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1 6 9 Vol. 6 No Journal of Food Safety and Quality Sep., , 陈金华 2 1,, 谭斌 2, 黄建安 1*, 刘仲华 (1., ; 2., ) 1, 2 摘要 : 目的方法, (black tea, BT), (malondialdehyde, MDA) (superoxide dismutase, SOD) (glutathione peroxidase, GSH-Px) (alanine transaminase, ALT) (aspartate transaminase, AST), 结果, ALT AST MDA, SOD GSH-Px, 结论, 关键词 : ; ; ; Protective effects of black tea on mice liver under high fat diet CHEN Jin-Hua 1, 2, TAN Bin 1, 2, HUANG Jian-An 1*, LIU Zhong-Hua 1, 2 (1. Key Lab of Tea Science of Ministry of Education, Hunan Agricultural University, Changsha , China; 2. National Research Center of Engineering & Technology for Utilization of Functional Ingredients from Botanicals, Hunan Agricultural University, Changsha , China) ABSTRACT: Objective To study the protective effect of black tea (BT) on mice liver under a high fat diet. Methods The mice were fed with BT of different types and quantities under the condition of high fat diet. Liver index and content of liver malondialdehyde (MDA), liver superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), serum glutamic-pyruvic transaminase (ALT) and glutamic-oxalacetic transaminase (AST) were measured, and the histopathological changes in liver tissues were observed with unaided eyes and under an optical microscope as well. Results The content of serum ALT, AST, liver MDA and liver index in the black tea feeding group were lower than the high hyperlipoidemia model group, while liver SOD activity and GSH-Px activity were higher, and the liver lesions were reduced. Conclusion It indicates that BT has positive effect on protecting and repairing mice liver with high fat diet. The mechanisms may be that black tea has effects on ameliorating oxidative stress and reducing lipid peroxidation product. KEY WORDS: black tea; high fat diet; mice; liver 基金项目 : (CARS-23-C) (2013FJ4222) (13C421) Fund: Supported by the National Tea Industry Technology System Project (CARS-23-C), the Provincial Science and Technology Projects (2013FJ4222) and the Scientific Research Fund of Hunan Provincial Education Department (13C421) * 通讯作者 :,, jian 7513@sina.com *Corresponding author: HUANG Jian-An, Professor, Key Lab of Tea Science of Ministry of Education, Hunan Agricultural University, Changsha , China. jian 7513@sina.com

AST MDA, SOD GSH-Px, 结论, 关键词 : ; ; ; Protective effects of black tea on mice liver under high fat diet CHEN Jin-Hua 1, 2, TAN Bin 1, 2, HUANG Jian-An 1*, LIU Zhong-Hua 1, 2 (1.

2 引言,,,,,,,,,,,, [1],,,,, [2-8],,, [9-15] (superoxide dismutase, SOD) (glutathione peroxidase, GSH-Px),, ; (malondialdehyde, MDA),, (alanine transaminase, ALT) (aspartate transaminase, AST),,,,,,, ;,,, MDA SOD GSH-Px ALT AST,, 2 材料与方法 2.1 材料与设备主要仪器 (TBA-40FR, ); (EG 1150C, Leica ); (TP1020, Leica ); (RM2235, Leica ); (DM2000, Leica ); (MIKRO 22R, Hettich ); (Modulyod-230, ); (BUCHI R-200, BUCHI ); (F6/10, FLUKO); (Thermo Scientific Varioskan Flash, Thermo ); (Eppdrof) 主要试剂 ( ALT) (AST), ; (MDA) (SOD) (GSH-Px), 实验动物及饲养条件 SPF ICR, 30 g, 5,, : SCXK( ) : 20~25, 45%~55%,,,, 1 d 1, 供试材料 (2011, ), (2011, ), (2011, ), (2011, ), (2012, ), (2011, ), (2011, ) 动物饲料, : SCXK( ) , : 24.3% 7.5% 8.6% 3.5% 0.7% 1.16% 7.5% [16] :, 78.8%;, 1%;, 10%;, 10%;, 0.2%

3 9, : 阳性药 : (, ) 2.2 实验方法红茶粉的制备, 1:10(m:V) (100 ), 30 min, ; 1:5 20 min, 10 min,,,, h 24 h,,, 动物实验 (1) ( 60 kg 10 g, mg kg -1 d -1 ) 5 10( ) 20 [17] (60 kg) 10 mg kg -1 d -1, 9.1 [18], 91 mg kg -1 d ml/10 g bw [19] (2) 3 d, 7 ( ), 24 10,,,,, 1, 35 d [17,19] 2.3 指标测定血清 ALT AST 的测定 12 h( ),, r/min 10 min,, ALT AST 肝匀浆制备及 MDA SOD GSH-PX 测定,,,, 0.2 g,, 10%, r/min 10 min, -20 MDA SOD GSH-PX 小鼠肝脏的组织病理切片制备及观察 0.5 g, 10%, ; HE 2.4 数据处理 Excel 2010 SPSS 19.0, ± ( x ±s), (least significant difference, LSD), P 结果与分析 3.1 红茶对小鼠肝体比的影响 1,,,,,, (P 0.01),, 3.2 红茶对小鼠 MDA SOD 和 GSH-Px 的影响 SOD GSH-Px MDA 2 3 4, MDA, SOD GSH-Px,, SOD GSH-Px (P 0.01), MDA, (P 0.01), 3.3 红茶对小鼠血清 ALT 和 AST 的影响 ALT AST 1, ALT AST,,

3.2 肝匀浆制备及 MDA SOD GSH-PX 测定,,,, 0.2 g,, 10%, 4 3000 r/min 10 min, -20 MDA SOD GSH-PX 2.3.3 小鼠肝脏的组织病理切片制备及观察 0.5 g, 10%, ; HE 2.4 数据处理 Excel 2010 SPSS 19.

4 3640 食品安全质量检测学报 Fig. 1 图 1 红茶对小鼠肝指数的影响 Effects of black tea (BT) on hepatic index of mice 注: CK: 正常对照组, M: 模型对照组, Y: 阳性药对照组与模型对照组比较* P<0.05, ** P<0.01 图与此相同图 2 红茶对小鼠肝匀浆 SOD 的影响 Fig. 2 Effects of BT on liver SOD of mice 图 3 红茶对小鼠肝匀浆 GSH-Px 的影响 Fig. 3 Effects of BT on liver GSH-Px of mice 第6卷

5 9, : MDA Fig. 4 Effects of BT on liver MDA of mice,, ALT AST, (P 0.05) (P 0.01), 1 ALT AST,, ALT AST 3.4 红茶对小鼠肝组织病理形态学的影响小鼠肝脏肉眼观察结果, ( 5),,,,, ;,,,, ;,,,,,,,,,,, 5,, :,, ;, 表 1 红茶对小鼠血清 ALT AST 水平的影响 ( x±s) Table 1 Effects of BT on ALT and AST in serum of mice n ALT (U/L) AST (U/L) ± ± ± ± ± ± ±9.83 * ±54.46 ** ±8.62 * ±46.47 * ±15.57 * ±61.34 * ±9.67 ** ±55.89 ** ±9.73 * ±59.71 ** ± ±54.62 ** ± ±45.59 * ±9.34 ** ±42.57 ** ±13.23 ** ±67.85 ** ±12.99 * ±36.48 ** ±10.36 ** 99.13±51.31 ** ±7.09 ** 91.14±29.61 ** ± ± ±14.56 ** ±74.64 ** ± ± ±6.63 ** 91.50±32.89 ** ± ±41.97 ** ±11.89 * ±49.72 * ± ±43.52 * ±12.05 ** 96.14±59.57 ** ±6.63 ** 82.67±31.60 ** : * P<0.05, ** P<0.01

89 ** 10 32.00±9.73 * 122.86±59.71 ** 10 34.40±14.69 119.80±54.62 ** 10 40.13±15.06 143.88±45.59 * 10 29.13±9.34 ** 122.13±42.57 ** 10 29.14±13.23 ** 118.14±67.85 ** 10 33.25±12.99 * 109.13±36.

6 3642 第6卷食品安全质量检测学报小鼠肝组织显微镜观察结果小鼠肝组织 HE 染色后显微镜观察并拍照, 结果如图 6 所示正常对照组小鼠肝脏组织结构完整, 肝小叶轮廓中央静脉肝细胞索及肝窦清晰, 肝细胞索围绕中央静脉呈放射状排列, 肝细胞大小较一致, 细胞核清晰, 胞浆均匀(图 6-CK) 而饲喂 5 周高脂饲料的模型组, 肝细胞组织结构异常, 肝小叶结构紊乱且界限模糊, 肝窦变窄甚至消失, 肝细胞排列辐射状模糊, 肝小叶周围肝细胞胞浆出现大量的圆形空泡, 造成严重脂肪变性(图 6-M) 相比于高脂模型组, 进食高脂饲料同时灌喂低中高剂量红茶能够梯度性的改善由高脂饲料引起的肝组织病变(图 6), 肝组织切片观察显示, 低剂量组肝组织有严重水样变, 中剂量组轻微水样变, 但均未形成脂肪空泡, 高剂量组肝组织基本恢复正常, 与阳性药效果相当, 肝小叶周围的肝细胞发生水泡的细胞数量减少, 脂滴明显减少甚至消失, 肝细胞肿胀变性的程度有所降低, 细胞间界限较清晰说明红茶对高脂饮食小鼠肝脂肪变性具有较强的修复作用, 有效减轻了小鼠的肝损伤 4 讨论研究结果提示, 高脂饮食可造成小鼠体内严重的氧化与抗氧化的失衡, 使机体肝组织正常的抗氧化能力明显受到损伤而红茶能显著升高肝脏 SOD 与 GSH-Px 酶的活性, 与此同时还可以降低 MDA 的含量,提高肝细胞的抗氧化能力, 有效清除机体自由基, 调节和改善自由基代谢平衡, 改善肝脏脂质过氧化状态, 减少脂质过氧化产生的毒素, 从而达到保护肝脏以及修复已损伤的肝组织的功能有研究表明, 茶多酚茶黄素及茶多糖具有清除氧自由基和抗脂质过氧化的能力 [20-22], 红茶在体内对肝组织氧化损伤保护作用的机制很可能是与其有关一方面红茶 Fig. 5 图 5 小鼠肝照片 Liver photos of mice in different groups 注: CK-空白对照组; M-高脂模型对照组; Y-阳性药组; JD-将军红低剂量组; JZ-将军红中剂量组; JG-将军红高剂量组; SD-石门红茶中的茶多酚与茶黄素含有酚性羟基, 为清除自由基团提供了位点, 易氧化成醌类而提供 H+, H+可结合自由基而将其还原为惰性化合物, 防止损伤肝脏; 红低剂量组; SZ-石门红茶中剂量组; SG-石门红茶高剂量组; QD- 另一方面, 红茶中含有较高的茶多糖, 该类物质具祁门红茶低剂量组; QZ-祁门红茶中剂量组; QG-祁门红茶高剂量有较好的抗氧化作用, 能清除多种自由基, 以防其组; ND-宁红低剂量组; NZ-宁红中剂量组; NG-宁红高剂量组; FD- 对肝脏的损伤凤庆红茶低剂量组; FZ-凤庆红茶中剂量组; FG-凤庆红茶高剂量组; DD-大唐英红低剂量组; DZ-大唐英红中剂量组; DG-大唐英红本研究发现, 高脂模型组小鼠血清 ALT 与 AST 高剂量组; XD-霞山红低剂量组; XZ-霞山红中剂量组; XG-霞山红活力均有所升高, 说明高脂饮食对肝脏正常功能造高剂量组图 6 中相同成了一定的影响而红茶各剂量组均不同程度地降低

性药效果相当, 肝小叶周围的肝细胞发生水泡的细胞数量减少, 脂滴明显减少甚至消失, 肝细胞肿胀变性的程度有所降低, 细胞间界限较清晰说明红茶对高脂饮食小鼠肝脂肪变性具有较强的修复作用, 有效减轻了小鼠的肝损伤 4 讨论研究结果提示, 高脂饮食可造成小鼠体内严重的氧化与抗氧化的失衡, 使机体肝组织正常的抗氧化能力明显受到损伤而红茶能显著升高肝脏 SOD 与 GSH-Px

7 第9期陈金华, 等: 红茶对高脂饮食小鼠肝脏的保护作用研究 Fig 图 6 小鼠肝组织形态学变化( 100) Changes of morphology of liver tissues in mice ( 100) 血清中 ALT 与 AST 活力, 这提示红茶能够保护并修小鼠喂食高脂饲料后, 肝组织肉眼观察与肝脏复和稳定肝细胞的质膜线粒体膜的作用与模型对切片显微镜观察显示, 肝细胞及肝小叶结构出现了照组比较, 灌胃各剂量红茶的小鼠肝脏指数显著降异常, 而同时灌胃了不同剂量红茶后发现, 胞浆内低, 这在一定程度上与红茶中一些酚类物质及其氧脂滴减少或消失, 无明显炎细胞坏死或浸润, 其中化产物的抗氧化以及抗炎作用有关 [20-22] 效地减轻了肝脏的水肿和病变症状, 从而也有高剂量红茶组更接近正常对照组肝脏情况, 而低剂量组相比高脂模型组, 肝组织病变程度有所改善,

这提示红茶能够保护并修小鼠喂食高脂饲料后, 肝组织肉眼观察与肝脏复和稳定肝细胞的质膜线粒体膜的作用与模型对切片显微镜观察显示, 肝细胞及肝小叶结构出现了照组比较,

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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