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1 7 6 Vol. 7 No Journal of Food Safety and Quality Jun., 米克热木沙衣布扎提 1, 陈琛 2, 王萌 3*, 王剑 2, 张小莺 (1., ; 2., ; 3., ) 3 摘要 : 目的 - 方法 42 6 :, -, (ALT) (AST) (H2O2) (MDA) (CAT) (T-SOD) (GSH) / CYP2E1 CYP2A5 结果 - CYP2E1 CYP2A5 ; CYP2E1 CYP2A5, H 2 O 2 MDA, CAT T-SOD GSH 结论 CYP2E1 CYP2A5 -, - 关键词 : ; ; P450 2E1; P450 2A5; Protection of pyrazole-lipopolysaccharide induced liver injury in mice by Prunella vulgaris L. MIKEREMU Shayi-buzhati 1, CHEN Chen 2, WANG Meng 3*, WANG Jian 2, ZHANG Xiao-ying 3 (1. College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi , China; 2. College of Biological Science and Engineering, Shaanxi University of Technology, Hanzhong , China 3. College of Veterinary Medicine, Northwest A&F University, Yangling , China) ABSTRACT: Objective To explore the protective effect of Prunella vulgaris L. (PV) against pyrazolelipopolysaccharide (LPS) induced liver injury in mice. Methods A total of 42 mice were randomly divided into 6 groups: normal control group, model group, positive control group and low, middle and high dose of PV group. Mice were treated with PV by oral gavage, and liver injury of model mice were treated with pyrazole-lps by the injection. The liver function was evaluated by detecting the activity of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and hepatic hydrogen peroxide (H 2 O 2 ), catalase (CAT), total super-oxidedismutase (T-SOD), glutathione (GSH), malondialdehyde (MDA), and the protein expression of CYP2E1 and CYP2A5. Results Pyrazole-LPS could induce liver injury by increase the CYP2E1 and CYP2A5 protein levels and oxidative stress. PV 基金项目 : ( ) 2011 ( ) Fund: Supported by the National Natural Science Foundation of China ( ), Construction Project of Shaanxi Collaborative Innovation Center (2011, Qinling-Bashan Mountains Bioresources Comprehensive Development) * 通讯作者 :,, Aplice415@163.com *Corresponding author: WANG Meng, Ph.D., College of Veterinary Medicine, Northwest A&F University, Yangling , China. Aplice415@163.com

2 6, : extraction could reduce the CYP2E1 and CYP2A5 protein levels, which led to the reduction of H 2 O 2 and MDA as well as the enhancement of anti-oxidative defense (CAT, T-SOD and GSH). Conclusion These findings showed the hepato-protective properties of PV against pyrazole-lps induced liver injury by attenuating the protein levels of CYP2E1 and CYP2A5, and enhancing the protective role of anti-oxidative defense system. KEY WORDS: Prunella vulgaris L.; liver injury; CYP2E1; CYP2A5; oxidative stress 1 引言 (Prunella vulgaris L.),, [1], [2], [3], [4] P450 (CYP450) I,, CYP2E1 CYP2A5 (pyrazole)- (lipopolysaccharide, LPS), [5], 2 材料与方法 2.1 材料 (Prunella vulgaris L., PV), 2015, ;,, (silybin, SL) Sigma (H 2 O 2 ) (MDA) (T-SOD) (GSH) (CAT) ;, 2.2 夏枯草水提物的制备 2 h, 30 min,, 1, 试验动物及分组, (20±2) g 42 6, 7,,, mg/kg,, 60 mg/kg, 2, 7 d, 4 :, 150 mg/kg, 2 /d, 2 d, 6 d LPS, 4 mg/kg, 2 /d, 24 h 2.4 样品采集与处理 24 h, 2 d, 3500 r/min 10 min,, (ALT) (AST),, 10%, (H 2 O 2 ) (MDA) (T-SOD) (GSH) (CAT) 2.5 Western blotting 检测细胞色素酶 2E1 和 2A5 的蛋白水平 [6], 3 SDS-PAGE, 5% 1 h, CYP2E1 2A5 (, ), (1 2000), DAB Gelpro32, P450 (CPR)(Abcam, ) 2.6 数据处理 SPSS 19.0 One Way ANOVA, LSD, ±, P<0.05, P> 结果与分析 3.1 夏枯草对肝脏生化指标水平的影响 - LPS 24 h ALT AST ( 1),

3 2336 7, -, - ALT AST, ALT AST, - - H 2 O 2 MDA (P<0.05), H 2 O 2 MDA ( 1) - - GSH CAT T-SOD (P<0.05), GSH CAT T-SOD ( 1) GSH CAT T-SOD,, - - CYP2E1, CYP2E1 (P<0.05) 1(B) 2, CPR CYP2A5, CYP2A5 56 Ku - CYP2A5, (P<0.05), 2, -, CYP2E1 (P<0.05), (P>0.05), - CYP2A5 3.2 夏枯草对肝脏 CYP2E1 和 CYP2A5 蛋白水平的影响 CPR CYP2E1, 77 Ku, CYP2E1 57 Ku - CYP2E1, (P<0.05), 1(A) 2, 1 CYP2E1 CYP2A5 Fig. 1 Effects of Prunella vulgaris L. on CYP2E1 and CYP2A5 protein levels 表 1 夏枯草对血清 ASL 和 AST 活性的影响 (n=7) Table 1 Effects of Prunella vulgaris L. on serum ALT and AST activities (n=7) - ALT (U/L) d a b c d c AST (U/L) c a b b c c H 2 O 2 (mmol/ml) d a b b c c MDA (nmol/ml) c a a b b b T-SOD (U/mL) a c b b a a GSH (µmol/l) b c b a a b CAT (U/g) a b b a a a : ±, (P<0.05); (P>0.05) 表 2 夏枯草对 CYP2E1 和 CYP2A5 的蛋白表达水平的影响 (n=7) Table 2 Effects of Prunella vulgaris L. on CYP2E1 and CYP2A5 protein levels (n=7) - CYP2E d a b c d c CYP2A c a b b c c : ±, (P<0.05); (P>0.05)

4 6, : 讨论 CYP2E1 CYP2A5,, [8-10] CYP2E1 CYP2A5, 24 h CYP2E1, LPS [10],,, (H 2 O 2 ),, AST,, ALT AST, ALT AST,, MDA, GSH, CAT SOD, CYP2E1, CYP2E1 [12] 2~3 d, CYP2E1 2~4 [13], - CYP2E1 3 ( 2), - CYP2E1 CYP2E1, [13] CYP2E1 ROS, H 2 O 2 LPS [14],, LPS [15] CYP2E1, CYP2A5 [9] CYP2A5,, - H 2 O 2 8, T-SOD, CAT GSH /, -,, [16] [17,18] [19], Wang [6] Tung [20], -,, CYP2E1 CYP2A5, CYP2E1 CYP2A5,, 1, H 2 O 2 MDA, T-SOD CAT GSH / SOD, 2, H 2 O 2 O - 2 ; CAT H 2 O 2 O - 2 [21] ; GSH [22] -, 5 小结 CYP2E1 CYP2A5 - - CYP2E1 CYP2A5,, -, CYP2E1 CYP2A5, H 2 O 2 MDA ;, T-SOD CAT GSH, 参考文献 [1]. [J]., 2015, 12(31): 57. Zhang SN. Sefheal medicinal composition and efficacy were reviewed [J]. Beijing Agric, 2015, 12(31): 57. [2],,,. [J]., 2003, 20(1): Liu Y, Song SJ, Xu SX, et al. Advances in the study on the chemical constituents and biological activities of Prunella vulgaris L. [J]. J Shenyang Pharm Univ, 2003, 20(1): [3],,,. [J]., 2011, 17(6): Feng RX, Zhang SH, Lin C, et al. Study on compound sefheal protect liver enzyme [J]. Chin J Exp Tradit Med Form, 2011, 17(6): [4] Li S, Tan HY, Wang N, et al. The role of oxidative stress and antioxidants in liver diseases [J]. Int J Mol Sci, 2015, 16(11): [5] Uhl P, Fricker G, Haberkorn U, et al. Current status in the therapy of liver diseases [J]. Int J Mol Sci, 2014, 15(5): [6] Wang M, Sun J, Jiang Z, et al. Hepatoprotective effect of kaempferol against alcoholic liver injury in mice [J]. Am J Chin Med, 2015, 43(2): [7],,,. CYP2E1 [J]., 2014, 5(6): Wang T, Wang M, Jiang ZH, et al. Effect of flavonoids extract from apocynum venetum tea on CYP2E1 in mice [J]. J Food Saf Qual, 2014, 5(6): [8] Juvonen RO, Kaipainen PK, Lang MA, et al. Selective induction of coumarin 7-hydroxylase by pyrazole in D2 mice [J]. Eur J Biochem, 1985, 152(1): 3 8. [9] Gilmore WJ, Hartmann G, Piquette-Miller M, et al. Effects of lipopolysaccharide-stimulated inflammation and pyrazole-mediated

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5 hepatocellular injury on mouse hepatic CYP2A5 expression [J]. Toxicology, 2003, 184(2): [10] Lang MA, Juvonen R, Järvinen P, et al. Mouse liver P450Coh: genetic regulation of the pyrazole-inducible enzyme and comparison with other P450 isoenzymes [J]. Arch Biochem Biophys, 1989, 271(1): [11] Lu Y, Gong P, Cederbaum AI, et al. Pyrazole induced oxidative liver injury independent of CYP2E1/2A5 induction due to Nrf 2 deficiency [J]. Toxicology, 2008, 252(1): [12] Wu D, Cederbaum AI. Ethanol and arachidonic acid produce toxicity in hepatocytes from pyrazole-treated rats with high levels of CYP2E1 [J]. Mol Cell Biochem, 2000, 204(1 2): [13] Winters DK, Cederbaum AI. Time course characterization of the induction of cytochrome P-450 2E1 by pyrazole and 4-methylpyrazole [J]. Biochim Biophys Acta, 1992, 1117(1): [14],,,. [J]., 2015, 42 (1): Yang Z, Gao M, Xiao K, et al. Effect of visfatin on morphological structure and MPO in Liver of LPS-induced Rats [J]. China Anim Husb Vet Med, 2015, 42(1): Tao N. Selfheal alcohol extract from the protection of acute liver injury in rats [J]. Asia-Pacific Tradit Med, 2015, 11(15): [19],,,. [J]. ( ), 2016, (2): 23 27, 34. Chen T, Su YH, Huang YL, et al. Protective effects of aqueous extract of Prunella vulgaris L. on hepatic injured mice [J]. J Northwest A&F Univ (Nat Sci Ed), 2016, (2): 23 27, 34. [20] Tung YT, Wu JH, Huang CC, et al. Protective effect of Acacia confusa bark extract and its active compound gallic acid against carbon tetrachloride-induced chronic liver injury in rats [J]. Food Chem Toxicol, 2009; 47 (6): [21] Vuleta A, Jovanović SM, Tucić B, et al. Adaptive flexibility of enzymatic antioxidants SOD, APX and CAT to high light stress: the clonal perennial monocot Iris pumila as a study case [J]. Plant Physiol Biochem, [22] Wang M, Sun J, Jiang Z, et al. Hepatoprotective effect of kaempferol against alcoholic liver injury in mice [J]. Am J Chin Med, 2015, 43(2): ( 责任编辑 : 金延秋 ) [15] Lu Y, Cederbaum AI. Enhancement by pyrazole of lipopolysaccharideinduced liver injury in mice: role of cytochrome P450 2E1 and 2A5 [J]. Hepatology, 2006, 44(1): 作者简介 [16],. [J]., 2014, 29(3): Cui TS, Miao MS. Chemistry, pharmacology and clinical applications of selfheal [J]. Chin Med, 2014, 29(3): 米克热木沙衣布扎提, 硕士, 副教授, 硕士生导师, 主要研究方向为基础兽医 @qq.com [17],. [J]., 2013, 31(2): Chen T, Chen RG. Protection effect of Prunella Vulgaris L. on carbon tetrachloride induced liver in mice [J]. J Longyan Univ, 2013, 31(2): 王萌, 博士, 主要研究方向为新兽药研发 Aplice415@163.com [18]. [J]., 2015, 11(15):

中草药 Chinese Traditional and Herbal Drugs 第 46 卷第 9 期 2015 年 5 月 1339 非酒精性脂肪性肝病 (nonalcoholic fatty liver disease,nafld) 是一种与胰岛素抵抗遗

中草药 Chinese Traditional and Herbal Drugs 第 46 卷第 9 期 2015 年 5 月 1339 非酒精性脂肪性肝病 (nonalcoholic fatty liver disease,nafld) 是一种与胰岛素抵抗遗 1338 中草药 Chinese Traditional and Herbal Drugs 第 46 卷第 9 期 2015 年 5 月九味肝泰胶囊对高脂饮食诱导大鼠非酒精性脂肪肝的治疗作用陈菲 1 2,, 艾国 3, 盛柳青 1, 麻佳蕾 1, 黄正明 1. 金华职业技术学院医学院, 浙江金华