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  • Title: [Herbal cake-separated moxibustion relieves aspirin induced gastric mucosal injury by activating Nrf2/ARE/HO-1 signaling in rats].
    Author: Yi Z, Bi DY, Xu Y, Wang JY.
    Journal: Zhen Ci Yan Jiu; 2023 Aug 25; 48(8):773-81. PubMed ID: 37614135.
    Abstract:
    OBJECTIVE: To investigate whether herbal cake-separated moxibustion can activate nuclear factor erythroid 2-related factor 2 （Nrf2） / antioxidant responsive element （ARE） /hemeoxygenase-1 （HO-1） signaling pathway to repair aspirin induced gastric mucosal injury （GMI） in rats. METHODS: SD rats （half male and half female） were randomly divided into blank control, model, moxibustion, inhibitor of HO-1 （inhibitor）, model+inhibitor, moxibustion + inhibitor groups, with 20 rats in each group. The GMI model was established by gavage of aspirin 150 mg/kg（1 mL/100 g）. Herbal cake-separated moxibustion was alternatively applied to bilateral "Zusanli" （ST36） and "Zhongwan" （CV12） and bilateral "Pishu" （BL20） and "Weishu" （BL21） for 30 min, once daily for 8 days. The rats in the three inhibitor groups received intraperitoneal injection of HO-1 inhibitor zinc protoporphyrin （5 mg/kg）. The rats' behavior score, emotional response score, skin hair score, diet score and stool state score were given. The GMI index was calculated according to Guth's methods. Histopathological changes of gastric mucosa were observed by H.E. staining. The activity of superoxide dismutase （SOD） and the content of malondialdehyde （MDA） in the gastric mucosal tissue and serum were detected by ELISA. The levels of Nrf2/ARE/HO-1 signaling pathway-related factors Keap1, Nrf2, HO-1, CAT, GST, and NQO1 in the gastric mucosal tissue were detected by quantitative real-time PCR and Western blot, separately. RESULTS: Compared with the blank control group, the behavior score, emotional response score, skin hair score, diet score and stool state score, GMI index, MDA contents of gastric mucosal tissue and serum, expression le-vels of Keap1 mRNA and protein were significantly increased （P<0.01）, while the activity of gastric mucosal and serum SOD, the expression levels of Nrf2, HO-1, CAT, GST and NQO1 mRNAs and proteins were considerably decreased （P<0.01） in the model group. Compared with the model group, moxibustion obviously reversed the increase of emotional response score, skin hair score, stool state score, GMI index, MDA levels of gastric mucosal tissue and serum, and expression levels of Keap1 mRNA and protein （P<0.01, P<0.05）, and the decrease of activity of SOD of gastric mucosal and serum, and the expression levels of Nrf2, HO-1, CAT, GST and NQO1 mRNAs and proteins （P<0.01, P<0.05）. After administration of antagonist of HO-1, the effects of moxibustion were eliminated or weakened pronouncedly in reducing skin hair score, GMI index, contents of gastric mucosal and serum MDA, and expression of Keap1 mRNA and protein, and in up-regulating gastric mucosal and serum SOD, and expression of HO-1, CAT, GST and NQO1 mRNAs and proteins （P<0.01, P<0.05）. CONCLUSION: Herbal-cake separated moxibustion can improve the GMI in rats, which may be associated with its effects in reducing oxidative stress and activating Nrf2/ARE/HO-1 signaling pathway. 目的：探讨隔药饼灸是否通过激活核因子E2相关因子2（Nrf2）/抗氧化反应元件（ARE）/血红素加氧酶-1（HO-1）信号通路修复阿司匹林诱导的胃黏膜损伤。方法：SD大鼠随机分为空白对照组、模型组、隔药饼灸穴位组、空白+抑制剂组、模型+抑制剂组、隔药饼灸+抑制剂组，每组20只。阿司匹林（150 mg/kg）灌胃诱导复制胃黏膜损伤模型。隔药饼灸穴位组和隔药饼灸+抑制剂组施隔药饼灸“足三里”“中脘”“脾俞”“胃俞”，每次30 min，连续治疗8 d。3个抑制剂组的大鼠腹腔内注射HO-1抑制剂锌原卟啉 5 mg/kg。观察各组大鼠外观行为变化并进行评分；观察胃黏膜损伤程度并对胃黏膜损伤指数进行评定；HE染色观察胃黏膜组织形态学变化；ELISA法检测胃黏膜组织、血清中超氧化物歧化酶（SOD）活性和丙二醛（MDA）含量。荧光定量PCR法和Western blot法检测胃黏膜组织中Nrf2/ARE/HO-1信号通路相关因子Keap1、Nrf2、HO-1、过氧化氢酶（CAT）、谷胱甘肽S-转移酶（GST）、NAD（P）H醌脱氢酶1（NQO1） mRNA和蛋白表达水平。结果：与空白对照组比较，模型组大鼠的行为状态、情绪反应、皮肤毛发、饮食状态和大便状态评分升高（P<0.01），胃黏膜炎性反应明显，胃黏膜损伤指数升高（P<0.01），胃黏膜组织和血清中SOD活性降低（P<0.01），MDA含量升高（P<0.01），胃黏膜组织中Keap1 mRNA和蛋白表达上调（P<0.01），Nrf2、HO-1、CAT、GST、NQO1 mRNA和蛋白表达下调（P<0.01）。与模型组比较，隔药饼灸穴位组大鼠的情绪反应、皮肤毛发和大便状态评分降低（P<0.01，P<0.05），胃黏膜损伤指数降低（P<0.01），胃黏膜炎性反应减轻，胃黏膜组织和血清中SOD活性升高（P<0.01），MDA含量降低（P<0.01），Keap1 mRNA和蛋白表达下调（P<0.01），Nrf2、HO-1、CAT、GST、NQO1 mRNA和蛋白表达上调（P<0.05，P<0.01）。隔药饼灸+抑制剂组大鼠使用HO-1抑制剂后，与隔药饼灸穴位组比较，皮肤毛发评分升高（P<0.01），胃黏膜损伤指数升高（P<0.05），胃黏膜炎性反应加重，胃黏膜组织和血清中SOD活性降低（P<0.01），MDA含量升高（P<0.01），Keap1 mRNA和蛋白表达上调（P<0.05），HO-1、CAT、GST、NQO1 mRNA和蛋白表达下调 （P<0.05，P<0.01），Nrf2蛋白表达下调（P<0.05）。结论：隔药饼灸可通过激活Nrf2/ARE/HO-1信号通路，降低阿司匹林诱导的胃黏膜细胞的氧化应激，从而修复胃黏膜损伤。.
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