文献类型：期刊文献

中文题名：基于Keap1-Nrf2信号通路探讨大补肺汤预防给药对高原低氧大鼠脑损伤的干预作用

英文题名：Intervention Effect of Dabufei Decoction(大补肺汤)on Acute Brain Injury in Rats with High-Altitude Hypoxia Based on Keap1-Nrf2 Signaling Pathway

作者：黄勇[1,2,3];李从艺[1,2,3];苏韫[1,2,3];龚红霞[1,2,3];曾元丁[1,2,3];冷光现[4];李莉霞[5];曹旺杰[1,2,3]

第一作者：黄勇

机构：[1]甘肃中医药大学基础医学院,兰州730000;[2]甘肃省高校重大疾病分子医学与中医药防治研究省级重点实验室,兰州730000;[3]敦煌医学与转化教育部重点实验室,兰州1730000;[4]兰州大学第二医院,兰州730000;[5]天水市中西医结合医院,天水741000

第一机构：甘肃中医药大学基础医学院（敦煌医学研究所）

年份：2024

卷号：40

期号：7

起止页码：27

中文期刊名：中药药理与临床

外文期刊名：Pharmacology and Clinics of Chinese Materia Medica

收录：;北大核心:【北大核心2023】；CSCD:【CSCD2023_2024】；

基金：国家自然科学基金项目(编号:82060833);甘肃省中医药管理局项目(编号:GSKP-2021-32);敦煌医学与转化教育部重点实验室开放课题项目(DHYX22-04);甘肃省高等学校青年博士基金(编号:2022QB-102)。

语种：中文

中文关键词：大补肺汤;高原低氧;脑损伤;Kelch样环氧氯丙烷相关蛋白-1/核因子E2相关因子2信号通路

外文关键词：Dabufei decoction(大补肺汤);High-altitude hypoxia;Brain injury;Keapl/Nrf2 signaling pathway

摘要：目的:探讨预防给药大补肺汤通过调控Kelch样环氧氯丙烷相关蛋白-1(Keap1)/核因子E2相关因子2(Nrf2)信号通路及相关分子的表达,从而对高原低氧大鼠脑损伤的干预作用。方法:60只SPF级SD大鼠随机分为正常对照组、模型对照组、地塞米松0.5 mg/kg组、大补肺汤8.8、17.6、35.1 g/kg组,大补肺汤各组灌胃给予相应药物,连续给药14 d,地塞米松组腹腔注射给药,进舱前连续给药3 d,其余组给予等体积生理盐水。除正常对照组外,第15 d起各组大鼠于实验动物低压模拟舱中进行低氧暴露,连续3 d。造模结束后处死动物,HE染色观察大鼠脑组织海马形态;TBA法、比色法、微板法分别检测脑组织中丙二醛(MDA)含量、谷胱甘肽过氧化物酶(GSH-Px)活性、谷胱甘肽(GSH)的含量;Werstern blot和RT-qPCR法检测大鼠脑组织中Keap1、Nrf2、血红素加氧酶1(HO-1)、醌氧化还原酶-1(NQO-1)蛋白与mRNA的表达。结果:HE结果显示模型对照组大鼠海马CA1区锥体细胞水肿,排列不规则,胞质疏松淡染;与正常对照组比较,GSH-Px活性和GSH含量明显降低(P<0.05或P<0.01),MDA含量显著升高(P<0.01),Nrf2、HO-1、NQO-1、Keap1蛋白及mRNA表达明显上调(P<0.05或P<0.01);与模型对照组比较,给药各组大鼠海马CA1区锥体细胞排列紧密,边界较清,细胞水肿改善;MDA含量显著降低(P<0.01)、GSH-Px活力和GSH含量明显升高(P<0.05或P<0.01),Nrf2、HO-1、NQO-1蛋白及mRNA表达显著上调,Keap1表达明显下调(P<0.05或P<0.01)。结论:大补肺汤可调控Keap1-Nrf2信号通路,减轻高原低氧大鼠脑组织氧化应激反应,对高原低氧大鼠脑损伤具有一定的保护作用。
Objective:To investigate the intervention effect of Dabufei decoction(大补肺汤) on brain injury in rats with high-altitude hypoxia by regulating the expression of Kelch-like ECH-associated protein 1(Keap1)/nuclear factor erythroid 2 related factor 2(Nrf2)signaling pathway and associated molecules. Methods:Sixty SPF grade SD rats were randomly divided into normal control group, model group, dexamethasone 0.5 mg/kg group, Dabufei decoction 8.8,17.6 and 35.1 g/kg groups. After adapting feeding for 3 days, the rats in Dabufei decoction were given corresponding drugs by intragastric administration for 14 days. The rats in dexamethasone group received the drugs by intraperitoneal injection for 3 consecutive days before entering the chamber, and the rats in other groups were given the same volume of normal saline. Except for normal control group, from the 15th day, the rats in each group were exposed to hypoxia in the hypobaric chamber for experimental animals for 3 consecutive days. The animals were sacrificed at the end of modeling, and HE staining was conducted to observe the morphology of the hippocampus. The malondialdehyde(MDA) content, glutathione peroxidase(GSH-Px) activity, and glutathione(GSH) content in the brain tissue were detected by the TBA method, colorimetric method, and microplate method, respectively. Western blotting(WB) and RT-qPCR were used to detect the protein and mRNA expressions of Keap1,Nrf2,Heme oxygenase-1(HO-1) and quinone oxidoreductase L(NQO-1) in the brain tissue of rats. Results:HE results showed that in model group, the pyramidal cells in the CA1 region of the hippocampus were arranged irregularly and swollen, with loose and light stained cytoplasm. Compared with normal control group, the activity of GSH-Px and the content of GSH were significantly decreased(P<0.01,P<0.05),while MDA content was significantly increased( P<0.01) in the brain of the model group. The protein and mRNA expressions of Nrf2,HO-1,NQO-1,and Keap1 were significantly up regulated(P<0.05 or P<0.01) in the brain of the model group. Compared with the model group, the pyramidal cells in the hippocampal CA1 region were compactly arranged with clear boundary, and cell swelling was alleviated in Dabufei decoction groups. The MDA content was decreased(P<0.01),the GSH-Px activity and GSH content were significantly increased(P<0.01,P<0.05),the protein and mRNA expressions of Nrf2,HO-1,and NQO-1 were significantly up regulated, while the Keap1 expression was significantly down regulated(P<0.01,P<0.05). Conclusion:Dabufei decoction has a protective effect on brain injury of the rats with high-altitude hypoxia by regulating the Keap1-Nrf2 signaling pathway and reducing oxidative stress in brain tissues.