异甘草素通过调节Nrf2/HO-1氧化应激通路抑制ROS生成保护阿霉素诱导的药物性胰腺炎

Isoliquiritigenin ameliorates doxorubicin-induced acute pancreatitis by inhibiting ROS production via modulation of Nrf2/HO-1 oxidative stress pathway

背景多篇病例报道阿霉素(doxorubicin,DOX)引起急性胰腺炎的药物副作用,但尚无动物实验研究证实,且缺乏特异性的治疗药物.异甘草素(isoliquiritigenin,ISL)具有抗炎、抗氧化等多种药理活性,但是ISL对DOX引起药物性急性胰腺炎(drug-induced acute pancreatitis,DAP)的作用及机制尚不清楚.目的证实DOX可引起DAP的病理损伤,并阐明ISL保护DAP的作用及机制.方法将雄性25-30 g ICR小鼠随机分为对照组(Control组,腹腔注射等量生理盐水),DOX诱导的DAP模型组(DOX-DAP组,隔日腹腔注射DOX 10 mg/kg/只)和ISL治疗组(DOX+ISL组,隔日腹腔注射DOX 10 mg/kg/只,同时提前一天每日给予ISL灌胃100 mg/kg/只),每组8只.造模后5 d取材进行胰腺组织病理检测及评分;免疫组织化学方法检测胰腺组织alpha淀粉酶表达;免疫荧光法检测胰腺组织ROS生成;Western blot法检测胰腺组织Nrf2和HO-1蛋白表达.结果与Control组相比,DOX-DAP组小鼠胰腺组织呈现水肿、炎性细胞浸润等特征性病理损伤,胰腺组织病理学评分显著升高(P<0.001),胰腺组织alpha淀粉酶表达水平显著下降(P<0.01).与DOX-DAP组相比,ISL治疗组的胰腺组织病理评分显著降低(P<0.05),胰腺组织alpha淀粉酶表达水平显著升高(P<0.05).ROS荧光染色及Western blot检测显示,与Control组相比,DOX-DAP组小鼠胰腺组织ROS生成显著升高(P<0.001),Nrf2和HO-1蛋白表达水平轻度升高(P<0.001).与DOX-DAP组相比,ISL治疗组的胰腺组织ROS水平显著降低(P<0.001),Nrf2和HO-1蛋白表达水平显著升高(P<0.001).结论DOX能够引起小鼠胰腺出现以组织水肿和炎症细胞浸润为主要特征的病理损伤,ISL通过增强胰腺组织抗氧化应激水平对DOX引起的DAP具有保护作用,有望为DAP的预防与治疗提供新的方法.

BACKGROUND Several case studies have reported that doxorubicin(DOX)could induce acute pancreatitis,but no animal experiments have confirmed such side effect of DOX,and there is no specific treatment.Isoliquiritigenin(ISL)has a variety of pharmacological functions,including anti-inflammatory and antioxidant activities;however,the role and mechanism of ISL on DOX-induced acute pancreatitis(DAP)remain unclear.AIM To confirm whether doxorubicin(DOX)results in pancreatic tissue injury,and to determine the role and mechanism of isoliquiritigenin(ISL)in protecting against DOX-induced pancreatitis.METHODS Male ICR mice(25-30 g)were randomly divided into a control group(intraperitoneal injection of normal saline),DOX-DAP model group(intraperitoneal injection of DOX 10 mg/kg every other day),and ISL treatment group(DOX+ISL group;intraperitoneal injection of DOX 10 mg/kg every other day and intragastric administration of ISL 100 mg/kg per day),with 8 mice in each group.Pancreatic histopathology and scoring were performed 5 d after modeling.The expression of alpha amylase in pancreatic tissue was detected by immunohistochemistry.Immunofluorescence assay was used to detect ROS production in pancreatic tissue.Protein expression of Nrf2 and HO-1 in pancreatic tissue was detected by Western blot.RESULTS Compared with the control group,the mice in the DOX-DAP model group showed characteristic pathological damage,such as pancreatic tissue edema and inflammatory cells infiltration,with significantly increased histopathological scores(P<0.001)and decreased expression of alpha amylase in the pancreas(P<0.01).Compared with the DOX-DAP model group,the DOX+ISL group had significantly decreased histopathological scores(P<0.05)and increased expression of alpha amylase in the pancreas(P<0.05).ROS fluorescence staining and Western blot analysis showed that compared with the control group,ROS generation in pancreatic tissue in the DOX-DAP model group was significantly increased(P<0.001),and the expression levels of Nrf2 and HO-1 proteins were slightly increased(P<0.001).Compared with the DOXDAP group,the DOX+ISL group had significantly decreased ROS levels in pancreatic tissuesed(P<0.001),and significantly increased expression of Nrf2 and HO-1 proteins(P<0.001).CONCLUSION DOX can cause pancreatic pathological damage in mice,which is mainly characterized by pancreatic tissue edema and inflammatory cells infiltration.ISL administration has a protective effect on DOX-induced pancreatitis by enhancing the antioxidant stress level in pancreatic tissue,which is expected to provide a new method for the prevention and treatment of drug-induced pancreatitis.