海藻玉壶汤中海藻不同品种与甘草加减应用对甲状腺肿大大鼠氧化应激及肝脏Nrf2/HO-1通路的影响
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摘要
目的探讨海藻玉壶汤中海藻不同品种与甘草加减应用对甲状腺肿大大鼠氧化应激及肝脏核因子E2相关因子2/血红素加氧酶-1(Nrf2/HO-1)通路的影响。方法随机分为空白组、模型组、阳性药优甲乐组(优甲乐组)、含羊栖菜的海藻玉壶汤全方组(羊栖菜全方组)、含海蒿子的海藻玉壶汤全方组(海蒿子全方组)、含羊栖菜的海藻玉壶汤去甘草组(羊栖菜去甘草组)、含海蒿子的海藻玉壶汤去甘草组(海蒿子去甘草组)、海藻玉壶汤去海藻组(全方去海藻组)、海藻玉壶汤去海藻甘草组(全方去海藻甘草组)。除空白组外其余各组均灌服丙硫氧嘧啶(PTU)复制甲状腺肿大模型,以优甲乐作为阳性对照药。检测各组大鼠血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、丙二醛(MDA)、过氧化氢(H_2O_2)、黄嘌呤氧化酶(XOD)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)的含量,测定肝组织Nrf2 mRNA及HO-1 mRNA的表达。结果与空白组比较,模型组血清MDA升高、Nrf2 mRNA表达降低(P<0.01),海蒿子去甘草组血清ALT、肝组织HO-1 mRNA表达升高(P<0.05,P<0.01),海蒿子全方组血清MDA、肝组织HO-1 mRNA表达升高(P<0.05,P<0.01),羊栖菜去甘草组及羊栖菜全方组血清ALT、AST、MDA、H_2O_2升高,GSH-Px、Nrf2 mRNA表达降低(P<0.05,P<0.01);与海蒿子去甘草组比较,羊栖菜全方组血清AST升高、肝组织HO-1 mRNA表达降低(P<0.05,P<0.01),羊栖菜去甘草组血清H_2O_2升高、HO-1 mRNA表达降低(P<0.05,P<0.01);与海蒿子全方组比较,羊栖菜去甘草组血清ALT、AST、H_2O_2升高,HO-1 mRNA表达降低(P<0.05,P<0.01),羊栖菜全方组血清AST升高、HO-1 mRNA表达降低(P<0.01)。结论不同品种海藻(海蒿子/羊栖菜)与甘草在海藻玉壶汤中加减应用,海蒿子全方组及海蒿子去甘草组在一定程度上可以保护肝脏组织免受过氧化物的损害,其机制可能与激活Nrf2/HO-1通路提高肝细胞抗氧化能力有关;而羊栖菜全方组及羊栖菜去甘草组肝功能受到一定影响,可能与氧化抗氧化系统平衡紊乱,Nrf2/HO-1通路激活障碍有关。
Objective To investigate the influence of different species, Haihaozi(Alga Sargassi Pallidi, Sargassum pallidum) and Yangqicai(Alga Sargassi Fusiformis, Sargassum fusiforme),of Haizao(Algae, Sargassum) and modified administration of Gancao(Liquorice Root, Radix Glycyrrhizae) in Haizao Yuhu Tang(Algae Okho Decoction, HZYH Decoction) containing antagonism combination on oxidative stress and liver Nrf2/HO-1 pathway in rats with goiter. Methods All rats(n=144) were randomly divided into blank group, model group, positive drug euthyrox group(euthyrox group), HZYH Decoction with Yangqicai group, HZYH Decoction with Haihaozi group, HZYH Decoction with Yangqicai without Gancao group, HZYH Decoction with Haihaozi without Gancao group, HZYH Decoction without Haizao group, and HZYH Decoction without Haizao and Gancao group. Except of blank group, all other groups were orally given PTU for copying goiter model. Euthyrox was taken as positive control drug. The content of serum ALT, AST, MDA, H_2O_2, XOD, SOD and GSH-Px were detected in all groups. The expressions of Nrf2 mRNA HO-1 mRNA in liver tissue were detected. Results The content of MDA increased and expression of Nrf2 mRNA decreased in model group compared with blank group(P<0.01). The content of ALT and expression of HO-1 mRNA increased in HZYH Decoction with Haihaozi without Gancao group(P<0.05, P<0.01) and expression of HO-1 mRNA increased in HZYH Decoction with Haihaozi group(P<0.01). The content of ALT, AST, MDA and H_2O_2 increased, and GSH-Px content and expression of Nrf2 mRNA decreased in HZYH Decoction with Yangqicai without Gancao group and HZYH Decoction with Yangqicai group(P<0.05, P<0.01). The content of AST increased and expression of HO-1 mRNA had no significant changes in HZYH Decoction with Yangqicai group compared with HZYH Decoction with Yangqicai without Gancao group(P<0.05, P<0.01). The content of H_2O_2 increased and expression of HO-1 mRNA had no significant changes in HZYH Decoction with Yangqicai without Gancao group(P<0.05, P<0.01). The content of ALT, AST and H_2O_2 increased and expression of HO-1 mRNA had no significant changes in HZYH Decoction with Yangqicai without Gancao group compared with HZYH Decoction with Haihaozi group(P<0.05, P<0.01). The content of AST increased and expression of HO-1 mRNA had no significant changes in HZYH Decoction with Yangqicai group(P<0.01). Conclusion The modified administration of different species of Haizao(Haihaozi/Yangqicai) in HZYJ Decoction, and HZYH Decoction with Haihaozi group and HZYH Decoction with Haihaozi without Gancao group can protect liver tissue to some extent from peroxide injury. The mechanism may be related to activation of Nrf2/HO-1 pathway and improvement of antioxidant ability of liver cells. The liver function is affected in HZYH Decoction with Yangqicai group and HZYH Decoction with Yangqicai without Gancao group, which may be related to disturbance of oxidant-antioxidant system and failure of activating Nrf2/HO-1 pathway.
Objective To investigate the influence of different species, Haihaozi(Alga Sargassi Pallidi, Sargassum pallidum) and Yangqicai(Alga Sargassi Fusiformis, Sargassum fusiforme),of Haizao(Algae, Sargassum) and modified administration of Gancao(Liquorice Root, Radix Glycyrrhizae) in Haizao Yuhu Tang(Algae Okho Decoction, HZYH Decoction) containing antagonism combination on oxidative stress and liver Nrf2/HO-1 pathway in rats with goiter. Methods All rats(n=144) were randomly divided into blank group, model group, positive drug euthyrox group(euthyrox group), HZYH Decoction with Yangqicai group, HZYH Decoction with Haihaozi group, HZYH Decoction with Yangqicai without Gancao group, HZYH Decoction with Haihaozi without Gancao group, HZYH Decoction without Haizao group, and HZYH Decoction without Haizao and Gancao group. Except of blank group, all other groups were orally given PTU for copying goiter model. Euthyrox was taken as positive control drug. The content of serum ALT, AST, MDA, H_2O_2, XOD, SOD and GSH-Px were detected in all groups. The expressions of Nrf2 mRNA HO-1 mRNA in liver tissue were detected. Results The content of MDA increased and expression of Nrf2 mRNA decreased in model group compared with blank group(P<0.01). The content of ALT and expression of HO-1 mRNA increased in HZYH Decoction with Haihaozi without Gancao group(P<0.05, P<0.01) and expression of HO-1 mRNA increased in HZYH Decoction with Haihaozi group(P<0.01). The content of ALT, AST, MDA and H_2O_2 increased, and GSH-Px content and expression of Nrf2 mRNA decreased in HZYH Decoction with Yangqicai without Gancao group and HZYH Decoction with Yangqicai group(P<0.05, P<0.01). The content of AST increased and expression of HO-1 mRNA had no significant changes in HZYH Decoction with Yangqicai group compared with HZYH Decoction with Yangqicai without Gancao group(P<0.05, P<0.01). The content of H_2O_2 increased and expression of HO-1 mRNA had no significant changes in HZYH Decoction with Yangqicai without Gancao group(P<0.05, P<0.01). The content of ALT, AST and H_2O_2 increased and expression of HO-1 mRNA had no significant changes in HZYH Decoction with Yangqicai without Gancao group compared with HZYH Decoction with Haihaozi group(P<0.05, P<0.01). The content of AST increased and expression of HO-1 mRNA had no significant changes in HZYH Decoction with Yangqicai group(P<0.01). Conclusion The modified administration of different species of Haizao(Haihaozi/Yangqicai) in HZYJ Decoction, and HZYH Decoction with Haihaozi group and HZYH Decoction with Haihaozi without Gancao group can protect liver tissue to some extent from peroxide injury. The mechanism may be related to activation of Nrf2/HO-1 pathway and improvement of antioxidant ability of liver cells. The liver function is affected in HZYH Decoction with Yangqicai group and HZYH Decoction with Yangqicai without Gancao group, which may be related to disturbance of oxidant-antioxidant system and failure of activating Nrf2/HO-1 pathway.
引文
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[10] 王昕,姚凝,刘建鸿,等. 甘草与海藻配伍对小鼠肝脏的毒理作用及氧化-抗氧化平衡研究[J]. 时珍国医国药,2012,23(4):879-880.Wang X, Yao N, Liu JH, et al. Toxicological effects and oxidation-antioxidant balance of combined Herba Sargassum and Glycyrrhiza Uralensis in mouse liver[J]. Lishizhen Medicine and Materia Medica Research, 2012, 23(4): 879-880.
[11] 孟娴.中药海藻与甘草配伍致毒增毒作用机理研究[D]. 南京:南京中医药大学,2018.Meng X. Study on the mechanism of toxicity of the Sargassum-Glycyrrhiza Uralensis extract[D]. Nanjing: Nanjing University of Chinese Medicine, 2018.
[12] 李怡文.含海藻与甘草反药组合的海藻玉壶汤应用于甲状腺肿大大鼠模型配伍宜忌条件研究[D]. 北京:北京中医药大学,2013:75-76.Li YW. Study on compatibility conditions of Haizao Yuhu Tang containing incompatible medicinal combination of Haizao and Gancao in rat model of thyroid enlargement[D]. Beijing: Beijing University of Traditional Chinese Medicine, 2013: 75-76.
[13] 修琳琳.含反药组合的海藻玉壶汤中海藻不同品种与甘草加减对甲状腺肿大大鼠生物效应影响及机制探讨[D]. 北京:北京中医药大学,2017:95-99.Xiu LL. Biological effect and mechanism of different species of Sargassum and Glycyrrhiza uralensis in Haizao Yuhu Tang containing incompatible medicinal combination in goiter rats[D]. Beijing: Beijing University of Traditional Chinese Medicine, 2017: 95-99.
[14] 杨振凯,杨阳,张亚男,等. 草苁蓉正丁醇及水萃取物对小鼠急性肝损伤的保护作用[J]. 延边大学医学学报,2011,34(2):105-107.Yang ZK, Yang Y, Zhang YN, et al. Protective effects of butanol soluble and aqueous fractions of Boschniakia rossica on acute liver injury in mice[J]. Journal of Medical Science Yanbian University, 2011, 34(2): 105-107.
[15] 佘颜,易健,邵乐,等.补阳还五汤精简方对局灶性脑缺血大鼠脑谷胱甘肽抗氧化系统的影响[J]. 中国中医药信息杂志,2015,22(1):58-61.She Y, Yi J, Shao L, et al. Effecst of thin recipe of Buyang Huanwu Decoction on expressions of GSH, GSH-Px and γ-GCS in the brain of focal cerebral ischemia rats[J]. Chinese Journal of Information on TCM, 2015, 22(1): 58-61.
[16] 张愔,刘红春,刘新郑,等. 三种肝病患者血清黄嘌呤氧化酶的变化[J]. 河南医科大学学报,1998,33(1):81-82.Zhang Y, Liu HC, Liu XZ, et al. Changes of serum xanthine oxidase in patients with three kinds of liver diseases[J]. Journal of Henan Medical University, 1998, 33(1): 81-82.
[17] Vargas MR, Johnson JA. The Nrf2-ARE cytoprotective pathway in astrocytes[J]. Expert Reviews in Molecular Medicine, 2009, 11: e17.
[18] Nguyen T, Yang CS, Pickett CB. The pathways and molecular mechanisms regulating Nrf2 activation in response to chemical stress[J]. Free Radical Biology and Medicine, 2004, 37(4): 433-441.
[19] McCubrey JA, LaHair MM, Franklin RA. Reactive oxygen species-induced activation of the MAP kinase signaling pathways[J]. Antioxid & Redox Signaling, 2006, 8(9-10): 1775-1789.
[20] Tagawa A, Kaneko T, Shinohara T, et al. Heme oxygenase-1 inhibits cigarette smoke-induced in the tracheal mucosal permeahility in guinea pigs in vivo[J]. Inflammation Research, 2005, 54(5): 229-234.
[2] Li C, Li X, You L, et al. Fractionation, preliminary structural characterization and bioactivities of polysaccharides from Sargassum pallidum[J]. Carbohydrate Polymers, 2017, 155: 261-270.
[3] Krivoshapko O A, Popov A M. Medical and prophylactic properties lipids and antioxidants derived from sea hydrobionts[J]. Voprosy Pitaniia, 2011, 80(2): 4-8.
[4] Chen P, He D, Zhang Y, et al. Sargassum fusiforme polysaccharides activate antioxidant defense by promoting Nrf2-dependent cytoprotection and ameliorate stress insult during aging[J]. Food&Function, 2016, 7(11): 4576-4588.
[5] 陈实功.外科正宗[M]. 北京:中国中医药出版社,2011:112.Chen SG. Waike Zhengzong[M]. Beijing: China Press of Traditional Chinese Medicine, 2011: 112.
[6] 张民庆.现代临床方剂学[M]. 北京:人民卫生出版社,2004:817.Zhang MQ. Modern Clinical TCM Formulas[M]. Beijing: People’s Medical Publishing House, 2004: 817.
[7] 乔丽杰,王延让,张明.Nrf2/HO-1通路在氧化损伤保护机制中研究进展[J]. 中国职业医学,2013,40(1):82-84.Qiao LJ, Wang YR, Zhang M. Research progress of Nrf2/HO-1 pathway in mechanism of oxidative damage and protection[J]. Chin Occul Med, 2013, 40(1): 82-84.
[8] 丁选胜,李欧,阚毓铭.海藻、甘草及其不同比例配伍后的水提取物的肝毒性研究[J]. 南京中医药大学学报,2003,19(1):28-31.Ding XS, Li O, Kan YM. Hepatotoxicity of Herba Sargassum and Radix Glycyrrhiza Uralensis and water extract of their combination[J]. Nanjing Univ Tradit Chin Med, 2003, 19(1): 28-31.
[9] 丁选胜,李欧.海藻甘草及其相配伍后的水提取物的肝毒性研究—对离体大鼠肝灌流液中黄嘌呤氧化酶的影响[J]. 江苏中医药,2002,23(10):51-54.Ding XS, Li O. Hepatotoxicity of Herba Sargassum and Glycyrrhiza Uralensis and their combination water extracts-effect on xanthine oxidase in isolated rat hepatic perfusate[J]. Jiangsu Journal of Traditional Chinese Medicine, 2002, 23(10): 51-54.
[10] 王昕,姚凝,刘建鸿,等. 甘草与海藻配伍对小鼠肝脏的毒理作用及氧化-抗氧化平衡研究[J]. 时珍国医国药,2012,23(4):879-880.Wang X, Yao N, Liu JH, et al. Toxicological effects and oxidation-antioxidant balance of combined Herba Sargassum and Glycyrrhiza Uralensis in mouse liver[J]. Lishizhen Medicine and Materia Medica Research, 2012, 23(4): 879-880.
[11] 孟娴.中药海藻与甘草配伍致毒增毒作用机理研究[D]. 南京:南京中医药大学,2018.Meng X. Study on the mechanism of toxicity of the Sargassum-Glycyrrhiza Uralensis extract[D]. Nanjing: Nanjing University of Chinese Medicine, 2018.
[12] 李怡文.含海藻与甘草反药组合的海藻玉壶汤应用于甲状腺肿大大鼠模型配伍宜忌条件研究[D]. 北京:北京中医药大学,2013:75-76.Li YW. Study on compatibility conditions of Haizao Yuhu Tang containing incompatible medicinal combination of Haizao and Gancao in rat model of thyroid enlargement[D]. Beijing: Beijing University of Traditional Chinese Medicine, 2013: 75-76.
[13] 修琳琳.含反药组合的海藻玉壶汤中海藻不同品种与甘草加减对甲状腺肿大大鼠生物效应影响及机制探讨[D]. 北京:北京中医药大学,2017:95-99.Xiu LL. Biological effect and mechanism of different species of Sargassum and Glycyrrhiza uralensis in Haizao Yuhu Tang containing incompatible medicinal combination in goiter rats[D]. Beijing: Beijing University of Traditional Chinese Medicine, 2017: 95-99.
[14] 杨振凯,杨阳,张亚男,等. 草苁蓉正丁醇及水萃取物对小鼠急性肝损伤的保护作用[J]. 延边大学医学学报,2011,34(2):105-107.Yang ZK, Yang Y, Zhang YN, et al. Protective effects of butanol soluble and aqueous fractions of Boschniakia rossica on acute liver injury in mice[J]. Journal of Medical Science Yanbian University, 2011, 34(2): 105-107.
[15] 佘颜,易健,邵乐,等.补阳还五汤精简方对局灶性脑缺血大鼠脑谷胱甘肽抗氧化系统的影响[J]. 中国中医药信息杂志,2015,22(1):58-61.She Y, Yi J, Shao L, et al. Effecst of thin recipe of Buyang Huanwu Decoction on expressions of GSH, GSH-Px and γ-GCS in the brain of focal cerebral ischemia rats[J]. Chinese Journal of Information on TCM, 2015, 22(1): 58-61.
[16] 张愔,刘红春,刘新郑,等. 三种肝病患者血清黄嘌呤氧化酶的变化[J]. 河南医科大学学报,1998,33(1):81-82.Zhang Y, Liu HC, Liu XZ, et al. Changes of serum xanthine oxidase in patients with three kinds of liver diseases[J]. Journal of Henan Medical University, 1998, 33(1): 81-82.
[17] Vargas MR, Johnson JA. The Nrf2-ARE cytoprotective pathway in astrocytes[J]. Expert Reviews in Molecular Medicine, 2009, 11: e17.
[18] Nguyen T, Yang CS, Pickett CB. The pathways and molecular mechanisms regulating Nrf2 activation in response to chemical stress[J]. Free Radical Biology and Medicine, 2004, 37(4): 433-441.
[19] McCubrey JA, LaHair MM, Franklin RA. Reactive oxygen species-induced activation of the MAP kinase signaling pathways[J]. Antioxid & Redox Signaling, 2006, 8(9-10): 1775-1789.
[20] Tagawa A, Kaneko T, Shinohara T, et al. Heme oxygenase-1 inhibits cigarette smoke-induced in the tracheal mucosal permeahility in guinea pigs in vivo[J]. Inflammation Research, 2005, 54(5): 229-234.