NADPH氧化酶抑制剂Apocynin预防脂多糖心肌纤维化的研究

摘要

Objective To investigate the preventive effects of apocynin,an NADPH oxidase inhibitor,on cardiac fibrosis in mice exposed to subclinical lipopolysaccharide.Methods Totally 40 male C57BL/6J mice were divided randomly into 4 groups:control group,apocynin group,LPS group and LPS + apocynin group.LPS was injected intraperitoneally once a week for 4 weeks to mimic cardiac fibrosis in LPS group and LPS + apocynin group.Apocynin was given in apocynin group and LPS + apocynin group,once a week for 4 weeks.Collagen fraction area of the left ventricle was measured with picrosirius red staining.The cardiac expression of fibrosis-related genes was measured by RT-PCR.The subunits gp91 phox and p67phox of NADPH oxidase were analyzed by Western blot,ROS production was determined by DHE staining.Results The cardiac fibrosis of mice occurred after 4 weeks.Compared with control group,the interstitial diffuse fibrous tissue in myocardium,the collagen volume fraction,the mRNA expression of LPS-induced activation of fibrosis-related genes Col Ⅰ a 1,Col Ⅲ a1,MMP2 and MMP9,the expression gp91 phox and p67phox and ROS production in LPS group were increased.Compared with LPS group,apocynin decreased the above indexes significantly (P ＜ 0.05).Conclusion Apocynin can protect against the cardiac fibrosis in mice exposed to subclinical LPS by inhibiting the over-expression of myocardial gp91phox and p67phox.%目的 探讨NADPH氧化酶抑制剂Apocynin对脂多糖(Lipopolysaccharide,LPS)心肌纤维化的预防作用.方法 40只小鼠随机分成4组:对照组(CON)、Apocynin组、LPS组和LPS+ Apocynin组,每组10只.LPS组和LPS+ Apocynin组给予LPS 10 mg/kg,每周1次,连续4周;Apocynin组和LPS+ Apocynin组给予Apocynin 10 mg/kg,1次/d,连续4周.采用天狼星红染色评估心肌纤维化程度,RT-PCR检测Col Ⅰ al、ColⅢal、MMP2、MMP9 mRNA的表达,Western blot检测心肌gp91 phox和p67 phox的表达,DHE检测心肌活性氧生成.结果 小鼠LPS注射4周后出现心肌纤维化.与对照组比较,LPS组心肌间质内弥漫性纤维组织显著增多,胶原容积分数增加,心肌组织中胶原相关因子Col Ⅰ a1、ColⅢa1、MMP2、MMP9 mRNA表达明显增加,心肌gp91 phox和p67 phox的表达增加,ROS生成增加(P＜0.05).与LPS组比较,Apocynin干预可抑制上述指标的增加(P＜0.05).结论 Apocynin能预防LPS诱导小鼠心肌纤维化,其效应可能与抑制心肌gp91phox和p67phox过表达有关.