The Effect On Blood Pressure Regulation Of Cytochrome P450 4A11

Cytochrome P450 4s(CYP4s)are expressed in many organs of the human body and can metabolize AA to 20-HETE by hydroxylation.Expression or activity changes of CYP4 s can cause changes in the amount of 20-HETE.20-HETE was reported to be acted in different parts of the blood pressure regulation.On the one hand,20-HETE can be acted as a very effective vasoconstrictor;on the other hand,it can promote urinary sodium excretion in the proximal part of renal tubule and medullary loop,and inhibit sodium reabsorption to anti-hypertension.Whether CYP4 s effectively regulate blood pressure at different sites through 20-HETE is not clear.In order to further investigate the relationship between CYP4 s and blood pressure regulation and whether it was achieved by 20-HETE.For the more possible CYP4 s enzyme related to essential hypertension(EH),at first,it conducted a study on the relationship between EH and SNPs of CYP4 s family and related ubiquitination including CYP4A11,CYP4F12,CYP4F2,CYP4F3,CUL3 and KLHL3.It was found that CYP4A11,CYP4F3,CUL3 and KLHL3 SNPs had been significant relationship to EH,and CYP4A11 enzyme could significantly increase the risk of EH,so it was selected as target enzyme into further investigation by animals,organs and cells tests to verify the regulation effect of CYP4A11 on blood pressure,and whether through 20-HETE.First of all,the lentiviral vector expressing the Cyp4a10 with homologue and similar function of CYP4A11 in mice with green fluorescence was constructed,sequenced and compared and it was transfected in two cell lines to verify the integrity and availability of the vector.Then,the lentiviral vector was stably transfected into the homologous C57BL/6J and e NOS(-/-)male mice to establish stable overexpression of Cyp4a10 mice models by tail vein injection,and the transfection effect was evaluated by fluorescence coloration,immunocytochemistry,protein expression and m RNA levels detection in multiple organs,and the results demonstrated that Cyp4a10 overexpression into the mice models.The mice were monitored systolic blood pressure,and it were investigated the effect between Cyp4a10 expression and blood pressure,whether it was affected by HET0016(20-HETE synthesis and Cyp4 a enzyme specific inhibitor),and the changes of 20-HETE in mice body fluid.The results showed that the expression of Cyp4a10 is related to blood pressure,and can be blocked by HET0016,the expression of Cyp4a10 is positively correlated with the 20-HETE in the body fluid of mice,so it was speculated that CYP4A11 may promote mice increased blood pressure,and 20-HETE play an effective role.Secondly,the effect of Cyp4a10 on vascular tension and the possible relationship with 20-HETE were observed in the main and renal arteries of above-mentioned mice in vitro.The results showed that the aorta of mice overexpressed Cyp4a10 was significantly less elastic than that of wild-type mice after adrenaline-induced contraction after endothelial function was limited by Nitro-L-arginine methyl ester(L-NAME).And after of overexpression of Cyp4a10,protein and m RNA levels of angiotensin converting enzyme-1(ACE1),angiotensin II receptor-1(AT1R),endothelin-1(ET1),endothelin-1 receptor(ETAR),endothelial nitric oxide synthase(e NOS)such vascular tension factors and adenosine phosphate enzyme alpha(ATPase ?)in the kidney were investigated in aorta and kidney of mice models,the results found in isolated organs was significantly upregulated in the contraction factors and reduced diastolic factor,and it can be speculated that homology and similiar function CYP4A11 could significantly change the tension performance of the aorta and renal artery of mice,and the effects on vascular regulatory factors is a vasoconstrictor.Thirdly,in the cell tests,obtaining CYP4A11 plasmid and designing and screening si RNAs to silence CYP4A11 expression,and then transfected into cell lines to construct 4 cell models.Then examine the CYP4A11 enzyme in human EA.hy926 and HK-2 cell lines in expression or cell lines after silencing the multiple vascular regulation factors,and the results showed that CYP4A11 was positively correlated with protein and m RNA expression of above vascular regulation factors,and confirmed that 20-HETE played a role in the effects.Through the tests of mice,isolated organs and cell lines,the study subject was found that CYP4A11 enzyme can significantly promote the regulation of blood pressure,when the expression is increased,promoting contraction and blood pressure increasement and the regulation of blood vessels likely to through 20-HETE.It may be the mechanism of CYP4A11 SNP is related to EH obviously.This project was a basis to improve the mechanism of CYP4A11 regulating blood pressure,and provides new ideas and new strategies for the diagnosis,treatment and prevention of EH.