摘要
目的:通过观察柴胡-黄芩不同比例配伍对解热抗炎作用的影响,阐明配伍对药对清热药效的影响;通过观察配伍对胆汁分泌的影响,找到最佳利胆作用的配伍比例;通过观察配伍对CCl_4所致肝损伤的保护作用,阐明配伍对药对疏肝解郁药效的影响。
方法:按照柴胡和黄芩1:0,1:2,1:1,2:1,0:1的比例制备含生药1g/ml的受试药。采用2,4-二硝基苯酚和干酵母分别制备大鼠发热模型,混合致炎液制备小鼠耳肿胀模型,分别观察五种受试药液对两种大鼠发热模型和小鼠耳肿胀模型的影响;胆总管插管引流胆汁,测定给药后3h的胆汁分泌总量,计算胆汁的体重系数。测定胆汁中总胆红素,直接胆红素,胆固醇含量,并进行比较;采用CCl4制备小鼠肝损伤模型,分别观察五种受试药液对小鼠肝损伤模型肝指数、生化指标、病理组织学、免疫组织化学等主要指标的作用。
结果:不同配伍比例受试药对2,4-二硝基苯酚致热大鼠模型的影响,与空白组比较,柴胡-黄芩1:2组和黄芩组解热作用显著,其它三组没有明显作用。对干酵母所致大鼠发热模型的影响与空白组比较,柴胡-黄芩1:1组、黄芩组、1:2组均具有明显的解热作用,2:1组和柴胡组无显著解热作用。不同配伍比例对小鼠耳肿胀度、耳肿胀率的影响:黄芩组和柴胡-黄芩1:2配伍组作用较为明显,其它三组作用与空白组比较无显著性差异;与空白组比较,黄芩组和配伍1:1组胆汁分泌量最多,其次为1:2组和2:1组,单独柴胡组无明显促进胆汁分泌的作用。与空白组比较各给药组的总胆红素水平、直接胆红素和胆固醇水平有不同程度的改变;不同配伍比例受试药对肝损伤各检测指标表现出不同的药效学作用,其中柴胡-黄芩配伍1:2组保肝作用相对更为明显,其次为黄芩组,柴胡组没有表现出明显的保护肝损伤的作用。
结论:不同配伍比例对于解热抗炎、利胆和保肝作用表现出不同的作用强度。对不同致热源所致的大鼠发热模型,其解热作用最佳配伍比例表现出一定的差异,对2,4-二硝基苯酚致热大鼠模型以1:2配伍最强,而对干酵母发热模型则以1:1配伍组表现较好。抗炎作用则主要与配伍中所含黄芩的量有关;利胆作用最佳配伍比例在柴胡-黄芩1:2和1:1之间最优;对肝损伤的保护作用,以1:2组药效学作用最为明显,其次是单独黄芩组,表明黄芩起主要保肝作用,而配伍柴胡后保肝作用更明显。
Objective:To review the anti-pyretic and anti-inflammatory effects of Radix Bupleuri and Radix Scutellare in different compatibilities,and elucidate the relationship of compatibility and pharmacodynamics;To observe the effect on bile secretion of rats administered by Radix Bupleuri and Radix Scutellare in compatibility,and look for the best proportion on Cholagogic Action;To Review the effect of Radix Bupleuri and Radix Scutellare in compatibility on hepatic injury by CCl4,and elucidate the relationship of compatibility and pharmacodynamics further.
Methods:Testing medicine of 1g/ml in accordance with the ratio 1:0,1:2,1:1,2:1,0:1 of Radix Bupleuri and Radix Scutellariae were prepared.We built rat febrile models with 2,4-dinitrophenol and dried yeast respectively and models of swelling ear in mice with mixed liquid induced inflammation.Researchers observed effects of the five testing medicine on rat febrile models and models of swelling ear in mice;Researchers detected choleresis and calculated bile-body weight after administered and formed fistula in surgery three hours,detected and compared the amount of total bilirubin,direct bilirubin and cholesterol in bile;The effects of five testing medicine on main indicators,such as liver index,biochemical indicator,histopathology and immunohistochemistry,were detected in acute hepatic injuried mouse models by CCl4 .
Results:Compared with the blank,the effect of the testing medicine in different compatibility ratio on rat febrile model by 2,4-dinitrophenol is that Radix Bupleuri:Radix Scutellariae 1:2 and 0:1 had statistical significance,and the other three groups have no obvious statistical significance.But the effect of the testing medicine in different compatibility ratio on rat febrile model by dry yeast is that Radix Bupleuri:Radix Scutellariae 1:1,0:1 and 1:2 had obvious statistical significance while the other two had no effect.Radix Bupleuri:Radix Scutellariae 0:1 and 1:2 groups had an obvious effect while the other three groups had no effect compared with the blank on tumfaction degree of mice's ears and the swelling rate;Compared with the blank,Radix Bupleuri:Radix Scutellariae 0:1 and 1:1 group had the most choleresis,1:2 and 2:1 group followed,1:0 group had no significant difference.The levels of total bilirubin,direct bilirubin and cholesterol in each drug administration group were changed in different degree;The pharmacodynamic effects of the testing medicine in different compatibility ratio was different in all investigations of hepatic injury.Radix Bupleuri:Radix Scutellariae 1:2 had the strongest hepatoprotective effect,Radix Bupleuri:Radix Scutellariae 0:1 followed while the 1:0 group had no significant conservancy effect on hepatic injury.
Conclusion:The testing medicine in different compatibility ratio has different effect size on the anti-pyretic and anti-inflammatory effects,cholagogic action and hepatoprotective effect.The best compatibility ratio is different in rat febrile models caused by two pyrogen.Radix Bupleuri:Radix Scutellariae 1:2 has the strongest effect on rat febrile model by 2,4-dinitrophenol while 1:1 on rat febrile model by dry yeast.The anti-inflammatory effects are mainly related to the amount of Radix Scutellariae in compatibility;The best compatibility ratio is between Radix Bupleuri:Radix Scutellare 1:2 and 1:1 on cholagogic action;The pharmacodynamic effects of the testing medicine in different compatibility ratio are different in all investigations of hepatic injury.Radix Bupleuri:Radix Scutellariae 1:2 has the strongest hepatoprotective effect,Radix Bupleuri:Radix Scutellariae 0:1 followed,while the 1:0 group had no significant conservancy effect on hepatic injury.That indicated Radix Scutellariae had the main hepatoprotective effect in compatibility,but the effect is stronger after compatibled with Radix Bupleuri.
引文
[1]焦树德.谈中医诊治急性病[J].河北中医,2004;26(3):165-169.
[2]高琳,白晶,刘迪谦.柴胡-黄芩水煎液中不同化学成分群配伍与其解热作用相关性研究[J].北京中医药大学学报,2006;29(11):760-764.
[3]阿荣高娃.柴胡与黄芩伍用的抗病毒作用[J].内蒙古中医药,1999;18:98.
[4]王雪峰,周丽.柴胡黄芩提取物对CVB3m感染小鼠心肌损伤影响的实验研究[J].辽宁中医杂志,2004;31(11):890-891.
[5]王宪龄,李瑞琴,吴磊.柴胡黄芩配伍不同提取部位对小鼠急性酒精性肝损伤的作用[J].中药药理与临床,2007;23(1):10-12.
[6]王宪龄,宋宁.柴胡黄芩配伍不同提取部位对大鼠急性酒精性肝损伤肝组织中环氧合酶-2表达的影响[J].中药材,2006;29(9):943-945.
[7]陈姝,梅胜尧,段晓波,等.BY1抗真菌、抗炎及镇咳祛痰作用的药效学研究[J].东南大学学报(医学版),2009;28(2):96-101.
[8]李仪奎.中药药理实验方法学[M],上海:上海科技出版社,1991,第1版:312.
[9]冯淑怡,李先荣,孙建宁.连翘酯苷抗感染、解热作用研究[J].现代生物学进展,2006;6(10):73-75.
[10]刘亚欧,白筱璐,余悦,等.柴黄制剂的解热抗炎作用研[J].中药药理与临床,2008;24(2):22-24.
[11]薛燕.柴胡的解热作用药效学研究.中医药学刊,2003;21(11):1897-1959.
[12]李仪奎.中药药理实验方法学[M],上海:上海科学技术出版社,1991,第1版:299-473.
[13]吴勃岩,高明,梁颖,等.利胆排石汤利胆作用的实验研究[J].中医药信息,2008;25(1):71-73.
[14]余成浩,文昌凡,张宏,等.清胆颗粒利胆作用的实验研究[J].四川动物,2005;24(4):627-630.
[15]朱世敏,唐志鹏.茵陈蒿汤护肝作用研究进展[J].上海中医药杂志,2008;42(2):73-75.
[16]李晓蓉,王丽娟,李宇航,等.丹芎方及其组分保肝作用的实验研究[J].中国中药杂志,2007;32(20):2177-2181.
[17] Perriseond D , Festa B.Hepatic Pharmacology:Mechanisms of Action and classification of Anfinecrotic Hepatipmtective Agents[J].Tips,1982;29:371.
[18]陈永祥,王和生,靳风云,等.一贯煎多糖对小鼠肝损伤及SOD,LPO的影响[J].中国药学杂志,1999;34(4):239-241.
[19]水祥和,舒昌杰.自由基在病毒性肝炎中的作用及其防治[J].中国实用内科杂志,1994;14(11):679-680.
[20]方允中,郑荣梁.自由基生物学的理论与应用[M],北京:科学出版社,2002,第1版:143～155.
[21]潘建秋,蒲朝煜.氧自由基与肝细胞损伤[J].贵州医药,2000;24(12):756-758.
[22]徐敏.论人体的自由基和抗氧化剂-谷胱甘肽[J].医药产业资讯,2006;3(2):127.
[23]徐春媚,王文生,曹艳红,等.连翘护肝作用的实验研究[J].黑龙江医药科学,2001;24(1):10-12.
[24]黄建康,王凤岩,黄琼,等.灵芝孢子油增强小鼠免疫功能及护肝作用的实验研究[J].现代预防医报,2007;34(7):1272-1274.
[25]林久茂,王瑞国.卷柏对实验性肝损伤小鼠保护作用的实验研究[J].福建中医学院学报,2006;16(2):28-30.
[26]陈祝英,何萍,黄仁彬,等.乙肝转阴散保护小鼠CCl4急性肝损伤的生化机理[J].中药材,2007;30(2):208-210.
[27] Macaya A. Apoptosis in the nervous system[J].Rev Neurol,1996;24(135):1356-1360.
[28] Chit tenden T,Harrington EA,Oconnor R,etal.Ind-uactio n of apop to sis by the Bcl-2 homologe Bak[J].Nature,1995;37(6524):733.
[29] Behl C,Hovey Li,Krajewsky S,etal.Bcl-2 prevents killing of neuronal cells by glutamate but not by amylord beta protein[J].Biochem Biophys Res Commun,1993;197(1):947-954.
[30] Steinman HM,Dambrine G.The bcl-2 on coprotein function as a pro-oxidant[J].Biol Chem,1995;270(4):3487-3490.
[31] Gilbert MS,Saad AH,Rupnow BA,etal.Association of bcl-2 with membranehyperpolarization and radioresistance[J].Cell Physiol,1996;168(2):114-122.
[32] Krajewski S.Investigation of subcellular distribution of bcl-2 on coprotein[J].Cancer Res,1993;53(2):4701-4702.
[33] Reed JC.Double identity for proteins of the bcl-2 family[J].Nature,1997;387:773.
[34] Apte SS,Mattei MG,Olsen BR.Mapping of the human Bax gene to chromosome and isolation of a novel alter natively spliced transcript Bax delta[J].Genom- ics,1995;26:592.
[35] Oltvai ZN,Milliman CL,Korsmeyer SJ.Bcl-2 hetero dimerizes invivo with a conserved homologue‘bax’that accelerates programmed cell death[J].Cell,1993;74:609.
[1]薛燕.柴胡的解热作用药效学研究[J].中医药学刊,2003;21(11):1897-1959.
[2]濮金龙,纪晓多,徐明,等.北柴胡挥发油的成分研究[J].化学学报,1983;41 (6):559.
[3]李廷利,都晓伟,赵景和,等.南北柴胡药理作用的比较研究[J].中国药学报,1992;12(3):234.
[4]高光,郭允珍.柴胡属12种柴胡的质量评价研究[J].中草药,1988;19(10):443.
[5]李向中.关于柴胡药理研究的探讨[J].中药通报,1983;15(2):39.
[6]周重楚,周秋丽,王本祥.北柴胡和大叶柴胡药理作用的比较[J].药学通报,1979;14(6):252.
[7]薛燕,白金叶.柴胡解热成分作用强度比较[J].中药药理与临床,2003;19(1):11.
[8] Navarro P,GinerRM,RetieM C,eta1. In vi-voAnti-inflammntory Activity of Saponins from Bupleurum rotundifolium[J]. Life Sc,2001;68(10):1199-1206.
[9]谢东浩,贾晓斌,蔡宝昌,等.北柴胡及春柴胡挥发油的抗炎镇痛作用的实验研究[J].药学与临床研究,2007;15(2):108-110.
[10]孙明瑜,谢鸣,张娜,等.小柴胡汤及其药群配伍对角叉菜胶诱导大鼠胸膜炎模型细胞因子的影响[J].中国中西医结合杂志,2004;24(7):628-631.
[11]高毅,汪爽,詹兴海.柴胡保护大鼠肝脏缺血性损伤[J].世界华人消化杂志,2000; 8(2):168-170.
[12]张征,余锋,陈先祥,等.小柴胡汤预处理对肝脏缺血再灌注损伤大鼠肝内胆管上皮细胞凋亡的影响[J].湖北中医杂志,2007;29(6):7-8.
[13]谢东浩,袁冬平,蔡宝昌,等.春柴胡及北柴胡对二甲基亚硝胺所致大鼠肝纤维化的保护作用比较[J].中国医院药学杂志,2008;28(23):2006-2009.
[14] Ushio Y,Oda Y,Abe H.Effect of saikosaponin on the im-mune responses inmice[J].Int Jmmunopharmacol,1991;13(5):501.
[15]梁云,崔若兰.柴胡皂甙及其同系物抗炎和免疫功能的研究进展[J].中国中西医结合杂志,1998;18(7):446.
[16] Kato M,Pu MY,Isobe KI,etal. Characterization of the immuno regulatory action of Saikosaponind[J]. Cell Immunol,1994;159(1):15.
[17] Motoo Y,Sawabu N.Antitumor effects of saikosaponins,baicalin and baicalein on human hepatoma cell lines[J].Cancer Lett,1994;86(1):91.
[18] Chiang LC,Ng LT,Liu LT,etal. Cytotoxicity and anti-hepatitis B virus activities of saikosaponins from Bupleurum species[J]. Planta Med,2003;69(8):705.
[19] Tsai YJ,Chen IL,Horng LY,etal. Induction of differention in rat C6 glioma cells with Saikosaponins[J].Phytother Res,2002;16(2):117.
[20]步世忠,许金康,孙继虎,等.柴胡皂甙d上调人急性早幼粒白血病细胞糖皮质激素受体mRNA对细胞生长的影响[J].中国中西医结合杂志,2000;20(5):350.
[21] Ahn BZ,Yoon YD,Lee YH,etal. Inhibitory effect of bupleuri radix saponins on adhesion of some solid tumor cells and re-lation to hemolytic action: screening of 232 herbal drugs for anti-cell adhesion[J]. Planta Med,1998;64(3):220.
[22]夏薇,崔新羽,崔清谭.柴胡皂甙d(SSd)对K562细胞增殖的抑制作用[J].北华大学学报,2002;3(2):113.
[23]王丽娟,王勇,朱旭燕,等.黄芩抗炎镇痛作用的实验研究,齐齐哈尔医学院学报,2008;29(11):1304-1306.
[24]王浴生,邓文龙,薛春生.中药药理与临床[M],北京:人民卫生出版社,1998,第2版:972-982
[25]徐叔云,丁长海,魏伟,等.影响炎症免疫反应的药物研究及其思路[J].药学进展,1997;15(2):77.
[26]戴岳,张聪,林已笼,等.黄芩总苷对大鼠急慢性前列腺炎影响的实验研究[J].中医药学刊,2003;21(3):386-387.
[27]郑红,周新灵,明彩荣,等.黄芩苷、枳壳抗肥大细胞脱颗粒的实验研究[J].中国中医基础医学杂志,2005;11(6):434.
[28]顾红缨,郭焱,罗晶.黄芩甙对I型变态反应豚鼠超微结构的影响[J].长春中医学院学报,2002;18(4):40-41.
[29]刘云波,郭丽华,邱世翠,等.黄芩体外抑菌作用研究[J].时珍国医国药,2002;13(10):5961.
[30]邝枣园,黄衍寿,吴伟,等.黄芩苷对肺炎衣原体诱导的内皮细胞粘附因子表达的影响[J].广州中医药大学学报,2004;21(6):4541.
[31]邝枣园,黄衍寿,吴伟.黄芩苷对肺炎衣原体诱导的可溶性细胞粘附因子及IL-8的影响[J].浙江中医杂志,2004;39(11):5021.
[32]赵宝珍,李仲兴,王秀华,等.五倍子等5种中药水煎剂对表皮葡萄球菌的抗菌活性的观察[J].中国感染控制杂志,2004;3(3):211-213.
[33]韩喻美.金丝桃甙、黄芩甙对小鼠红细胞超氧化物岐化酶的影响[J].中国中医药科技,2002;9(5):2841.
[34]刘玉萍,Purusotam Basnet,曹晖,等.黄芩清除自由基活性与黄芩苷含量的相关性研究[J].中国中药杂志,2002;27(8):575-5791.
[35]王汉渝,洗超贵,钟睿,等.黄芩中酚性苷类对腮腺放射损伤的防护作用的研究[J].肿瘤防治杂志,2003;10(6):637-639.
[36]骆丹,闵玮,林向飞,等.羟氯喹及黄芩等中药对紫外线损伤角质形成细胞的保护作用[J].中国美容医学,2003;12(4):355-358.
[37]汪晓军,马贳,张奉学,等.黄芩苷对ConA致肝损伤小鼠肝组织MDA含量的影响[J].世界华人消化杂志,2005;13(9):1153-1155.
[38]陈桂枝,罗德生,郑红花,等.四氯化碳肝损伤与脂质过氧化时黄芩煎剂保护作用[J].成宁学院学报(医学版),2005;19(1):331-331.
[39]卢春凤,王丽敏,陈廷玉.黄芩素和黄芩苷对四氯化碳所致肝脏损伤大鼠转氨酶的影响[J].黑龙江医药科学,2003;26(4):50-511.
[40]李素婷,杨鹤梅,石艳华,等.黄芩茎叶总黄酮保肝作用实验研究[J].中国中医药信息杂志,2004;11(7):593-5941.
[41]李素婷,梅立新,杨鹤梅,等.黄芩茎叶总黄酮对实验性大鼠肝纤维化的影响[J].辽宁中医杂志,2004;31(10):871-8721.
[42]林小聪,刘新光,陈伟珠.黄芩素和大豆黄酮对人白血病细胞系HL-60体外增殖的抑制作用[J].右江医学,2004;3(2):93-95.
[43]罗德生,郑红花,刘琴,等.黄芩煎剂对大鼠CCl4急性肝损伤保护作用的实验研究〔J〕.咸宁医学院学报,2001;15(2):92-94.
[44]李素婷,石艳华,杨鹤梅,等.黄芩茎叶总黄酮对肝脏MDA生成的影响[J].中国中医药信息杂志,2001;8(6):23-24.
[45]曾雅静,陈韵.黄芩素对人乳腺癌MCF-7细胞内酪氨酸蛋白激酶的抑制作用[J].医学文选,2005;24(4):473-475.
[46]尹飞,杨于嘉,虞佩兰,等.黄芩甙对百日咳菌液致离体大鼠脑组织损害的保护作用及量效关系的研究[J].中国中西医结合杂志,2002;22(4):286-288.
[47]陈忻,董晓丹,张楠,等.黄芩苷对局灶性脑缺血模型大鼠的保护作用[J].广州中医药大学学报,2006;23(6):506-508.
[48] Shinichi Ikemoto,Kaiunobu Sugimuka,Naomasa Yo shida,etal.Anxiolytie-like effect s of baicalein and baicalin in t he vogel conflict testin mice[J]. Eur J Pharmacal,2003;464(2):141-146.
[49]焦树德.谈中医诊治急性病[J].河北中医,2004;26(3):165-169.
[50]高琳.柴胡-黄芩不同化学成分群配伍与退热保肝药效相关性研究[J].北京中医药大学硕士学位论文,2003.
[51]阿荣高娃.柴胡与黄芩伍用的抗病毒作用[J].内蒙古中医药,1999;18(增刊):98-99.
[52]王雪峰,周丽.柴胡-黄芩提取物对CVB3m感染小鼠心肌损伤影响的实验研究[J].辽宁中医杂志,2004;31(11):890-891.
[53]王宪龄,穆海博,刘仁慧,等.柴胡黄芩不同配伍比例干预急性酒精性肝损伤大鼠肝脾、胸腺、肾上腺指数及肾上腺中维生素C含量的变化[J].中国临床康复,2006;10(15):182.
[54]王宪龄,程玲,宋宁,等.柴胡黄芩及两者不同比例配伍抗大鼠CCl4慢性肝损伤作用的比较研究[J].中药药理与临床,2009;25(6):70-73.