points within the liver, serum11,12-EETs,14,15-EETs content changes; Using immunohistochemistry, western blot and other methodsto detect whether EETs in the liver by arachidonic acid by CYP2C9. Add with sEHinhibitors AUDA5mg/kg slows the degradation of EETs observe the survival time ofmice after the presence of extensionResults1, In the course of fulminant hepatitis,11,12-EETs and14,15-EETsincreased, at the time of72h (P <0.05, with statistical significance). Prompt EETs inMHV-3induced pathological process of fulminant hepatitis in mice.2, In MHV-3fulminant hepatitis virus infection mice liver tissue slices, rows of CYP2C9immunohistochemical not see positive results; In MHV-3fulminant hepatitis virusinfection mice liver tissue protein extraction, do western blot to see positive results.Show the EETs in the liver tissue is not produced by arachidonic acid via enzymeCYP2C9.3, add with sEH inhibitors of MHV-3virus infection after AUDA,11,12-EETs gradually increased, at the time of72h (P>0.05, without statisticalsignificance). But it does not extend survival time of mice.Conclusion1.EETs participation MHV-3-induced murine viral fulminant hepatitispathological process, but did not block the progress of fulminant hepatitis, can notimprove the survival time of mice of fulminant viral hepatitis. 2.sEH inhibitor AUDA failed to mice violence liver model liver11,12-EETs increasedsignificantly, no improvement in the survival rate of mice; In this model, liverCYP2C9expression, suggesting that in this model, possible liver CYP2C9damaged,leading to a reduction in upstream EETs generated, failed to play the role ofanti-inflammatory protection.11,12-EETs,14,15-EETs increase may be related toother types of table oxidase.