春柴胡质量标准化及药效学研究
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摘要
中药质量标准化研究是中药产业化、现代化、国际化的重要内容。
     柴胡是我国传统的药用植物,是临床上最常用的中药之一,具有和解表里,疏肝、升阳等功效。春柴胡为伞形科(Umbelliferae)植物狭叶柴胡Bupleurum scorzonerifolium Willd.的干燥全草。主产于江苏一带,由于春季采收,故习称“春柴胡”、“芽胡”。收载于《江苏省中药材标准》1989年版。江苏等地区习用春柴胡替代药典柴胡在临床上应用。由于春柴胡植物资源丰富,为开发利用其资源,完善其质量标准,本论文对春柴胡药材的质量标准及药效学进行了较为系统的研究。
     本论文分为三部分:第一部分文献研究;第二部分实验研究;第三部分总结与展望。
     一、文献研究:从春柴胡本草考证、柴胡属植物的研究进展及中药质量标准化现状与趋势等方面进行了文献综述,为实验研究提供思路并构筑坚实的文献基础。
     二、实验研究:主要从春柴胡的品种鉴定、化学成分、质量标准、药效学等方面进行研究,并起草了春柴胡的质量标准草案。
     1.春柴胡的品种鉴定研究:经原植物鉴定、性状鉴别和显微鉴别确定实验用药材是伞形科植物狭叶柴胡Bupleurum scorzonerifolium Willd.的春季干燥全草。
     2.春柴胡的化学成分研究:(1)用薄层色谱法对春柴胡与药典柴胡进行定性鉴别,结果春柴胡含有柴胡有效成分柴胡皂苷a、d和黄酮类成分。(2) GC-MS分析不同采收期的春柴胡挥发油化学成分发现,不同采收期的春柴胡挥发油成分接近,含量存在差异,挥发油含量以四月初至五月中旬为高。(3)不同提取方法、不同产地狭叶柴胡苗挥发油含量以CO_2超临界萃取法高于传统的水蒸气蒸馏法,挥发油化学成分的GC-MS研究表明,不同产地狭叶柴胡苗挥发油化学成分差异明显。
     3.春柴胡的质量标准研究:(1)用紫外分光光度(UV)法测定春柴胡药材中总黄酮的含量,结果药典收载的南、北柴胡在总黄酮含量上差异显著,南柴胡总黄酮含量高于北柴胡;春柴胡总黄酮含量野生高于家种;春柴胡不同采收期的总黄酮含量以四至五月为高。(2)用高效液相色谱(HPLC)法测定春柴胡药材中柴胡皂苷a、d的含量,结果药典收载的南、北柴胡在柴胡皂苷a、d含量上具有差异;春柴胡不同采收期柴胡皂苷a、d的含量差异显著,且以三至五月中旬为高。(3)通过春柴胡黄酮成分的HPLC指纹图谱(HPLC-Fingerprint Spectrum,HPLC-FPS)研究,建立了江苏句容产中药春柴胡的指纹图谱。
     4.春柴胡的药效学研究:(1)抗炎镇痛作用实验比较了春柴胡与北柴胡的挥发油抗炎镇痛作用的差异,结果春柴胡和北柴胡的挥发油均具有一定的抗炎作用,春柴胡的挥发油抗炎疗效优于北柴胡,镇痛作用二者无显著性差异。(2)通过解热作用比较春柴胡及北柴胡的解热作用,结果春柴胡及北柴胡的挥发油均具有良好的解热作用,且解热疗效春柴胡优于北柴胡。(3)春柴胡及北柴胡的水提物对急性肝损伤小鼠保护的作用实验结果显示:北柴胡5、10g/kg剂量组、春柴胡10g/kg剂量组均能显著降低肝损伤模型的ALT、AST及T-Bili。(4)春柴胡及北柴胡的水提物对二甲基亚硝胺(DMN)所致大鼠慢性肝损伤及抗肝纤维化的实验表明:春柴胡和北柴胡的水提物均具有一定的抗大鼠慢性肝损伤及抗肝纤维化的保护作用,动物的肝功能指标均有不同程度的改善,各剂量组对ALT、AST、ALP的升高和TP、ALB的降低有明显抑制作用;肝组织胶原蛋白、HA、LN含量均明显降低;改善肝脏病变程度;有良好的抗肝细胞凋亡的作用;并能下调NF-KB的mRNA的表达。北柴胡保肝疗效优于春柴胡。
     5.在春柴胡的品种鉴定、化学成分、质量标准研究结果的基础上,草拟春柴胡质量标准草案,为临床使用春柴胡提供参考。
     三、对实验结果进行分析讨论。
The standardization of quality control criteria of traditional Chinese medicine is theimportant content of industrialization,modernization and internationalization of traditionalChinese medicine.
     Bupleurum scozonerifolium Wild.(Spring Chaihu), which belongs to Umbelliferaefamily, is one of traditional Chinese medicines in common used with its entire plants.Abounding in natural resources, this species is widely distributed in jiangsu province ofChina. For it is harvested in spring, it is called"Spring Chaihu"or"Yahu"as usual,wasrecorded Chinese crude drug standard of jiangsu province(1989 edition),to develop andutilize the abundant resource, In this Paper, studies on the quality assessment andpharmacodynamics study of Bupleurum scozonerifolium Wild.
     The paper is composed of three parts.PartⅠis the literature study; partⅡisexperimental study, partⅢis summary and discussion.
     PartⅠliterature study: Review the bencaological study of Spring Chaihu, researchadvancement of Bupleurum and the standardization of quality control criteria oftraditional Chinese medicine,etc.
     PartⅡexperimental study:
     1 Variety certification study: definite the herb is the entire plants of Chun Chaihu.
     2 Chemical composition study: Spring Chaihu contains saikosaponins a,d andvolatile oils. different regions、different extraction mothed and different gathering seasonscan affect the content of volatile oils.
     3 Quality criteria study: using the UV and HPLC method, we determin the contentof saikosaponins a, d and total flavones, establishing the HPLC-Fingerprint Spectrum ofSpring Chai hu.
     4 Pharmacodynamics study: (1)Both North Chaihu and Spring Chaihu can decreaseanimal fever induced by yeast to rat and endotoxin to rabbit, but the potency of springChai hu is lower than north Chun Chaihu's, There was significant difference betweentreated groups and model groups (P<0.05). There are good antipyretic effect betweennorth Chai hu and spring Chai hu and,moreover, Spring Chai hu is better.(2)Both the high dosage in Bupleurum chinense DC and the low and high dosage in Bupleurumscorzonerifolium Willd could inhibit ear edema, reduce the abdominal capillarypermeability, whose differences in anti-inflammatory were significant compared withcontrol group (P<0.05 or P<0.01). The high dosage in both north Chahu and SpringChaihu could decrease writhing times in mice, whose differences in abirritation weresignificant compared with control group (P<0.05). The high dosage in both north Chaihu and spring Chai hu could enhance the threshold of pain. Both north Chai hu and springChai hu have showed good anti-inflammatory effects, but the latter is superior to theformer. In addition, both north Chai hu and spring Chai hu have also showed analgesiceffects, but there were no significant differences in analgesic effects between them. Theexperiment of dimethylnitrosamine (DMN)-induced chronic liver injury and anti- hepaticfibrosis in rats of noah Chai hu and Spring Chai hu show: Both north Chai hu andSpring Chai hu have protective action on chronic liver injury and anti- hepatic fibrosis inrats. Hepatic functional parameter were improved to different degrees, can significantlyinhibit the increasion of ALT、AST、ALP and loss of TP、ALB at all dose levels, thecontent of hepatic tissue collagen protein、HA、LN decreased significantly; improvedliver pathological changes; have good effect on anti-liver cell apoptosis; and downregulation the exprission of NF-KB mRNA. north Chai hu is better than Spring Chai huon protective liver action.
     5 Set up the standard of Bupleurum scozonerifolium Wild.(protocol) base on theresearch result of variety certification, chemical composition and quality criteria study.
     PartⅢAnalysis and discuss the experimental result.
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