牛黄参胶囊对小鼠免疫性肝损伤保护作用的实验研究
摘要
1.目的
本实验针对病毒性肝炎发病的免疫学机制,采用卡介苗(BCG)加酯多糖(LPS)联合诱导的免疫性肝损伤模型,通过测定动物血清ALT. AST值、肝组织超氧化物歧化酶(SOD)活力、脂质过氧化物丙二醛(MDA)的含量、肝组织病理形态及超微结构、过氧化损伤及免疫调节等途经,并做统计分析,评价牛黄参胶囊对免疫性肝损伤保护作用的效果,并探讨其作用机制,为牛黄参胶囊抗免疫性肝损伤提供了科学依据,并为牛黄参胶囊的进一步开发应用提供依据。
2.方法
将60只昆明小鼠随机分为正常组、BCG加LPS模型组、联苯双酯组、牛黄参胶囊高、中、低剂量组,每组1 0只。具体方法：按体重将60只小鼠编为1——60号,用DPS7.55统计软件进行处理：试验设计→完全随机分组→实验样本数60,分组组数6→确定,随机分为6组。
实验室喂养一周,参考文献方法造模,除正常对照组外,其余组第一天均尾静脉注射BCG2.5mg/0.2m1.只(约5×107个活菌)造模。正常对照组、BCG加LPS模型组,自来水0.5ml/只,ig10天；联苯双酯组剂量12mgkg,ig10天。牛黄参胶囊高、中、低剂量组分别给予牛黄参胶囊内容物水溶液以3400mg/kg.1700mgkg.850mg/kg灌胃10天。除正常对照组外,各组动物均于末次给药后,尾静脉注射LPS7.5μg/0.2ml.只。具体方法：先将小鼠固定于小鼠固定器内,让尾部全部露到固定器外,用75%乙醇擦拭尾部,使其充血。选择一根最为充盈的血管,用左手捏住尾巴的末端,右手持装有四号针头的注射器,以30°角进行静脉穿刺推注药液无阻力感,且可见沿静脉血管出现一条白线,则表明针头确实在血管内,即可继续推完药液。注射完毕后,拔出针头,轻按注射部位止血。注射后禁食不禁水,10小时后,进行模型处理。小鼠用0.03g/10g水合氯醛麻醉后,眼眶取血1.0ml,以无菌试管采集,离心血清,上全自动血生化仪,检测ALT、AST水平。采血前称小鼠体重,采血后断头处死小鼠,小鼠处死后取肝组织,用20%甲醛固定,常规石蜡包埋切片,行HE染色,光镜观察组织病理,参照免疫性肝损伤实验研究相关文献和肝组织形态学相关研究文献资料,根据肝组织炎症、坏死及出血程度,将肝组织病理改变分级。
切取左前叶肝组织0.5—1.0 g,冷生理盐水冲洗,滤纸吸干,称重,冰浴中制成10%肝匀浆,以4000转/min,低温离心15分钟,取上清液。按步骤测定SOD活性,检测丙二醛含量,其操作步骤按试剂盒说明书进行。TNF-α活性、IL-6活性检测采用ELISA法,按试剂盒说明操作。实验数据用均数±标准差(X士S)表示,组内比较：t检验；组间比较：方差分析。P<0.05认为差异具有统计学意义。用SPSS 13.0统计软件分析。
3.结果
(1)各治疗组ALT、AST值较模型组明显降低(P<0.05),说明牛黄参胶囊能有效防止免疫性肝损伤引起的肝功能减退。
(2)各治疗组病理结构明显优于模型组(P<0.05),证明牛黄参胶囊能有效防止BCG加LPS对小鼠肝脏组织结构的免疫性损伤。
(3)各治疗组肝组织匀浆SOD活性明显高于模型组,MDA含量明显低于模型组,差异有显著性意义(P<0.05),说明牛黄参胶囊具有抗氧化、清除自由基的作用。
(4)各治疗组TNF-α、IL-6的含量明显低于模型组,差异有显著性意义(P<0.05),说明牛黄参胶囊抗肝损伤的作用可能与其抑制细胞毒因子TNF-α、IL-6的释放有关。
(5)中剂量组牛黄参胶囊的各项指标与联苯双酯组比较无显著性差异,说明其疗效与联苯双酯相当。
4.结论
(1)牛黄参胶囊能减轻BCG加LPS免疫性肝损伤对小鼠肝脏病理结构的破坏,保护其肝功能。
(2)牛黄参胶囊最佳剂量与联苯双酯疗效相当。
1.Purpose
This experiment for viral hepatitis, using the pathogenesis of immunological mechanisms BCG plus LPS polysaccharides joint induction reexposed liver damage model, by means of measuring the animal serum ALT, AST value, liver tissue superoxide dismutase (SOD) vigor, lipid peroxide malondialdehyde (MDA), liver tissue pathology form and ultrastructure, peroxidation damage and immune adjusting etc through, and do statistical analysis, evaluation NHS on immunity liver injury protection effect, and discuss the mechanism for NHS capsule resistance to damage the liver immunity provides scientific basis for NHS and participation in clinical application and development of capsule to provide the basis.
2.Methods
Kunming mice only 60 were randomly divided into normal group, BCG add LPS model group, Biphenyl Dicarboxylate group, NHS capsule of high,medium and low doses groups of 10 only. Methods:according to the weight will be 60 mice dungenzhai yishu 1-60#, with DPS7.55 statistical software for processing:experimental design and the complete randomized-60, grouping experiments sample number of groups 6, OK.
Laboratory feeding a week, reference method made modules, except normal control group outside, the rest group first day all tail intravenous BCG2.5mg/0.2ml. one (about 5×107 living bacterium) made moulds. Normal control group, BCG add LPS model group,tap water 0.5ml. ig 10 days; only, Biphenyl Dicarboxylate group 12mg/kg doses, ig 10 days. NHS capsule is tall, medium, low doses groups were given NHS 3400mg/kg,1700mg/kg,850mg/kg gastric lavage 10 days. Besides the normal control group outside, each animal in all the end time after the treatment, tail intravenous LPS 7.5μg/0.2 ml one. Methods:first mouse fixed in mice holder, let the tail to set all dew outside, with 75% ethanol is wiped the tail, make its hyperemia. Choose a root most plentiful vein, left-handed means and index finger knead tail end, right hand holding the syringe needle with 4, with 30 degree Angle were venipuncture push note solution frictionless feeling, and visible along the veins appear a white line, indicates that needle indeed inside blood-vessel, can continue to push finish solution. After completion, uproot needle injection, lightly in injection site bleeding. After injection fasting not water, after 10 hours for model processing.Mice with 0.03g/10g hydration after anesthesia, orbit chlorine aldehyde take blood 1.0 ml, with sterile tubes collection, centrifugal serum, blood biochemistry analyzer, the automatic detection ALT, AST level. Former says mice weight, blood collection after death mice, anatomical dislocated take liver and spleen weighing, calculation, liver and spleen visceral index. Mice executed after take liver tissue, using formaldehyde is fixed, the conventional 20% paraffin embedding slice, did HE histopathological staining light microscopy, consult immunity liver injury experimental study relevant literatures and liver tissue morphological related research literature, according to the liver tissue inflammation and necrosis and haemorrhage degree, will the liver tissue pathology change five levels.
Cut take left medial liver tissue 0.5-1.0g, cold saline flush filter paper blot, weighing, ice bath of liver homogenate made by 10%,4000 turn/min, cryogenic centrifugal 15min, fetch the supernatant fluid. According to the instruction using glucosinolate and detect malondialdehyde (mda), determining SOD activity. Its operation step by kit instructions. TNF-cactivity, IL-6 active detection by ELISA, press kit that operation. Experimental data with mean+standard deviations (X±S) said, and in groups; t test comparison:Comparison:analysis of variance between groups. P<0.05 think differences are statistically significant. With SPSS 13.0 statistical software analysis.
3.Results
(1) The treatment group ALT, AST value significantly lower than that in model group(P<0.05),indicating that NHS capsules can effectively prevent the immunity liver damage caused by the liver function decreases.
(2) The treatment group pathological structure obviously better than the model group(P<0.05), proof NHS capsules can effectively prevent the BCG add LPS of mouse liver tissue structure reexposed damage.
(3) Various treatment group liver tissue homogenate SOD was obviously higher than model group, MDA content obviously lower than model group,there is a significant difference significance(P<0.05),indicating that NHS capsule of antioxidation, scavenging free radicals role.
(4) Each treatment group TNF-α;IL-6 value obviously lower than model group,there is a significant difference significance(P<0.05), indicating that NHS capsule resistant to damage the liver function probably with its inhibition cytotoxic factor TNF-α,IL-6 the release of the relevant.
(5) Medium NHS capsule group all indexes of double ester group compared with biphenyl non-significance difference, explain various aspects and Biphenyl Dicarboxylate curative effect quite.
4.Conclusion
(1) NHS capsules can reduce BCG and LPS immunity liver damage on mice hepatic pathology structural damage and to protect its liver function.
(2) NHS capsule dose and Biphenyl Dicarboxylate curative effect quite.
本实验针对病毒性肝炎发病的免疫学机制,采用卡介苗(BCG)加酯多糖(LPS)联合诱导的免疫性肝损伤模型,通过测定动物血清ALT. AST值、肝组织超氧化物歧化酶(SOD)活力、脂质过氧化物丙二醛(MDA)的含量、肝组织病理形态及超微结构、过氧化损伤及免疫调节等途经,并做统计分析,评价牛黄参胶囊对免疫性肝损伤保护作用的效果,并探讨其作用机制,为牛黄参胶囊抗免疫性肝损伤提供了科学依据,并为牛黄参胶囊的进一步开发应用提供依据。
2.方法
将60只昆明小鼠随机分为正常组、BCG加LPS模型组、联苯双酯组、牛黄参胶囊高、中、低剂量组,每组1 0只。具体方法：按体重将60只小鼠编为1——60号,用DPS7.55统计软件进行处理：试验设计→完全随机分组→实验样本数60,分组组数6→确定,随机分为6组。
实验室喂养一周,参考文献方法造模,除正常对照组外,其余组第一天均尾静脉注射BCG2.5mg/0.2m1.只(约5×107个活菌)造模。正常对照组、BCG加LPS模型组,自来水0.5ml/只,ig10天；联苯双酯组剂量12mgkg,ig10天。牛黄参胶囊高、中、低剂量组分别给予牛黄参胶囊内容物水溶液以3400mg/kg.1700mgkg.850mg/kg灌胃10天。除正常对照组外,各组动物均于末次给药后,尾静脉注射LPS7.5μg/0.2ml.只。具体方法：先将小鼠固定于小鼠固定器内,让尾部全部露到固定器外,用75%乙醇擦拭尾部,使其充血。选择一根最为充盈的血管,用左手捏住尾巴的末端,右手持装有四号针头的注射器,以30°角进行静脉穿刺推注药液无阻力感,且可见沿静脉血管出现一条白线,则表明针头确实在血管内,即可继续推完药液。注射完毕后,拔出针头,轻按注射部位止血。注射后禁食不禁水,10小时后,进行模型处理。小鼠用0.03g/10g水合氯醛麻醉后,眼眶取血1.0ml,以无菌试管采集,离心血清,上全自动血生化仪,检测ALT、AST水平。采血前称小鼠体重,采血后断头处死小鼠,小鼠处死后取肝组织,用20%甲醛固定,常规石蜡包埋切片,行HE染色,光镜观察组织病理,参照免疫性肝损伤实验研究相关文献和肝组织形态学相关研究文献资料,根据肝组织炎症、坏死及出血程度,将肝组织病理改变分级。
切取左前叶肝组织0.5—1.0 g,冷生理盐水冲洗,滤纸吸干,称重,冰浴中制成10%肝匀浆,以4000转/min,低温离心15分钟,取上清液。按步骤测定SOD活性,检测丙二醛含量,其操作步骤按试剂盒说明书进行。TNF-α活性、IL-6活性检测采用ELISA法,按试剂盒说明操作。实验数据用均数±标准差(X士S)表示,组内比较：t检验；组间比较：方差分析。P<0.05认为差异具有统计学意义。用SPSS 13.0统计软件分析。
3.结果
(1)各治疗组ALT、AST值较模型组明显降低(P<0.05),说明牛黄参胶囊能有效防止免疫性肝损伤引起的肝功能减退。
(2)各治疗组病理结构明显优于模型组(P<0.05),证明牛黄参胶囊能有效防止BCG加LPS对小鼠肝脏组织结构的免疫性损伤。
(3)各治疗组肝组织匀浆SOD活性明显高于模型组,MDA含量明显低于模型组,差异有显著性意义(P<0.05),说明牛黄参胶囊具有抗氧化、清除自由基的作用。
(4)各治疗组TNF-α、IL-6的含量明显低于模型组,差异有显著性意义(P<0.05),说明牛黄参胶囊抗肝损伤的作用可能与其抑制细胞毒因子TNF-α、IL-6的释放有关。
(5)中剂量组牛黄参胶囊的各项指标与联苯双酯组比较无显著性差异,说明其疗效与联苯双酯相当。
4.结论
(1)牛黄参胶囊能减轻BCG加LPS免疫性肝损伤对小鼠肝脏病理结构的破坏,保护其肝功能。
(2)牛黄参胶囊最佳剂量与联苯双酯疗效相当。
1.Purpose
This experiment for viral hepatitis, using the pathogenesis of immunological mechanisms BCG plus LPS polysaccharides joint induction reexposed liver damage model, by means of measuring the animal serum ALT, AST value, liver tissue superoxide dismutase (SOD) vigor, lipid peroxide malondialdehyde (MDA), liver tissue pathology form and ultrastructure, peroxidation damage and immune adjusting etc through, and do statistical analysis, evaluation NHS on immunity liver injury protection effect, and discuss the mechanism for NHS capsule resistance to damage the liver immunity provides scientific basis for NHS and participation in clinical application and development of capsule to provide the basis.
2.Methods
Kunming mice only 60 were randomly divided into normal group, BCG add LPS model group, Biphenyl Dicarboxylate group, NHS capsule of high,medium and low doses groups of 10 only. Methods:according to the weight will be 60 mice dungenzhai yishu 1-60#, with DPS7.55 statistical software for processing:experimental design and the complete randomized-60, grouping experiments sample number of groups 6, OK.
Laboratory feeding a week, reference method made modules, except normal control group outside, the rest group first day all tail intravenous BCG2.5mg/0.2ml. one (about 5×107 living bacterium) made moulds. Normal control group, BCG add LPS model group,tap water 0.5ml. ig 10 days; only, Biphenyl Dicarboxylate group 12mg/kg doses, ig 10 days. NHS capsule is tall, medium, low doses groups were given NHS 3400mg/kg,1700mg/kg,850mg/kg gastric lavage 10 days. Besides the normal control group outside, each animal in all the end time after the treatment, tail intravenous LPS 7.5μg/0.2 ml one. Methods:first mouse fixed in mice holder, let the tail to set all dew outside, with 75% ethanol is wiped the tail, make its hyperemia. Choose a root most plentiful vein, left-handed means and index finger knead tail end, right hand holding the syringe needle with 4, with 30 degree Angle were venipuncture push note solution frictionless feeling, and visible along the veins appear a white line, indicates that needle indeed inside blood-vessel, can continue to push finish solution. After completion, uproot needle injection, lightly in injection site bleeding. After injection fasting not water, after 10 hours for model processing.Mice with 0.03g/10g hydration after anesthesia, orbit chlorine aldehyde take blood 1.0 ml, with sterile tubes collection, centrifugal serum, blood biochemistry analyzer, the automatic detection ALT, AST level. Former says mice weight, blood collection after death mice, anatomical dislocated take liver and spleen weighing, calculation, liver and spleen visceral index. Mice executed after take liver tissue, using formaldehyde is fixed, the conventional 20% paraffin embedding slice, did HE histopathological staining light microscopy, consult immunity liver injury experimental study relevant literatures and liver tissue morphological related research literature, according to the liver tissue inflammation and necrosis and haemorrhage degree, will the liver tissue pathology change five levels.
Cut take left medial liver tissue 0.5-1.0g, cold saline flush filter paper blot, weighing, ice bath of liver homogenate made by 10%,4000 turn/min, cryogenic centrifugal 15min, fetch the supernatant fluid. According to the instruction using glucosinolate and detect malondialdehyde (mda), determining SOD activity. Its operation step by kit instructions. TNF-cactivity, IL-6 active detection by ELISA, press kit that operation. Experimental data with mean+standard deviations (X±S) said, and in groups; t test comparison:Comparison:analysis of variance between groups. P<0.05 think differences are statistically significant. With SPSS 13.0 statistical software analysis.
3.Results
(1) The treatment group ALT, AST value significantly lower than that in model group(P<0.05),indicating that NHS capsules can effectively prevent the immunity liver damage caused by the liver function decreases.
(2) The treatment group pathological structure obviously better than the model group(P<0.05), proof NHS capsules can effectively prevent the BCG add LPS of mouse liver tissue structure reexposed damage.
(3) Various treatment group liver tissue homogenate SOD was obviously higher than model group, MDA content obviously lower than model group,there is a significant difference significance(P<0.05),indicating that NHS capsule of antioxidation, scavenging free radicals role.
(4) Each treatment group TNF-α;IL-6 value obviously lower than model group,there is a significant difference significance(P<0.05), indicating that NHS capsule resistant to damage the liver function probably with its inhibition cytotoxic factor TNF-α,IL-6 the release of the relevant.
(5) Medium NHS capsule group all indexes of double ester group compared with biphenyl non-significance difference, explain various aspects and Biphenyl Dicarboxylate curative effect quite.
4.Conclusion
(1) NHS capsules can reduce BCG and LPS immunity liver damage on mice hepatic pathology structural damage and to protect its liver function.
(2) NHS capsule dose and Biphenyl Dicarboxylate curative effect quite.
引文
[1]World heath organization.Hepatitis B World Heath organization Fact Sheet 204dex.(Revised October 2000).Who Web Site http://www.who .int/mediacentre/fact sheets/fs204/en in html.
[2]徐道振.病毒性肝炎[M].北京：人民卫生出版社,2006.
[3]林永焕.乙型肝炎预防与治疗[M].上海：科学技术文献出版社,2007.
[4]Liu GT.Present status of hepatitis drugs and ets perspective In Society office of China Association for sciences and technology Research on prevention and therapy of viral hepatitis.Beijing,China SciTech Press,1991,337-341.
[5]徐叔云,卞如濂,陈修.药物试验方法学[M].北京：人民卫生出版社,2005,5：1349.
[6]冯天保.福建叶下珠抗小鼠免疫性肝损伤的实验研究[J].中药新药与临床药理,2005,16(5)：343-345.
[7]孙鸿昌.将军汤对免疫性肝损伤小鼠免疫功能影响的理论与实验研究[D].山东：山东中医药大学,2002.
[8]Wang GS,Liu GT.Role of nitric in immunologic liver damage in mice.Biochem Pharmmacol,1995,49(2):1277.
[9]杨舒淳.旋覆花苦豆子合剂应用于急性免疫性肝损伤的实验研究[D].新疆：新疆医科大学,2006.
[10]Juan Rosa,回允中.外科病理学上卷[M].北京：北京大学医学出版社,2006,9：917-943.
[11]中华医学会,传染病与寄生虫学分会,肝病学分会联合修订.病毒性肝炎防治方案[J].传染病信息,2000,13(4)：141-150.
[12]张国.抗乙肝新药拉米夫定的研究进展[J].中国新药杂志,2001, 10(2)：96.
[13]罗瑞虹.慢性乙型肝炎抗病毒治疗的现状与评价[J].国外医学内科分册,2002,29(10)：419.
[14]王颖芳.中药治疗肝损伤作用机理的实验研究进展[J].河南中医学院学报,2003,18(104)：33-34.
[15]龚非力.医学免疫学[M].北京：科学出版社,2004.
[16]颜天华.肿瘤坏死因子肝损伤机理研究进展[J].国外医学·药学分册,1998,25(6)：330-334.
[17]Tracey KJ,Cerami A.Tumor necrosis factor:a pleiotropic cytokine and therapeutic target[J].Annu Rev Med,1994,9(45):491-503.
[18]黄玲.急性肝损伤和免疫性肝损伤动物模型研究[J].广州中医药大学学报,1998,12(15)：67-69.
[19]杨天健.16种药物对卡介苗加脂多糖引起小鼠免疫性肝损伤的作用[J].中国药理学报,1997,18(2)：185-188.
[20]魏艳丽.卡介苗加脂多糖诱发小鼠免疫性肝损伤模型的研究[J].金华职业技术学院学报,2009,6(9)：3.
[21]杨舒淳.小鼠腹腔注射卡介苗加脂多糖建立急性免疫性肝损伤的实验研究[J].实验动物学,2008,25(4)：1-4.
[22]王培利.仲景和清消补法对BCG/LPS所致小鼠免疫性肝损伤免疫功能的影响[D].北京：北京中医药大学,2001.
[23]唐云安.肝损伤动物模型研究进展[J].卫生毒理学杂志,2002,16(4)：236-238.
[24]Feluga J.Laplan A.Allison AC.Protection of mice against endotoxin induced liver damage by anti-inflammatory drugs Agents Action[J]. Biochem Pharmmacol,1989,9:566.
[25]雷载权.中药学[M].上海：上海科学技术出版社,1995,1：263.
[26]葛颖华.体外培育牛黄的药理研究进展[J].中华实用中西医杂志,2006,19(19)：2322-2323.
[27]刘丽萍.熊去氧胆酸治疗肝脏疾病的作用机制和临床应用进展 [J].解放军药学学报,2004,20(4)：283-285.
[28]何秀玲.牛磺酸鹅去氧胆酸对小鼠免疫功能的影响[J].中药材,2005,28(12)：1089-1092.
[29]王天成.胆红素和牛黄拮抗正己烷致小鼠脂质过氧化作用的初步研究[J].中国工业医学杂志,2004,17(6)：356-358.
[30]蔡红娇,汪世元,刘烈刚,等.体外培育牛黄耐缺氧和清除自由基作用的研究[J].中药药理与临床药理,2003,19(6)：20-22.
[31]周建伟.牛磺酸与肝脏关系的研究进展[J].国外医学卫生学分册,1995,22(2)：97-100.
[32]曹敏.概述西洋参化学成分研究的近况[J].江苏药学与临床研究,2004,12(2)：25-26.
[33]翟鹏贵.西洋参制剂抗疲劳作用的实验研究[J].浙江中医药大学学报,2007,31(6)：761-762.
[34]王庭欣.西洋参片对小鼠抗疲劳作用的实验研究[J].食品科学,2005,26(9)：474-476.
[35]王艳宏.西洋参及其制剂的免疫调节作用研究[J].中医药学刊,2004,22(3)：566-567.
[36]赵玉珍.西洋参茎叶皂甙对大鼠实验性肝损伤的影响[J]。中成药,2003,22(3)：219-220.
[37]徐克成,孟宪铺.肝病实验室检查的临床意义[M].南京：江苏科学技术出版社,1991.
[38]邱德凯.慢性肝病临床并发症现代诊治概念[M].上海：上海科学技术出版社,2001.
[39]林绿冬.联苯双酯应用新探讨[J].黑龙江中医药,2002,5：53-54.
[40]EL-SAWY SA,el-SHAFEY AM,el-BAHRAWY HA.Effect of dime-thyl diphenyl bicarboxylate on normal and chemically-in-jured liver [J].East Mediterr Health J,2002,8(1):95-104.
[41]符铁波,刘耕陶.联苯双酯抗肝细胞损伤作用的实验研究[J].中华医学杂志,1990,7(4)：201-204.
[1]赵艳红,阮金秀。牛黄及其代用品的药理作用及临床应用[J].军事医学科学院院刊,2007,31(2)：175-177.
[2]雷载权.中药学[M].上海：上海科学技术出版社,1995.
[3]葛颖华.体外培育牛黄的药理研究进展[J].中华实用中西医杂志,2006,19(19)：2322-2323.
[4]赵文静,郝丽莉,于庆芝.实用动物药研究[M].黑龙江：黑龙江科学技术出版社,2003.
[5]李培锋,张建华,关红,等,天然牛黄、培植牛黄和鸡胆汁对大鼠足跖肿胀率及炎性组织中PEG2含量的影响[J].中草药,1998,29(9)：603-604.
[6]刘明洁,图雅,王洪涛,等.牛黄复方水煎剂的抗炎作用研究[J].中国老年学杂志,2009,29(9)：1067-1069.
[7]蔡红娇,汪世元,张喻候,等.体外培育牛黄治疗流行性乙型脑炎的临床研究[J]。华中科技大学学报,2003,32(6)：604-606.
[8]刘成德.牛黄的药理作用及临床应用概况[J].中医药信息,2006, 23(6)：14-15.
[9]王天成.胆红素和牛黄拮抗正己烷致小鼠脂质过氧化作用的初步研究[J].中国工业医学杂志,2004,17(6)：356-358.
[10]蔡红娇,汪世元,刘烈刚,等.体外培育牛黄耐缺氧和清除自由基作用的研究[J].中药药理与临床药理,2003,19(6)：20-22.
[11]汪世元,陈孝平,蔡红娇,等.体外培育牛黄诱导人肝癌HepG2细胞凋亡的实验研究[J].华中科技大学学报,2005,34(6)：754-756.
[12]仲伟玲.复方西黄胶囊治疗原发性肝癌60例[J].中国民间疗法,2003,11(9)：56-57.
[13]田彦玲,王蓓,程建新,等.牛黄天龙胶囊的抗肿瘤作用及其毒性[J].实用癌症杂志,2005,20(30)：251-253.
[14]刘丽萍.熊去氧胆酸治疗肝脏疾病的作用机制和临床应用进展[J].解放军药学学报,2004,20(4)：283-285.
[15]何秀玲,李培锋,关红.牛磺酸鹅去氧胆酸对小鼠免疫功能的影响[J].中药材,2005,28(12)：1089-1092.
[16]张能荣.胆红素[M].北京：中国医药科学出版社,1994.
[17]韦新,梁健,毛德文.牛磺酸对大鼠肝纤维化形成的影响[J]。中国中西医结合消化杂志,2004,12(1)：6-7.
[18]梁健,张锡流,杨光业.牛磺酸对大鼠肝纤维化的防治作用及机制的初步观察[J].四川大学学报：医学版,2005,36(3)：365-367.
[19]WSTERS E,WANG J H,et al.Role of taurine inpreventing acetamin-opherr induced hepatic injury in the rat[J].Am J Gastrointest Liver p-hysiol,2001,280:1274-1279.
[20]BALKAN J,PARLDAR F H,DOGRU-ABBASOGLU S,et al.The ef-fect of taurine or betaine pretreatment on hepatotoxicity and prooxi-dant status induced by lipopoly-saccharide treatment in the liver of rats[J].Eur J Gastroenterol Hepatol,2005,17(9):917-938.
[21]刘辉,金玉兰,周瑞华.牛磺酸对大鼠酒精性肝损伤保护作用的 研究[J].中国公共卫生,2003,25(3)：290-293.
[22]袁华华.牛磺酸对酒精性脂肪肝大鼠脂肪代谢的影响[J].中国兽医科技,2005,35(9)：727-730.
[231佟立权,乔海泉,周保国,等.牛磺酸对兔肝脏缺血—再灌注损伤保护作用的研究[J].中国危重病急救医学,2005,17(10)：630-631.
[24]周乾毅,袁新初,张伟.牛磺酸对肝缺血再灌注损伤过程中的糖原变化影响[J].中华综合临床医学杂志,2005,7(5)：5-6.
[25]周乾毅,袁新初,张伟.PCNA在牛磺酸对肝缺血再灌注损伤保护作用中的表达[J].中华综合临床医学杂志,2005,7(5)：13-15.
[26]DOGRU-ABBASOG LU S,KANBAGLI O,BALK AN J,et al.The p-rotective effect of taurine against thioacetamide hepatotoxicity of ra-ts[J].Human Exp Toxicol,2001,20:23-27.
[27]HAGAR H H.The protective effect of taurine against cyclosporine A induced oxidative stress and hepatotoxicity in rats[J].Toxicol Lett,2-004,151(2):335-377.
[28]梁月俭.清开灵并安宫牛黄丸治疗肺性脑病临床观察[J].中国中医急症,2006,15(6)：614-615.
[29]郭闫葵,赵世珂,孟斌.醒脑静注射液在神经内科中的应用[J].河南中医,2005,25(6)：79-81.
[30]肖兰,赵绪元.清开灵注射液的研究进展[J].中国药房,2006,17(6)：468-469.
[31]周建伟.牛磺酸与肝脏关系的研究进展[J].国外医学卫生学分册,1995,22(2)：97-100.
[2]徐道振.病毒性肝炎[M].北京：人民卫生出版社,2006.
[3]林永焕.乙型肝炎预防与治疗[M].上海：科学技术文献出版社,2007.
[4]Liu GT.Present status of hepatitis drugs and ets perspective In Society office of China Association for sciences and technology Research on prevention and therapy of viral hepatitis.Beijing,China SciTech Press,1991,337-341.
[5]徐叔云,卞如濂,陈修.药物试验方法学[M].北京：人民卫生出版社,2005,5：1349.
[6]冯天保.福建叶下珠抗小鼠免疫性肝损伤的实验研究[J].中药新药与临床药理,2005,16(5)：343-345.
[7]孙鸿昌.将军汤对免疫性肝损伤小鼠免疫功能影响的理论与实验研究[D].山东：山东中医药大学,2002.
[8]Wang GS,Liu GT.Role of nitric in immunologic liver damage in mice.Biochem Pharmmacol,1995,49(2):1277.
[9]杨舒淳.旋覆花苦豆子合剂应用于急性免疫性肝损伤的实验研究[D].新疆：新疆医科大学,2006.
[10]Juan Rosa,回允中.外科病理学上卷[M].北京：北京大学医学出版社,2006,9：917-943.
[11]中华医学会,传染病与寄生虫学分会,肝病学分会联合修订.病毒性肝炎防治方案[J].传染病信息,2000,13(4)：141-150.
[12]张国.抗乙肝新药拉米夫定的研究进展[J].中国新药杂志,2001, 10(2)：96.
[13]罗瑞虹.慢性乙型肝炎抗病毒治疗的现状与评价[J].国外医学内科分册,2002,29(10)：419.
[14]王颖芳.中药治疗肝损伤作用机理的实验研究进展[J].河南中医学院学报,2003,18(104)：33-34.
[15]龚非力.医学免疫学[M].北京：科学出版社,2004.
[16]颜天华.肿瘤坏死因子肝损伤机理研究进展[J].国外医学·药学分册,1998,25(6)：330-334.
[17]Tracey KJ,Cerami A.Tumor necrosis factor:a pleiotropic cytokine and therapeutic target[J].Annu Rev Med,1994,9(45):491-503.
[18]黄玲.急性肝损伤和免疫性肝损伤动物模型研究[J].广州中医药大学学报,1998,12(15)：67-69.
[19]杨天健.16种药物对卡介苗加脂多糖引起小鼠免疫性肝损伤的作用[J].中国药理学报,1997,18(2)：185-188.
[20]魏艳丽.卡介苗加脂多糖诱发小鼠免疫性肝损伤模型的研究[J].金华职业技术学院学报,2009,6(9)：3.
[21]杨舒淳.小鼠腹腔注射卡介苗加脂多糖建立急性免疫性肝损伤的实验研究[J].实验动物学,2008,25(4)：1-4.
[22]王培利.仲景和清消补法对BCG/LPS所致小鼠免疫性肝损伤免疫功能的影响[D].北京：北京中医药大学,2001.
[23]唐云安.肝损伤动物模型研究进展[J].卫生毒理学杂志,2002,16(4)：236-238.
[24]Feluga J.Laplan A.Allison AC.Protection of mice against endotoxin induced liver damage by anti-inflammatory drugs Agents Action[J]. Biochem Pharmmacol,1989,9:566.
[25]雷载权.中药学[M].上海：上海科学技术出版社,1995,1：263.
[26]葛颖华.体外培育牛黄的药理研究进展[J].中华实用中西医杂志,2006,19(19)：2322-2323.
[27]刘丽萍.熊去氧胆酸治疗肝脏疾病的作用机制和临床应用进展 [J].解放军药学学报,2004,20(4)：283-285.
[28]何秀玲.牛磺酸鹅去氧胆酸对小鼠免疫功能的影响[J].中药材,2005,28(12)：1089-1092.
[29]王天成.胆红素和牛黄拮抗正己烷致小鼠脂质过氧化作用的初步研究[J].中国工业医学杂志,2004,17(6)：356-358.
[30]蔡红娇,汪世元,刘烈刚,等.体外培育牛黄耐缺氧和清除自由基作用的研究[J].中药药理与临床药理,2003,19(6)：20-22.
[31]周建伟.牛磺酸与肝脏关系的研究进展[J].国外医学卫生学分册,1995,22(2)：97-100.
[32]曹敏.概述西洋参化学成分研究的近况[J].江苏药学与临床研究,2004,12(2)：25-26.
[33]翟鹏贵.西洋参制剂抗疲劳作用的实验研究[J].浙江中医药大学学报,2007,31(6)：761-762.
[34]王庭欣.西洋参片对小鼠抗疲劳作用的实验研究[J].食品科学,2005,26(9)：474-476.
[35]王艳宏.西洋参及其制剂的免疫调节作用研究[J].中医药学刊,2004,22(3)：566-567.
[36]赵玉珍.西洋参茎叶皂甙对大鼠实验性肝损伤的影响[J]。中成药,2003,22(3)：219-220.
[37]徐克成,孟宪铺.肝病实验室检查的临床意义[M].南京：江苏科学技术出版社,1991.
[38]邱德凯.慢性肝病临床并发症现代诊治概念[M].上海：上海科学技术出版社,2001.
[39]林绿冬.联苯双酯应用新探讨[J].黑龙江中医药,2002,5：53-54.
[40]EL-SAWY SA,el-SHAFEY AM,el-BAHRAWY HA.Effect of dime-thyl diphenyl bicarboxylate on normal and chemically-in-jured liver [J].East Mediterr Health J,2002,8(1):95-104.
[41]符铁波,刘耕陶.联苯双酯抗肝细胞损伤作用的实验研究[J].中华医学杂志,1990,7(4)：201-204.
[1]赵艳红,阮金秀。牛黄及其代用品的药理作用及临床应用[J].军事医学科学院院刊,2007,31(2)：175-177.
[2]雷载权.中药学[M].上海：上海科学技术出版社,1995.
[3]葛颖华.体外培育牛黄的药理研究进展[J].中华实用中西医杂志,2006,19(19)：2322-2323.
[4]赵文静,郝丽莉,于庆芝.实用动物药研究[M].黑龙江：黑龙江科学技术出版社,2003.
[5]李培锋,张建华,关红,等,天然牛黄、培植牛黄和鸡胆汁对大鼠足跖肿胀率及炎性组织中PEG2含量的影响[J].中草药,1998,29(9)：603-604.
[6]刘明洁,图雅,王洪涛,等.牛黄复方水煎剂的抗炎作用研究[J].中国老年学杂志,2009,29(9)：1067-1069.
[7]蔡红娇,汪世元,张喻候,等.体外培育牛黄治疗流行性乙型脑炎的临床研究[J]。华中科技大学学报,2003,32(6)：604-606.
[8]刘成德.牛黄的药理作用及临床应用概况[J].中医药信息,2006, 23(6)：14-15.
[9]王天成.胆红素和牛黄拮抗正己烷致小鼠脂质过氧化作用的初步研究[J].中国工业医学杂志,2004,17(6)：356-358.
[10]蔡红娇,汪世元,刘烈刚,等.体外培育牛黄耐缺氧和清除自由基作用的研究[J].中药药理与临床药理,2003,19(6)：20-22.
[11]汪世元,陈孝平,蔡红娇,等.体外培育牛黄诱导人肝癌HepG2细胞凋亡的实验研究[J].华中科技大学学报,2005,34(6)：754-756.
[12]仲伟玲.复方西黄胶囊治疗原发性肝癌60例[J].中国民间疗法,2003,11(9)：56-57.
[13]田彦玲,王蓓,程建新,等.牛黄天龙胶囊的抗肿瘤作用及其毒性[J].实用癌症杂志,2005,20(30)：251-253.
[14]刘丽萍.熊去氧胆酸治疗肝脏疾病的作用机制和临床应用进展[J].解放军药学学报,2004,20(4)：283-285.
[15]何秀玲,李培锋,关红.牛磺酸鹅去氧胆酸对小鼠免疫功能的影响[J].中药材,2005,28(12)：1089-1092.
[16]张能荣.胆红素[M].北京：中国医药科学出版社,1994.
[17]韦新,梁健,毛德文.牛磺酸对大鼠肝纤维化形成的影响[J]。中国中西医结合消化杂志,2004,12(1)：6-7.
[18]梁健,张锡流,杨光业.牛磺酸对大鼠肝纤维化的防治作用及机制的初步观察[J].四川大学学报：医学版,2005,36(3)：365-367.
[19]WSTERS E,WANG J H,et al.Role of taurine inpreventing acetamin-opherr induced hepatic injury in the rat[J].Am J Gastrointest Liver p-hysiol,2001,280:1274-1279.
[20]BALKAN J,PARLDAR F H,DOGRU-ABBASOGLU S,et al.The ef-fect of taurine or betaine pretreatment on hepatotoxicity and prooxi-dant status induced by lipopoly-saccharide treatment in the liver of rats[J].Eur J Gastroenterol Hepatol,2005,17(9):917-938.
[21]刘辉,金玉兰,周瑞华.牛磺酸对大鼠酒精性肝损伤保护作用的 研究[J].中国公共卫生,2003,25(3)：290-293.
[22]袁华华.牛磺酸对酒精性脂肪肝大鼠脂肪代谢的影响[J].中国兽医科技,2005,35(9)：727-730.
[231佟立权,乔海泉,周保国,等.牛磺酸对兔肝脏缺血—再灌注损伤保护作用的研究[J].中国危重病急救医学,2005,17(10)：630-631.
[24]周乾毅,袁新初,张伟.牛磺酸对肝缺血再灌注损伤过程中的糖原变化影响[J].中华综合临床医学杂志,2005,7(5)：5-6.
[25]周乾毅,袁新初,张伟.PCNA在牛磺酸对肝缺血再灌注损伤保护作用中的表达[J].中华综合临床医学杂志,2005,7(5)：13-15.
[26]DOGRU-ABBASOG LU S,KANBAGLI O,BALK AN J,et al.The p-rotective effect of taurine against thioacetamide hepatotoxicity of ra-ts[J].Human Exp Toxicol,2001,20:23-27.
[27]HAGAR H H.The protective effect of taurine against cyclosporine A induced oxidative stress and hepatotoxicity in rats[J].Toxicol Lett,2-004,151(2):335-377.
[28]梁月俭.清开灵并安宫牛黄丸治疗肺性脑病临床观察[J].中国中医急症,2006,15(6)：614-615.
[29]郭闫葵,赵世珂,孟斌.醒脑静注射液在神经内科中的应用[J].河南中医,2005,25(6)：79-81.
[30]肖兰,赵绪元.清开灵注射液的研究进展[J].中国药房,2006,17(6)：468-469.
[31]周建伟.牛磺酸与肝脏关系的研究进展[J].国外医学卫生学分册,1995,22(2)：97-100.
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