花生四烯酸细胞色素p450表氧化酶基因对野百合碱诱导的肺动脉高压大鼠IL-6,IL-8的影响

The effect of cytochrome P450 epoxygenase gene on expression of IL-6 and IL-8 in monocrotaline-induced pulmonary hypertension in rats

目的观察花生四烯酸色素p450表氧化酶基因CYP2J2过表达对野百合碱(MCT)诱导的肺动脉高压大鼠炎症细胞因子IL-6、IL-8、CRP和TNF-α表达的影响,探讨CYP2J2作用于肺动脉高压的机制。方法 :选取8周龄250-280gSD大鼠60只随机分为正常对照组(n=30)和模型组(MCT组)(n=30),模型组注射野百合碱(60mg/kg)造模。三周后分为6组:NS组(n=10),pCDNA3.1(n=10),pCDNA3.1-CYP2J2(n=10),MCT+NS(n=10),MCT+pCDNA3.1(n=10),MCT+pCDNA3.1-CYP2J2(n=10),注射质粒三周后,检测大鼠平均肺动脉压(mPAP),计算右心肥大指数(RVHl=RV/LV+S)。ELISA法检测大鼠血清中IL-6、IL-8、CRP和TNF-α水平。RT-PCR检测大鼠肺组织中IL-6和IL-8的mRNA表达。结果 :模型对照组mPAP、RVHI值、血清IL-6、IL-8、CRP和TNF-α水平及肺组织中IL-6和IL-8的mRNA表达均显著增高,与正常对照组相比有显著性差异(P<0.01);pCDNA3.1-CYP2J2治疗组mPAP、RVHI值、血清IL-6、IL-8、CRP和TNF-α水平及肺组织中IL-6和IL-8的mRNA表达均明显下调(P<0.05),但仍高于正常对照组(P<0.05)。结论花生四烯酸色素p450表氧化酶基因可通过抑制MCT诱导大鼠肺动脉高压模型中肺组织炎性细胞因子的表达、下调大鼠炎性细胞因子的分泌达到对肺动脉高压的治疗作用。

Objective To investigate the effect and mechanisms of Cytochrome P450 epoxygenase 2J2（CYP2J2）,a predominant human P450 arachidonic acid（AA） epoxygenase on the expression of inflammatory cytokines Ll-6,IL-8,CRP and TNF-α in monocrotaline（MCT） induced pulmonary hypertension in rats,and to investigate the therapeutic mechanism of CYP2J2 for pulmonary hypertension.Methods Sixty SD rats weighted 250～280g were allocated randomly into 2 groups,a normal control group and an MCT-induced pulmonary hypertension group（model control group）,were injected 60mg/kg MCT to induce pulmonary hypertension.Three weeks later,rats were divided into six groups：NS（n=10）,pCDNA3.1（n=10）,pCDNA3.1-CYP2J2（n=10）,MCT＋NS（n=10）,MCT＋pCDNA3.1（n=10）,MCT＋ pCDNA3.1-CYP2J2（n=10）.After 21 days,the mean pulmonary arterial pressure（mPAP） and right venirieular hypertophy index（RVHI） were measured,and the expression of IL-6 and IL-8 mRNA in the lung tissue and the levels of IL-6,IL-8,CRP and TNF-α in the serum were detected.Results The levels of mPAP,RVHI,content of plasma IL-6,IL-8,CRP and TNF-α,the mRNA expression of IL-6 and IL-8 in single MCT group were significantly increased compared with the normal group（P0.05）.The above parameters were significantly lower in the MCT＋ pcDNA3.1-CYP2J2 group than in the single MCT group（P0.05）,but remained higher than in the normal group（P0.05）.Conclusion CYP2J2 gene overpression has therapeutic effect on pulmonary hypertension induced by MCT through down-regulating IL-6,IL-8,CRP and TNF-α expression.