加味天王补心丹对模拟高原暴露小鼠的抗缺氧作用研究  

Study on the Anti-hypoxic Effect of Flavored Tianwang Buxin Dan on Mice Exposed to Simulated Plateau Environment

作　　者：黄婷[1,2] 李硕 高安然 谭小翠 李劲草[2] 薛瑞 张杨 李迎 王萧 张有志 HUANG Ting;LI Shuo;GAO Anran;TAN Xiaocui;LI Jincao;XUE Rui;ZHANG Yang;LI Ying;WANG Xiao;ZHANG Youzhi(Laboratory Animal Center,Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;Institute of Pharmacology and Toxicology,Academy of Military Medical Sciences,State Key Laboratory of Toxicology and Medical Countermeasures,Beijing 100850,China)

机构地区：[1]广州中医药大学动物实验中心,广东广州510405 [2]军事科学院军事医学研究院毒物药物研究所,抗毒药物与毒理学国家重点实验室,北京100850

出　　处：《中药新药与临床药理》2022年第9期1156-1162,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：广东省普通高校特色创新项目(2019KTSCX030);广东省基础与应用基础研究基金自然科学基金项目(2021A1515012524);广州市科技计划项目(202002030433)。

摘　　要：目的 探讨加味天王补心丹(JWBXD)对模拟高原暴露小鼠抗缺氧能力的影响及其作用机制。方法将雄性C57小鼠随机分为正常组、模型组、JWBXD低剂量组(0.51 g·kg^(-1))、JWBXD高剂量组(1.02 g·kg^(-1))、红景天组(0.40 g·kg^(-1)),灌胃给药,每日1次,连续21 d。给药结束后,除模型组外,各组均随机选取8只小鼠分别进行常压耐缺氧实验;给药14 d后,各组均随机选取8只小鼠分别进行低压耐缺氧实验。常压耐缺氧实验中,观察密闭缺氧实验和亚硝酸钠(230 mg·kg^(-1))中毒实验中小鼠的存活时间,并计算小鼠存活时间延长率。低压耐缺氧实验中,小鼠放入模拟高原5 500 m的低压低氧动物实验舱内,连续造模7 d;采用苏木精-伊红(HE)染色法观察小鼠脑海马体及肺组织病理变化;采用黄嘌呤氧化酶法检测脑组织中超氧化物歧化酶(SOD)活性;采用硫代巴比妥酸比色法检测脑组织中丙二酫(MDA)含量;Western Blot法检测心肌组织HIF-1α、p-AMPK/AMPK、PGC-1α及Nrf2蛋白表达。结果 在常压耐缺氧实验中,与正常组相比,各给药组小鼠的密闭缺氧存活时间及亚硝酸钠中毒后存活时间均明显延长(P<0.05,P<0.01),存活时间延长率明显提高。在低压耐缺氧实验中,与正常组比较,模型组小鼠的脑海马体、肺组织出现明显的病理损伤;脑组织中的SOD活性明显降低(P<0.01),MDA含量明显升高(P<0.01);心肌组织中的HIF-1α蛋白表达明显上调(P<0.05),p-AMPK/AMPK、PGC-1α及Nrf2蛋白表达明显下调(P<0.05,P<0.01)。与模型组比较,各给药组小鼠的脑海马体、肺组织的病理损伤明显减轻;脑组织中的SOD活性均明显升高(P<0.01),MDA含量均明显降低(P<0.01);心肌组织中的HIF-1α、p-AMPK/AMPK、PGC-1α及Nrf2蛋白表达均明显上调(P<0.05,P<0.01)。结论 JWBXD能够提高模拟高原暴露小鼠的抗缺氧能力,改善高原环境下小鼠的氧化应激损伤,其机制可能与提高机体抗氧化能力,激活HIF-1α/AObjective To investigate the effect of flavored Tianwang Buxin Dan(JWBXD) on the anti-hypoxic ability of mice exposed to simulated plateau environment and its mechanism of action. Methods Male C57 mice were randomly divided into normal group,model group,JWBXD low-dose group(0.51 g·kg^(-1)),JWBXD high-dose group(1.02 g·kg^(-1)) and Herba Rhodiolae group(positive control,0.40 g·kg^(-1)) and administered by gavage once daily for consecutive 21 days. At the end of administration,8 mice in each group were randomly selected for normbaric hypoxia tolerance experiment except the model group;after 14 days of administration,8 mice in each group were randomly selected for the low-pressure hypoxia test. In the normobaric hypoxia experiment,the survival time of mice in the closed hypoxia experiment and the sodium nitrite(230 mg·kg^(-1)) poisoning test were observed,and the extension rate of survival time was calculated. In the low-pressure hypoxia tolerance test,mice were placed in a low-pressure hypoxic chamber at 5 500-meter on a simulated plateau environment for consecutive 7 days;the pathological changes in the hippocampus and lung tissues of mice were observed by HE staining method;the activity of SOD in brain tissue was detected by xanthine oxidase method;the content of MDA in brain tissue was detected by thiobarbituric acid colorimetry;and the protein expressions of HIF-1α,p-AMPK/AMPK,PGC-1α and Nrf2 in myocardial tissues were detected by Western Blot. Results In the normobaric hypoxia tolerance experiment,compared with the normal group, the survival time of closed hypoxia and the survival time after sodium nitrite poisoning were significantly prolonged(P<0.05,P<0.01),and the survival time extension rate was significantly increased in each administration group. In the low-pressure hypoxia tolerance experiment,the cerebral hippocampus and lung tissues of the model group showed obvious pathological damage compared with normal group;the SOD activity in brain tissues was significantly decreased(P<0.01) and the MDA co

关 键 词：加味天王补心丹 模拟高原环境 抗缺氧 氧化应激 HIF-1α/AMPK信号通路 小鼠 

分 类 号：R285.5[医药卫生—中药学]