山东医药

目的 基于PTEN诱导假定激酶（PINK1）/E3泛素连接酶（Parkin）介导线粒体自噬探讨核因子E2相关因子2(Nrf2)在急性胰腺炎（AP）氧化应激及炎症调节中的作用机制。方法 将SPF级雄性SD大鼠随机分为5组，每组10只。Sham组开腹翻动肠道后关腹，AP组开腹后经胰管逆行入路注射3.5%牛磺胆酸钠建立AP模型后关腹，TBHQ组建立AP模型关腹后腹腔注射Nrf2激活剂TBHQ,ML385组建立AP模型关腹后腹腔注射Nrf2抑制剂ML385,TBHQ+Mdivi-1组建立AP模型关腹后腹腔注射Nrf2激活剂TBHQ和线粒体自噬抑制剂Mdivi-1。采用ELISA法检测大鼠血清淀粉酶（AMS）、脂肪酶（LPS）、肿瘤坏死因子α(TNF-α)及白细胞介素6(IL-6)水平；HE染色观察大鼠胰腺组织病理学变化；DCFH-DA荧光探针检测胰腺组织活性氧（ROS）水平；RT-qPCR及Western blotting分别检测胰腺组织Nrf2、PINK1、Parkin、p62、LC3B mRNA及蛋白表达。结果 与Sham组相比，AP组可见胰腺组织水肿及炎症损伤，AMS、LPS、TNF-α、IL-6及ROS升高，Nrf2、PINK1、Parkin及LC3B表达降低，p62表达升高（P均<0.05）。与AP组相比，TBHQ组胰腺病理损伤减轻，AMS、TNF-α、IL-6及ROS降低，Nrf2、PINK1、Parkin及LC3B表达升高，p62表达降低（P均<0.05）;ML385组病理损伤加重，AMS、LPS、TNF-α、IL-6及ROS升高，Nrf2、PINK1、Parkin及LC3B表达降低，p62表达升高（P均<0.05）。与TBHQ组相比，TBHQ+Mdivi-1组病理损伤进一步加重，AMS、LPS、TNF-α、IL-6及ROS升高，PINK1、Parkin及LC3B表达降低，p62表达升高（P均<0.05）。结论 Nrf2激活可减轻AP氧化应激及炎症反应，其机制可能与Nrf2促进PINK1/Parkin介导线粒体自噬有关。

Objective To investigate the regulatory effect of nuclear factor erythroid 2-related factor 2(Nrf2) on PTEN-induced kinase 1(PINK1)/Parkin-mediated mitophagy and to explore its underlying mechanism in oxidative stress and inflammatory responses in acute pancreatitis(AP).Methods Fifty SPF male SD rats were randomly divided into five groups, with 10 rats in each group: the Sham group(received laparotomy with intestinal manipulation followed by abdominal closure), the AP group(underwent laparotomy, followed by retrograde injection of 3.5% sodium taurocholate through the pancreatic duct to establish an AP model, after which the abdomen was closed), the TBHQ group(received intraperitoneal injection of the Nrf2 activator TBHQ after AP model establishment and abdominal closure), the ML385 group(received intraperitoneal injection of the Nrf2 inhibitor ML385 after AP model establishment and abdominal closure), and the TBHQ + Mdivi-1 group(received intraperitoneal injection of both the Nrf2 activator TBHQ and the mitophagy inhibitor Mdivi-1 after AP model establishment and abdominal closure), respectively.Serum levels of amylase(AMS), lipase(LPS), tumor necrosis factor-α(TNF-α), and interleukin-6(IL-6) were measured by enzyme-linked immunosorbent assay(ELISA).Histopathological changes in pancreatic tissues were evaluated by HE staining.Reactive oxygen species(ROS) levels in pancreatic tissues were detected using the DCFH-DA fluorescent probe.The mRNA and protein expression levels of Nrf2, PINK1, Parkin, p62, and LC3B in the pancreatic tissues were determined by RT-qPCR and Western blotting, respectively.Results Compared with the Sham group, the AP group exhibited pancreatic edema and inflammatory injury, accompanied by significantly increased levels of AMS, LPS, TNF-α, IL-6, and ROS, decreased expression of Nrf2, PINK1, Parkin, and LC3B, and increased p62 expression(all P<0.05).Compared with the AP group, the TBHQ group showed attenuated pancreatic pathological injury, reduced levels of AMS, TNF-α, IL-6, and ROS, increased expression of Nrf2, PINK1, Parkin, and LC3B, and decreased p62 expression(all P<0.05).In contrast, the ML385 group exhibited aggravated pancreatic injury, increased levels of AMS, LPS, TNF-α, IL-6, and ROS, decreased expression of Nrf2, PINK1, Parkin, and LC3B, and increased p62 expression(all P<0.05).Compared with the TBHQ group, the TBHQ + Mdivi-1 group exhibited more severe pancreatic injury, accompanied by increased levels of AMS, LPS, TNF-α, IL-6, and ROS, decreased expression of PINK1, Parkin, and LC3B, and increased p62 expression(all P<0.05).Conclusion Activation of Nrf2 attenuates oxidative stress and inflammatory response in AP, and the mechanism may be related to Nrf2 promoting PINK1/Parkin-mediated mitophagy.