apocynin对急性坏死性胰腺炎大鼠肺组织的保护作用

Protective effect of apocynin on lung injury induced by acute necrotic pancreatitis in rat

目的探讨还原型辅酶Ⅱ氧化酶(NOX2)抑制剂apocynin对急性坏死性胰腺炎(ANP)模型大鼠肺损伤的保护作用。方法 40只SPF级Wistar大鼠采用完全随机法随机分为4组:假手术组(SO组,10只)、ANP模型组(ANP组,12只)、apocynin治疗组(APO组,10只)及APO药物对照组(APO-CON组,8只)。采用胆胰管逆行注射5%牛磺胆酸钠溶液制备ANP模型,造模前30 min时APO组注射APO干预,APO-CON组和SO组仅开腹翻动胰腺和十二指肠同时在与APO组相同时间点分别注射apocynin和10%二甲基亚砜干预。各组手术后12 h经大鼠下腔静脉取血并处死大鼠后取肺组织,检测大鼠血清淀粉酶和脂肪酶水平,采用免疫组织化学方法检测肺组织中核因子-κB(NF-κB)、肿瘤坏死因子(TNF)-α和NOX2蛋白表达,用免疫荧光法检测肺组织中髓过氧化物酶(MPO)、Toll样受体4(TLR4)和CD68蛋白表达,采用ELISA法检测丙二醛(MDA)浓度及超氧化物歧化酶(SOD)活性。结果与SO组比较,ANP组大鼠血清淀粉酶和脂肪酶水平以及肺组织中NF-κB、TNF-α、NOX2、MPO、TLR4、CD68蛋白表达及MDA浓度均明显升高(P<0.05),与ANP组比较,APO组的上述指标均明显降低(P<0.05);SOD水平在ANP组明显低于SO组(P<0.05),而其在APO组明显高于ANP组(P<0.05)。结论 apocynin可能通过抑制NOX2活性改善大鼠ANP症状并减轻肺损伤。

Objective To investigate protective effect of apocynin, the inhibitor of NADPH oxidase Ⅱ（NOX2）,on lung injury induced by acute necrotic pancreatitis（ANP） in rat. Methods Forty SPF adult male Wistar rats were randomly divided into 4 groups： shame operation group（SO group, n=10）, ANP model group（ANP group, n=12）,apocynin treated group（APO group, n=10）, and apocynin control group（APO-CON group, n=8）. The ANP models were induced by the retrograde injection of 5% sodium taurocholate through the biliopancreatic duct in the ANP group and the APO group. The apocynin was injected at 30 min before the induction of ANP models in the APO group. The pancreas and duodenum of rats were just flipped and the apocynin and the 10% DMSO（2 mL/kg） were injected in the APO-CON group and SO group respectively. All the rats were sacrificed at 12 h after the operation. The blood samples were collected by the inferior vena cava puncture, and the levels of serum amylase and lipase were measured by the auto-chemistry analyzer. The lung tissues were harvested and the integrated optical densities（IODs） of the nuclear factor-κB（NF-κB）,tumor necrosis factor-α（TNF-α）, and NOX2 were detected by the immunohistochemistry assay. The IODs of the myeloperoxidase（MPO）, Toll like receptor 4（TLR4）, and CD68 were detected by the immunofluorescence assay. The concentration of malondialdehyde（MDA） and activity of superoxide dismutase（SOD） were tested by the ELISA method.Results The levels of the serum amylase and lipase and the IODs of the NF-κB, TNF-α, NOX2, MPO, TLR4, CD68, and concentration of MDA of the lung tissues in the ANP group were significantly increased as compared with the SO group（P〈0.05）, these indices in the APO group were significantly decreased as compared with the ANP group（P〈0.05）. The SOD activity of the lung tissue in the ANP group was significantly decreased as compared with the SO group（P〈0.05）,which in the APO group was significantly increased as compared with the ANP group（P〈0.05）. Conclusion Apocynin can ameliorate lung injury induced by ANP through inhibiting activity of NOX2.