蒿鳖养阴软坚方通过激活Nrf2/NQO1信号通路抑制肝纤维化发生
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摘要
目的:探讨蒿鳖养阴软坚方通过激活Nrf2/NQO1信号通路对四氯化碳复合因素所致大鼠肝纤维化的抑制作用。方法:选用健康Wistar大鼠作为研究对象,将大鼠随机分为正常组、模型组、低剂量组、中剂量组、高剂量组和秋水仙碱组。除正常组大鼠外,其余大鼠均给予自制高脂饲料饲养,每周两次皮下注射40%CC14花生油混合溶液,隔日灌胃给予30%乙醇,各给药组每日灌胃给药的同时给予正常组和模型组以等容积的蒸馏水,直至造模时间(共6周)结束后,乌拉坦麻醉大鼠并收集肝组织标本,比色法检测肝组织中羟脯氨酸(Hyp)的含量,免疫组织化学法检测各组大鼠肝组织中Nrf2和NQO1的分布情况,Western blot法检测各组大鼠肝组织中Nrf2和NQO1的表达水平。结果:与正常组相比,模型组Hyp含量明显升高(P<0.05);各给药组的Hyp含量显著低于模型组,且肝组织中Nrf2和NQO1的表达水平显著升高(P<0.05)。结论:蒿鳖养阴软坚方可以通过激活Nrf2/NQO1通路,上调Nrf2和NQO1的表达,降低肝组织中Hyp的含量,从而发挥抑制由四氯化碳复合因素所致大鼠肝纤维化的作用。
Objective: To investigate the inhibitory effect of HaobieYangyinRuanjian on activation of Nrf2/NQO1 signaling pathway on hepatic fibrosis induced by carbon tetrachloride complex. Methods: Wistar rats were randomly divided into normal group, model group,low dose group, middle dose group, high dose group and colchicine group.Except the normal group, other rats were given homemade highfat diet, subcutaneous injection of 40% CCl4 peanut oil mixed solution twice a week, and intragastric administration of 30% ethanol for 1 m L every day.The rats in the model group were treated with equal dose of distilled water until the end of the experiment for 6 weeks. Uralan rats were anesthetized and the liver tissue samples were collected. The content of hydroxyproline(Hyp) in liver tissue was detected by colorimetry. The distribution of Nrf2 and NQO1 in liver tissue was detected by chemical method. The expression of Nrf2 and NQO1 in liver tissue was detected by Western blot. Results: Compared with the normal group, the content of Hyp of the model group was significantly higher than that of the normal group(P<0.05). The expressions of Nrf2 and NQO1 in the liver tissue were significantly higher than those in the model group(P <0.05). Conclusion: The activation of Nrf2/NQO1 pathway can up-regulate the expressions of Nrf2 and NQO1 and decrease the content of Hyp in liver tissue, so as to inhibit the hepatic fibrosis induced by carbon tetrachloride complex factors.
Objective: To investigate the inhibitory effect of HaobieYangyinRuanjian on activation of Nrf2/NQO1 signaling pathway on hepatic fibrosis induced by carbon tetrachloride complex. Methods: Wistar rats were randomly divided into normal group, model group,low dose group, middle dose group, high dose group and colchicine group.Except the normal group, other rats were given homemade highfat diet, subcutaneous injection of 40% CCl4 peanut oil mixed solution twice a week, and intragastric administration of 30% ethanol for 1 m L every day.The rats in the model group were treated with equal dose of distilled water until the end of the experiment for 6 weeks. Uralan rats were anesthetized and the liver tissue samples were collected. The content of hydroxyproline(Hyp) in liver tissue was detected by colorimetry. The distribution of Nrf2 and NQO1 in liver tissue was detected by chemical method. The expression of Nrf2 and NQO1 in liver tissue was detected by Western blot. Results: Compared with the normal group, the content of Hyp of the model group was significantly higher than that of the normal group(P<0.05). The expressions of Nrf2 and NQO1 in the liver tissue were significantly higher than those in the model group(P <0.05). Conclusion: The activation of Nrf2/NQO1 pathway can up-regulate the expressions of Nrf2 and NQO1 and decrease the content of Hyp in liver tissue, so as to inhibit the hepatic fibrosis induced by carbon tetrachloride complex factors.
引文
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[15]Faig M,Bianchet M A,Winski S,et al.Structure-based development of anticancer drugs:complexes of NAD(P)H:quinoneoxidoreductase1 with chemotherapeutic quinones[J].Structure,2001,9(8):659
[2]Sun M.Kisseleva T.Reversibility of liver fibrosis[J].Clin Res Hepatol Gastroenterol,2015,39(supp11):S60
[3]Frideman S L,Bansal M B.Reversal of hepatic fibrosis-fact of fantasy[J].Hepatology,2006,43(2 suppl 1):S82
[4]Hazem S H,Shaker M E,Ashamallah S A,et al.The novel Janus kinase inhibitor ruxolitinib confers protectionagainst carbon tetrachloride-induced hepatotoxicity viamultiplemechanisms[J].Chem Biol Interact,2014,5(220):116
[5]杨凤蕊,娄建石,方步武.蒿鳖养阴软坚方抗CCl4复合因素所致大鼠肝纤维化的作用[J].中草药,2011,42(3):530
[6]邢伟,孔维涵,方步武.蒿鳖养阴软坚方对免疫性肝纤维化大鼠的治疗作用[J].中草药,2010(10):1667
[7]率红莉,方步武,邢伟,等.蒿鳖养阴软坚方对四氯化碳复合因素所致肝纤维化的预防作用[J].中国实验方剂学杂志,2013,19(16):197
[8]寇咏梅,刘洁莹,方步武.蒿鳖养阴软坚方抗小鼠血吸虫病肝纤维化及其机制的研究[J].现代药物与临床,2013(3):288
[9]冯硕,万永臻,张玉萍,等.蒿鳖养阴软坚方给药大鼠体内乙醛含量的测定[J].现代中药研究与实践,2011,25(4):44
[10]Han X Y,Hu J N,Wang Z,et al.5-HMF attenuates liver fibrosis in CCl4-plus-alcohol-induced mice by suppression of oxidative stress[J].J Nutr Sci Vitaminol,2017,63(1):35
[11]Xu J Y,Su Y Y,Cheng J S,et al.Hepatoprotective effects of fullerenol on carbon tetrachloride-induced acute hepatotoxicityand nephrotoxicity in rats[J].Carbon,2010,48(5):1388
[12]Shi F,Li X,Pan H,et al.NQO1 and CYP450 reductase decrease the systemic exposure of rifampicin-quinone and mediate its redox cycle in rats[J].J Pharm Biomed Anal,2017,5(132):17
[13]Elaify M S,Adly A A,Attia A A,et al.Effect of antioxidant therapy on hepatic fibrosis and liver iron concentrations inβ-thalassemia major patients[J].Hemoglobin,2013,37(3):257
[14]Wu K C,Cui J Y,Klaassen C D.Effect of graded Nrf2 activation on phase-I and-II drug metabolizing enzymes and transportersin mouse liver[J].PLo S One,2012,7(7):e39006
[15]Faig M,Bianchet M A,Winski S,et al.Structure-based development of anticancer drugs:complexes of NAD(P)H:quinoneoxidoreductase1 with chemotherapeutic quinones[J].Structure,2001,9(8):659