以SPF级Balb/c雌性小鼠为实验动物,对其适应性饲养3 d后连续30 d灌喂不同剂量L-茶氨酸,然后腹腔注射产肠毒性大肠杆菌E44813诱导免疫应激,5 h后取样,分析研究了各组小鼠肝脏与脾脏系数,血清谷丙转氨酶(Alanine aminotranferease, ALT)与谷草转氨酶(Aspartate aminotransferase, AST)含量,肝组织匀浆丙二醛(Malondialdehyde, MDA)水平与谷胱甘肽过氧化物酶(Glutathione peroxidase, GSH-PX)、过氧化氢酶(Catalase, CAT)、超氧化物歧化酶(Superoxide Dismutase, SOD)活性,肝组织病理学变化,以及血清中γ-干扰素(γ-Interferon, IFN-γ)、白细胞介素-4(Interleukin-4, IL-4)的表达量,以探明L-茶氨酸对产肠毒性大肠杆菌免疫应激小鼠肝脏的保护作用。结果表明,不同剂量的茶氨酸干预处理均能明显降低由大肠杆菌E44813感染引起的肝脏、脾脏系数的升高,降低血清ALT、AST及肝组织匀浆MDA水平,提高肝组织匀浆SOD、CAT、GSH-PX活性,减少IFN-γ、IL-4表达量,改善肝脏组织病理损伤,其中以300 mg·kg-1剂量组效果最好,说明茶氨酸干预对产肠毒性大肠杆菌诱导的免疫应激小鼠肝损伤具有保护作用,而且可能是通过减少IFN-γ与IL-4的表达、降低炎症反应、提高抗氧化能力等途径达到保护肝脏的作用。
SPF grade Balb/c female mice were used for this experiment. After adaptive feeding for 3 d, the mice were pre-treated orally with different dosages of L-theanine once daily for 30 d. ETEC E44813 intraperitoneal injection was subsequently performed and samples were collected 5h later. The coefficient of liver and spleen, levels of serum alanine aminotranferease (ALT), aspartate aminotransferase (AST) as well as liver homogenate malondialdehyde (MDA), and activities of liver homogenate superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), pathological changes of liver, expressions of serum γ-Interferon (IFN-γ) and Interleukin-4 (IL-4) were studied to explore the protective effects of L-theanine on liver of ETEC immune mouse model. Results show that pretreatment with L-theanine significantly reduced the increase of liver and spleen coefficient caused by ETEC infection|lowered the levels of serum ALT|AST and liver homogenate MDA|increased the activities of liver homogenate SOD|CAT|GSH-Px|decreased the expressions of serum IFN-γ|IL-4|and improved pathological injury of liver tissue. Therefore|it can be concluded that L-theanine is able to prevent ETEC-induced hepatic injury through depressing IFN-γ, IL-4 and inflammatory reaction and enhancing hepatocyte antioxidant abilities.
[1] 李中兴. 诊断细菌学[M]. 香港: 黄河文化出版社, 1992: 286-291.
[2] WHO IRIS.Future directions for research on enterotoxigenic Escherichia coli vaccines for developing countries[J]. Wkly Epidemiol Ree, 2006, 8l(11): 97-104.
[3] Qadri F, Svennerholm A M, Faruque A S, et al.Enterotoxigenic Escherichia coli in developing countries: epidemiology, microbiology, clinical features, treatment, and prevention[J]. Clin Micribiol Rev, 2005, 18(3): 465-483.
[4] 陈盼霖, 周明旭, 朱国强. 产肠毒素大肠杆菌研究进展和面临挑战[J]. 中国预防兽医学报, 2015, 37(10): 813-816.
[5] Qian Liu, Huiying Duan, Jinling Luan, et al.Effects of theanine on growth of human lung cancer and leukemia cells as well as migration and invasion of human lung cancer cells[J]. Cytotechnology, 2009, 59(3): 211-217.
[6] Zhang G, Miura Y, Yakasaki K.Effects of powdered green tea and theanine on tumor growth and endogenous hyperlipidemia in hepatoma-bearing rats[J]. Biosei Biotechnol Biochem, 2002, 66(4): 711-716.
[7] Sugiyama T, Sadzuka Y.Combination of theanine with doxorubicin inhibits hepatic metastasis of M5076 ovarian sarcoma[J]. Clinical Cancer Research, 1999, 5(5): 413-416.
[8] Sadzuka Y, Yamashita Y, Kishimoto S, et al.Glutamate transporter mediated increase of antitumor activity by theanine, an amino acid in green tea[J]. The Pharmaceutical Society of Japan, 2002, 122(11): 995-999.
[9] 雪玲, 程朝辉, 黄才欢, 等. 茶氨酸对小鼠学习记忆能力的影响[J]. 食品科学, 2004, 25(5): 171-173.
[10] 刘显明, 李月芬, 李国平. 茶氨酸对D-半乳糖衰老模型小鼠抗衰老作用的实验研究[J]. 刨伤外科杂志, 2008, 10(3): 257-259.
[11] Anne L Lardner.Neurobiological effects of the green tea constituent theanine and its potential role in the treatment of psychiatric and neurodegenerative disorders[J]. Nutritional Neuroscience, 2014, 17(4): 145-155.
[12] 彭彬, 刘仲华, 林勇, 等. L-茶氨酸改善慢性应激大鼠抑郁行为作用研究[J]. 茶叶科学, 2014, 34(4): 355-363.
[13] 王庆利, 张力军, 吴明春, 等. 茶氨酸对脑缺血损伤大鼠自由基代谢的影响[J]. 实用医学杂志, 2008, 24(11): 1898-1900.
[14] 王玉芬, 秦志祥, 唐振山, 等. 茶氨酸对家兔脑缺血后lL-8和NSE及脑超微结构的影响[J]. 中国医师杂志, 2007, 9(1): 4-6.
[15] 王小雪, 邱隽, 宋宇, 等. 茶氨酸的抗疲劳作用研究[J]. 中国公共卫生, 2002, 18(3): 315-318.
[16] Yokogoshi H, Mochizuki M, Saitoh K.Theanine-induced reduction of brain serotonin concentration in rats[J]. Bioscience Biotechnology Biochemistry, 1998, 62(4): 816-817.
[17] Li G, Ye Y, Kang J, et al.L-Theanine prevents alcoholic liver injury through enhancing the antioxidant capability of hepatocytes[J]. Food Chem Toxicol, 2012, 50(2): 363-372.
[18] 王宝恩, 张定凤. 现代肝脏病学[M]. 北京: 科学出版社, 2003, 67-77.
[19] Sakaguchi S, Furusawa S.Oxidative stress and septic shock: metabolic aspects of oxyfen-derived free radicals generated in the liver during endotoxemia[J]. FEMS Immunol Med Microbiol, 2009, 39(9): 167-177.
[20] 苏磊, 刘志锋. “肠-肝轴”与脓毒症[J]. 中华急诊医学杂志, 2010, 19(2): 124-125.