摘要
目的:探讨缺氧不同时间对肝脏氧化应激水平的影响及肝肾功能发生的变化,研究黄芪甲苷基于Nrf2/HO-1通路对模拟急进高原大鼠肝脏的保护性作用。方法:建立模拟高原急性缺氧动物模型,56只SD大鼠随机分为对照组(Control),缺氧组(H),黄芪甲苷组(AS-IV+H),H组和AS-IV+H组进一步分为12 h,24 h和48 h亚组,每组8只,共7组。AS-IV+H组给予黄芪甲苷灌胃(80 mg/kg),1次/d,连续7d。将实验组大鼠放入低压氧舱(模拟海拔5000 m),到达预定时间后,分别检测大鼠血清肝肾功,肝组织中超氧化物歧化酶(SOD),丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-PX)的含量;蛋白印记法和荧光定量PCR分别检测肝脏组织中Nrf2、HO-1蛋白表达及其基因表达水平;肝组织的病理学表现通过HE染色观察。结果:与常氧环境的对照组比较,不同缺氧时间段的H组和AS-IV+H组肝肾功指标均高于对照组,且随缺氧时间的增加,呈现增高的趋势(P<0.05或P<0.01)。组织匀浆中抗氧化酶除MDA高于对照组外,其余抗氧化酶超氧SOD、GSH-PX水平均低于对照组(P<0.05或P<0.01)。肝组织中Nrf2、HO-1蛋白及mRNA表达随缺氧时间的延长而升高,在缺氧24h达到高峰,缺氧48h表达较缺氧24h有所下降,但在各时间段均高于对照组(P<0.05或P<0.01)。与同时间缺氧组比较,黄芪甲苷组肝肾功,SOD,GSH-PX均低于同时间缺氧,但MDA高于同时间缺氧组(P<0.05或P<0.01)。肝组织中Nrf2、HO-1蛋白及mRNA表达均高于同时间缺氧组(P<0.05或P<0.01)。HE染色显示缺氧组肝细胞水肿,黄芪甲苷组水肿减轻。结论:急进高原缺氧环境导致肝肾功能出现异常,肝脏组织发生相关的病理学改变,黄芪甲苷干预能提高高原低氧环境下肝脏对抗氧化应激损伤的能力,保护低氧环境下的肝肾功能,提高机体在高原低氧环境下的适应性能力。
Objective:To investigate the effects of hypoxia at different time on the level of liver oxidative stress and the changes in liver and kidney function,and to study the protective effect of astragaloside IV on the liver of rats simulated acute plateau exposure based on the Nrf2/HO-1 pathway.Methods:An animal model of simulated acute plateau hypoxia was established.56 SD rats were randomly divided into a control group(Control),a hypoxia group(H),and an astragaloside IV group(AS-IV+H).The H group and the AS-IV+H group were further divided into 12 h,24 h,and 48 h subgroups,with 8 rats in each group,for a total of 7 groups.The AS-IV+H group was given astragaloside IV by gavage(80 mg/kg),once a day,for 7 consecutive days.The rats in the experimental group were placed in a hypobaric oxygen chamber(simulating an altitude of 5000 m).After the predetermined time,the liver and kidney function of the rats in serum,the content of superoxide dismutase(SOD),malondialdehyde(MDA),and glutathione peroxidase(GSH-PX)in liver tissue were detected respectively;the protein expression and gene expression level of Nrf2 and HO-1 in liver tissue were detected by Western blotting and fluorescence quantitative PCR respectively;the pathological manifestations of liver tissue were observed by HE staining.Results:Compared with the control group in normoxic environment,the liver and kidney function indexes of the H group and AS-IV+H group at different hypoxic time periods were higher than those of the control group,and showed an increasing trend with the increase of hypoxia time(P<0.05,P<0.01).In the tissue homogenate,except for MDA,the levels of other antioxidant enzymes,superoxide SOD and GSH-PX,were lower than those of the control group(P<0.05 or P<0.01).The expression of Nrf2 and HO-1 protein and mRNA in liver tissue increased with the extension of hypoxia time,reaching the peak at 24h hypoxia,and decreased at 48h hypoxia compared with 24h hypoxia,but was higher than that in the control group at all time periods(P<0.05,P<0.01).Compared with the hypoxia group at the same time,the liver and kidney function,SOD,and GSH-PX in the astragaloside IV group were lower than those in the hypoxia group at the same time,but MDA was higher than those in the hypoxia group at the same time(P<0.05,P<0.01).The expression of Nrf2 and HO-1 protein and mRNA in liver tissue were higher than those in the hypoxia group at the same time(P<0.05,P<0.01).HE staining showed that the hepatocytes in the hypoxia group were edematous,and the edema in the astragaloside IV group was alleviated.Conclusion:Rapid exposure to high altitude hypoxia leads to abnormal liver and kidney function and pathological changes in liver tissue.Astragaloside IV intervention can improve the liver’s ability to resist oxidative stress damage in high altitude hypoxia,protect liver and kidney function in hypoxia,and improve the body’s adaptability in high altitude hypoxia.
作者
王娅鑫
罗晓红
董俐
申栋帅
杜军
许瑞元
肖攀
WANG Yaxin;LUO Xiaohong;DONG Li;SHEN Dongshuai;DU Jun;XU Ruiyuan;XIAO Pan(No.940 Hospital of Joint Service Support Force of Chinese People’s Liberation Army,Gansu Lanzhou 730050,China;Gansu University of Traditional Chinese Medicine,Gansu Lanzhou 730030,China;Basic Medical Laboratory,No.940 Hospital of Joint Service Support For ce of Chinese people′s Liberation Army,Gansu Lanzhou 730050,China)
出处
《中医药临床杂志》
2024年第9期1745-1751,共7页
Clinical Journal of Traditional Chinese Medicine
基金
甘肃省自然科学基金(22JR5RA023)
军队实验动物专项课题(SYDW〔2020〕27号)
面上项目(2021yxky040)。
关键词
高原低氧
氧化应激
NRF2
HO-1
肝肾功能
黄芪甲苷
High altitude hypoxia
Oxidative stress
Nrf2
HO-1
Liver and kidney function
Astragaloside IV
