Wang Fang, Liu Jian, Ye Yingfa, Zhang Xiaojun, Wan Lei
Anhui University of Traditional Chinese Medicine, Hefei 230031, China.
First Affiliated Hospital of Anhui University of Traditional Chinese Medicine, Hefei 230031, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2014 May;30(5):497-501.
To investigate the mechanism underlying the changes in the cardiac function in rat models of Sjogren's syndrome (SS) based on Kelch-like ECH-associated protein 1 (Keap1)-Nrf2/ARE signaling pathways.
Thirty male Wistar rats were randomly divided into two groups, normal control (NC) group and SS model group, with 15 in each. The rats in the model group were injected with the complete Freund's adjuvant plus homologous antigen of submandibular gland (0.2 mL mixture) into bilateral posterior paw metatarsus. The body mass, water intake, submandibular gland index, spleen index, and histological changes of the glands were observed 30 days after inflammation was induced. Cardiac function was assessed using invasive hemodynamic monitoring. Meanwhile, reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD) and total antioxidant capacity (TAC), IL-18, IL-35 were detected using ELISA; ROS and reactive nitrogen species (RNS), glutathione (GSH), thioredoxin (TrX) protein were determined using immunohistochemical staining. The expressions of Keap 1, nuclear factor erythroid 2-related factor (Nrf2), macrophage activating factor (Maf) antioxidant responsive element (ARE) mRNAs were detected using real-time fluorescent quantitative PCR (qRT-PCR); the levels of heme oxygenase-1 (HO-1), γ-glutamic acid and a half long glycine synthetase (γ-GCS) proteins in cardiac tissue were examined using Western blotting.
Compared with the NC group, the model group presented reduced body mass, submandibular gland index and spleen index (P<0.05), increased water intake (P<0.01), heart rate (HR), heart index (HI), left ventricular systolic pressure (LVSP) and left ventricular diastolic pressure (LVEDP) (P<0.05), and decreased left ventricular ±dp/dtmax (P<0.05). Compared with the NC group, the model group had increased IL-18, MDA, RNS, TAC, ROS levels, and Keap1, Maf, Nfr2 mRNAs, HO-1, γ-GCS protein expressions in the heart tissue, while TrX, GSH, IL-5 and SOD levels decreased significantly (P<0.01).
The immune imbalance in SS rats may be related with the up-regulated levels of Keap1, Maf, Nfr2 mRNAs, HO-1, γ-GCS.
基于类 Kelch 样 ECH 相关蛋白 1（Keap1）-核因子 E2 相关因子 2（Nrf2）/抗氧化反应元件（ARE）信号通路，探讨干燥综合征（SS）大鼠模型心功能改变的机制。
将 30 只雄性 Wistar 大鼠随机分为两组，正常对照组（NC 组）和 SS 模型组，每组 15 只。模型组大鼠双侧后足跖注射完全弗氏佐剂加下颌下腺同源抗原（混合物 0.2 mL）。诱导炎症 30 天后，观察大鼠体重、饮水量、下颌下腺指数、脾脏指数及腺体组织学变化。采用有创血流动力学监测评估心功能。同时，用酶联免疫吸附测定法（ELISA）检测活性氧（ROS）、丙二醛（MDA）、超氧化物歧化酶（SOD）和总抗氧化能力（TAC）、白细胞介素 18（IL-18）、白细胞介素 35（IL-35）；用免疫组织化学染色法测定 ROS 和活性氮（RNS）、谷胱甘肽（GSH）、硫氧还蛋白（TrX）蛋白。用实时荧光定量聚合酶链反应（qRT-PCR）检测 Keap1、核因子红细胞 2 相关因子（Nrf2）、巨噬细胞激活因子（Maf）、抗氧化反应元件（ARE）mRNA 的表达；用蛋白质印迹法检测心脏组织中血红素加氧酶-1（HO-1）、γ-谷氨酸半胱氨酸合成酶（γ-GCS）蛋白水平。
与 NC 组比较，模型组大鼠体重、下颌下腺指数和脾脏指数降低（P<0.05），饮水量增加（P<0.01），心率（HR）、心脏指数（HI）、左心室收缩压（LVSP）和左心室舒张压（LVEDP）升高（P<0.05），左心室±dp/dtmax 降低（P<0.05）。与 NC 组比较，模型组心脏组织中 IL-18、MDA、RNS、TAC、ROS 水平升高，Keap1、Maf、Nfr2 mRNA、HO-1、γ-GCS 蛋白表达增加，而 TrX、GSH 的水平及 IL-5 和 SOD 水平显著降低（P<0.01）。
SS 大鼠免疫失衡可能与 Keap1、Maf、Nfr2 mRNA、HO-1、γ-GCS 水平上调有关。
