藿胆丸治疗鼻炎的药效物质基础研究
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摘要
中药药效物质基础研究是开发有效成分明确、疗效确切、质量稳定可控的新一代中成药基础,是中药实现现代化的关键。中药物质基础研究的方法有很多,本课题采用分离重组思路对藿胆丸物质基础进行研究。根据藿胆丸组方中广藿香叶和猪胆粉化学组成、组方特点,合理采用现代工业技术,获取广藿香叶和猪胆粉中多种化学组分,以传统丸剂为参照,进行抗炎抗过敏化学组分分析跟踪,探明藿胆丸抗炎抗过敏药性成分为广藿香油和猪去氧胆酸;以广藿香油和猪去氧胆酸为组方成分,进行配伍和剂量的优化研究,为研发新一代中药新药奠定前期研究基础。
藿胆丸由广藿香叶和猪胆粉两味组成。藿胆丸源于清代《医宗金鉴》,是治疗慢性鼻炎、慢性鼻窦炎的名优中成药,具有芳香化浊,清热通窍的功效,临床主要用于治疗湿浊内蕴、胆经郁火所致的鼻塞、流清涕、前额头痛等症,已有二百余年的临床应用历史。藿胆丸疗效确切,但产品粗糙,存在生产工艺陈旧、剂型落后、服用量大等不足。本课题开展了如下研究:
1、藿胆丸中广藿香叶有效部位筛选
以抗炎和抗过敏药效试验筛选广藿香叶有效部位,并以成药藿胆丸为对照,结果表明,广藿香叶水溶性部位和不含挥发油的脂溶性部位虽有抗炎作用,但作用强度明显低于挥发油部位、广藿香叶、藿胆丸成药;广藿香叶水溶性部位和不含挥发油的脂溶性部位不具有抗过敏作用,而挥发油部位与广藿香叶、藿胆丸具有抗过敏作用。广藿香叶挥发油同时具有抗炎、抗过敏作用,其作用强度接近广藿香叶、藿胆丸成药,因此可确定藿胆丸中广藿香叶有效部位主要是广藿香挥发油。
比较超临界二氧化碳萃取(以下用SFE-CO_2表示)和水蒸汽蒸馏的方法提取广藿香叶挥发油。结果表明,采用水蒸汽蒸馏,只能得芳香水,甚难得到挥发油。故本课题采用SFE-CO_2萃取技术提取广藿香叶挥发油并确定其提取工艺:广藿香叶粉碎成粗粉,SFE-CO_2提取,提取参数为萃取温度53℃,萃取压力13MPa,解析压力5.5MPa,解析温度63℃,二氧化碳(CO_2)流量为400L/h,萃取时间为2.0小时,得广藿香叶挥发油。
采用SFE-CO_2萃取所得广藿香叶挥发油,与市售广藿香油比较,两者质量一致,均符合2005年版中国药典规定。
2、广藿香油对藿胆丸药效学的影响
用广藿香油替代藿胆丸中的广藿香叶药材所得的样品称为精制藿胆方,即精制藿胆方的组成为广藿香油和猪胆粉。比较精制藿胆方、藿胆丸、广藿香油、猪胆粉的药效学,结果表明:
(1)精制藿胆方、广藿香油、猪胆粉与藿胆丸均具有抗炎作用。抗炎指标有醋酸所致小鼠腹腔毛细血管通透性增高、角叉菜胶所致大鼠足趾肿胀、二甲苯所致小鼠耳廓肿胀,大鼠棉球肉芽肿等四种急性和亚急性炎症模型。精制藿胆方对大鼠角叉菜胶所致足趾肿胀、二甲苯所致小鼠耳廓肿胀等急性炎症模型、大鼠棉球肉芽肿的亚急性炎症模型的抑制作用强度与藿胆丸相当,对抑制醋酸所致小鼠毛细血管通透性增高作用优于藿胆丸。广藿香油、猪胆粉两者配伍后协同增效作用明显。提示用广藿香油替代藿胆丸中的广藿香叶,可以保持藿胆方的抗炎作用。
(2)精制藿胆方、广藿香油、猪胆粉与藿胆丸原方均具有抗过敏作用。抗过敏指标有卵蛋白致敏豚鼠离体回肠肌的过敏性收缩反应,同种大鼠被动皮肤过敏反应(PCA),2,4—二硝基氯苯所致小鼠耳廓皮肤迟发型超敏反应等过敏反应模型。精制藿胆方对卵蛋白致敏豚鼠离体回肠肌的过敏性收缩反应、同种大鼠被动皮肤过敏反应(PCA)的抑制作用优于藿胆丸;对2,4—二硝基氯苯所致小鼠耳廓皮肤迟发型超敏反应的抑制作用强度相当于藿胆丸。广藿香油、猪胆粉两者配伍后协同增效作用明显。提示用广藿香油替代藿胆丸中的广藿香叶,可以保持藿胆方的抗过敏作用。
(3)精制藿胆方具有体内外抗菌作用。精制藿胆方体外抗菌试验对所试菌种(金黄色葡萄球菌、甲型溶血性链球菌、乙型溶血性链球菌、肺炎链球菌、卡他球菌、白喉棒状杆菌、大肠埃希氏菌、绿脓假单胞菌及白色念珠菌)均有不同程度的抗菌作用;对感染肺炎双球菌、乙型溶血性链球菌、金黄色葡萄球菌的小鼠,均具有显著的降低死亡率的作用。提示用广藿香油替代藿胆丸中的广藿香叶,可以保持藿胆方的体内外抗菌作用。
(4)精制藿胆方、广藿香油、猪胆粉与藿胆丸均具有增强免疫作用。增强免疫作用指标有小鼠网状内皮系统碳粒廓清吞噬作用的模型。精制藿胆方、广藿香叶挥发油、猪胆粉均对小鼠网状内皮系统碳粒廓清的吞噬作用具有显著增强作用;精制藿胆方增强免疫作用强度与藿胆丸相当。提示用广藿香油替代藿胆丸中的广藿香叶,可以保持藿胆方的增强免疫作用。
研究结果表明,由广藿香油和猪胆粉组成的精制藿胆方具有抗炎、抗过敏、抗菌、增强免疫作用;广藿香油与猪胆粉两者配伍应用,在抗炎、抗过敏药效中,具有显著的协同增效作用;在抗炎、抗过敏、抗菌、增强免疫各药效指标中,精制藿胆方相当于甚至优于藿胆丸。结果提示,藿胆丸中的广藿香叶以广藿香油替代,可保持原藿胆丸的药效。
3、猪胆粉化学成分对肝功和血液生化指标的影响
由广藿香油和猪胆粉组成的精制藿胆方中,仍以猪胆粉直接入药,从而造成成型困难、气味难闻、卫生学难过关、质量难监控,同时由于猪胆粉含一定量的胆固醇,长期服用可能会引起血中胆固醇偏高等缺陷,迫切需要利用现代科学技术寻找出与其药效等同的有效部位或有效成分取代。
猪去氧胆酸是猪胆粉的主要有效成分,具有化学成分单一,质量稳定可控,同时不含胆固醇、杂质少、易成型等优势。现代药理研究发现,胆酸类成分是猪胆粉多方面药理作用的有效成分,尤其是在抗炎、抗过敏、抗菌及增强免疫方面,胆酸类成分的作用表现更为突出。而猪胆粉中的胆酸类成分主要含猪去氧胆酸,为猪去氧胆酸替代藿胆丸中的猪胆粉提供了理论依据。
用广藿香油和猪去氧胆酸组成藿胆有效成方,即藿胆有效成方的组成为广藿香油和猪去氧胆酸。比较精制藿胆方和藿胆有效成方对血中胆固醇及肝功能损害的影响。结果表明,药后90天,精制藿胆方的血中胆固醇、ALP、AST水平均高于空白对照组;精制藿胆方ALT的水平与空白组相当;停药14天,精制藿胆方血中胆固醇、ALP、AST、ALT水平均恢复正常;藿胆有效成方给药后90天血及停药14天血中胆固醇、ALP、AST、ALT水平均与空白对照组相当。实验结果提示,精制藿胆方长期用药会导致血中胆固醇及部分转氨酶偏高,藿胆有效成方则可避免该结果。猪去氧胆酸替代猪胆粉应用于藿胆丸中,可以避免对血中胆固醇及肝功能损害的影响,使其安全性得到更好的保障。
4、猪去氧胆酸对藿胆丸药效学的影响
藿胆有效成方能够避免对血中胆固醇及肝功能损害的影响,是否能保持藿胆丸的药效尚需实验证实。以抗炎、抗过敏、抗菌及免疫功能试验等指标,来评价猪去氧胆酸的替代是否可行。结果表明:
(1)藿胆有效组分方、猪去氧胆酸、广藿香油、精制藿胆方、猪胆粉及藿胆丸原方均对醋酸所致小鼠腹腔毛细血管通透性增高、二甲苯所致小鼠耳廓肿胀,角叉菜胶所致大鼠足趾肿胀、大鼠棉球肉芽肿均有明显抑制作用。藿胆有效组分方对小鼠腹腔毛细血管通透性增高、二甲苯所致小鼠耳廓肿胀急性炎症模型的抑制作用与藿胆丸原方、精制藿胆方相当。藿胆有效组分方对角叉菜胶所致大鼠足趾肿胀、大鼠棉球肉芽肿的抑制作用略优于藿胆丸原方及精制藿胆方。猪去氧胆酸对醋酸所致小鼠腹腔毛细血管通透性增高、二甲苯所致小鼠耳廓肿胀,角叉菜胶致大鼠足趾肿胀、大鼠棉球肉芽肿的抑制作用与猪胆粉相当。猪去氧胆酸、广藿香油单独应用均有较好的抗炎作用,且两药配伍有协同增效的抗炎作用。实验结果提示,用猪去氧胆酸替代藿胆丸中的猪胆粉、可以保持藿胆方的抗炎作用。
(2)藿胆有效组分方、猪去氧胆酸、广藿香油、精制藿胆方、猪胆粉及藿胆丸原方均对卵蛋白致敏豚鼠离体回肠肌的过敏性收缩反应,具有快速抑制作用;对同种大鼠被动皮肤过敏反应(PCA)具有明显的抑制作用;对2,4—二硝基氯苯所致小鼠耳廓皮肤迟发型超敏反应有明显的抑制作用。藿胆有效组分方对卵蛋白致敏豚鼠离体回肠肌的过敏性收缩反应,对同种大鼠被动皮肤过敏反应(PCA),对2,4—二硝基氯苯所致小鼠耳廓皮肤迟发型超敏反应的抑制作用强度与精制藿胆方、藿胆丸原方相当。猪去氧胆酸对卵蛋白致敏豚鼠离体回肠肌的过敏性收缩反应,对同种大鼠被动皮肤过敏反应(PCA),对2,4—二硝基氯苯所致小鼠耳廓皮肤迟发型超敏反应的抑制作用强度与猪胆粉相当。猪去氧胆酸、广藿香油单独应用均有较好的抗过敏作用,且两药配伍有协同增效的抗过敏作用。实验结果提示,用猪去氧胆酸替代藿胆丸中的猪胆粉、可以保持藿胆方的抗过敏作用。
(3)藿胆有效组分方、猪去氧胆酸、广藿香油、精制藿胆方、猪胆粉及藿胆丸原方均能显著增强小鼠网状内皮系统碳粒廓清的吞噬作用,藿胆有效组分方对小鼠网状内皮系统碳粒廓清的吞噬功能的作用强度与藿胆丸原方、精制藿胆方相当。猪去氧胆酸对小鼠网状内皮系统碳粒廓清的吞噬功能的作用强度与猪胆粉相当。猪去氧胆酸、广藿香油单独应用均有较好的增强免疫功能的作用,且两药配伍有协同增效的增强免疫功能的作用。实验结果提示,用猪去氧胆酸替代藿胆丸中的猪胆粉、可以保持藿胆方的增强免疫功能的作用。
(4)藿胆有效组分方、精制藿胆方体外抗菌试验对所试菌种(金黄色葡萄球菌、甲型溶血性链球菌、乙型溶血性链球菌、肺炎链球菌、卡他球菌、白喉棒状杆菌、大肠埃希氏菌、绿脓假单胞菌及白色念珠菌)的MIC,藿胆有效组分方介于9.8×10-5g/mL～2.0×10-1g/mL之间,精制藿胆方介于6.2×10-3g/mL～2.0×10-1g/mL之间,提示藿胆有效组分方体外抗菌作用强度比精制藿胆方强。藿胆有效组分方、精制藿胆方对感染肺炎双球菌混悬液、乙型溶血性链球菌混悬液、金黄色葡萄球菌混悬液的小鼠的死亡率有不同程度的抑制作用,藿胆有效组分方对感染肺炎双球菌混悬液、乙型溶血性链球菌混悬液、金黄色葡萄球菌混悬液的小鼠的死亡率的抑制作用明显优于精制藿胆方。提示猪去氧胆酸替代藿胆丸中的猪胆粉,更有利于其抗菌作用的发挥。
通过对藿胆丸拆方研究广藿香叶和猪胆粉的有效部位或有效成分,并与藿胆丸原方进行比较,明确了藿胆丸原方产生临床疗效的有效部位是由广藿香油和猪去氧胆酸,且二者之间具有协同增效的作用。采用藿胆丸有效成分组方不但可以避免藿胆丸原方的不良反应,而且从根本上克服传统藿胆丸剂量大、成型难,成品质量波幅大等技术问题,提高了藿胆丸的技术含量,为产品质量达到“安全、有效、稳定、可控”提供技术支撑。
本研究明确了藿胆丸治疗鼻炎的物质基础,用广藿香油和猪去氧胆酸组成的处方,具有药效作用物质明确,能够达到现代化中成药的雏形,为藿胆丸的产品升级换代奠定理论基础,提供了科学的依据。本研究方法可作为阐明中药复方物质基础提供一种科学的研究思路和方法。
Studying on the substance foundation of Chinese Herb Pharmacodynamic is the basis of research and development a new generation Chinese Patent Medicine with definite effective components,exact efficacy,stable and controllable quality,also the key to achieve modernization of Chinese traditional medicine.To investigate the substance foundation of Huodan pill by means of the Purification & Recombinant is the innovation in this project. According to the chemical composition and the feature in this formula,we could apply modern industrial technology to get various chemical compositions of Herba Pogostemonis' leaf and Porcine Gall Powder,and verify the active components by tracing and analysis the anti-inflammatory and anti-allergic chemical compositions with reference to traditional pill, which were used as compounds to Investigation on compatibility and dose optimization.It can lay a foundation of previous research for developing a new traditional Chinese medicine.
Huodan pill,rooted from Yi Zong Jin Jian in Qing dynasty,is a famous Chinese Patent Medicine treating chronic rhinitis and chronic sinusitis.With the efficacy of resolving turbidity with aroma,clearing heat and relieving stuffiness,it can be used to treating intrinsic damp turbidity,nasal obstruction,nasal discharge and prefrontal ache accused by stagnant fire in Dan channel.It has a clinical application history more than two hundred years.Huodan pill is consist of two parts:Herba Pogostemonis' leaf and Porcine Gall Powder.Although the definite therapeutic effect,there are a large of disadvantages such as the rough product,old production process,backward dosage forms.The project developed a series of research as below:
1 Screening of active part ofHerba Pogostemonis' leaf in Huodan pill. We screened active part of Herba Pogostemonis' leaf by pharmacodynamic test used in anti-inflammation and anti-hypersensitiveness drug,and used patent medicine Huodan pill as the control group.The results demonstrated that hydrophilic and lipophilic part without volatile oil of Herba Pogostemonis' leaf have the effect of anti-inflammation,but lower than volatile oil,total extraction of Herba Pogostemonis' leaf and the control group.At the same time,both parts have little effect of anti-hypersensitiveness.While volatile oil of Herba Pogostemonis' leaf have the same anti-inflammation effect as total extraction of Herba Pogostemonis' leaf and the control group,as well as the anti-hypersensitiveness effect.We concluded that volatile oil is the main active part of Herba Pogostemonis' leaf in Huodan pill.
We extracted volatile oil from Herba Pogostemonis' leaf by two methods:SFE-CO2 and distillation,and the results showed that only aromatic water can be gotten by the method of distillation.So the SFE-CO_2 is adopted to extract volatile oil.The protocol of extraction is the following:Herba Pogostemonis' leaf is crashed to coarse powder first,and then extracted by SFE-CO_2.The extract temperature is 53℃,extract pressure is 13MPa,while the resolve pressure is 5.5Mpa,the resolve temperature is 63℃,the flow rate of CO_2 is 400L/h.The whole process lasted 2 hours.And then the volatile oil can be gotten. Volatile oil extracted by SFE-CO_2 accord to Herba Pogostemonis' oil in market,and both are consistent with Chinese Pharmacopeia.
2 Evaluation ofHerba Pogostemonis' oil in Huodan pill. Refined HuoDan Recipe is comprised ofHerba Pogostemonis' oil and Porcine Gall Powder. We studied pharmacological effect of the refined HuoDan Recipe,Huodan pill,Herba Pogostemonis' oil,Porcine Gall Powder and results as showed below.
a) Refined Huodan Recipe,Herba Pogostemonis' oil,Porcine Gall Powder and Huodan pill all have anti-inflammatory effects.The indexes include increasing capillary permeability of mouse belly cavity,swelling of rat toes by carrageenan, swelling of mouse auricle by dimethl benzene and rat cotton ball granuloma.,the acute and subacute inflammation model.Refined Huodan Recipe has the same inhibitory effect on the subacute inflammatory model such as swelling of rat toes by carrageenan,swelling of mouse auricle by dimethl benzene and rat cotton ball granuloma as Huodan pill,and better inhibitory effect on increasing capillary permeability of mouse belly cavity by acetic acid than Huodan pill.Herba Pogostemonis' oil and Porcine Gall Powder,synergize the anti-inflammatory effect, which implies that refined Huodan recipe has good anti-inflammatory effect as well as compositive materials,and which have synergism.
b) There are all anti-allergy effect in Refined Huodan Recipe,Herba Pogostemonis'oil,Porcine Gall Powder and original prescription of Huodan pill. The indexes include allergia contractile response of ileum muscle in cobaya by egg albumin,passive cutaneous ana-phylaxis reaction(PCA),delayed hypersensitivity of mouse's pinna skin by 2,4-dinitro-chlorobenzene.Refined Huodan Recipe has higher depressant effect on Allergic Contractile response of cony pig by Ovalbumin and PCA of same species of rat than Huodan pill.The results have indicated that Refined Huodan Recipe and its disassembled prescriptions have remarkable anti-allergy effects.Drug synergy has the relationship of synergistic action.
c) Refined Huodan Recipe has significant anti-microbial doughty effects.Refined Huodan Recipe has obvious dispatity degree anti-bacterial action against staphylococcus aureus,alpha hemolytic streptococcus,beta-hemolytic streptococci, streptococcus pneumoniae,micrococcus catarrhalis,corynebacterium diphtheriae, escherichia,aeruginosus bacillus,blastomyces albicans;The in corpore anti-microbial tests showed that Refined Huodan recipe has doughty inhibit activity to diplococcus lanceolatus,staphylococcus aureus,beta-hem olytic streptococci.These research results indicate that Refined Huodan recipe has anti-microbial doughty effects.
d) Refined Huodan Recipe,Herba Pogostemonis'oil,Porcine Gall Powder and Huodan pill have significant enhancing immunity function effects.Drug synergy has the relationship of synergistic action.Refined Huodan Recipe,Herba Pogostemonis'oil,Porcine Gall Powder can obvious strengthen the phagocytosis of mice's reticuloendothelial system.Refined Huodan Recipe's enhancing immunity function effect is as the same as Huodan pill.The results have indicated that Refined Huodan Recipe and its disassembled prescriptions have remarkable enhancing immunity function effects.
The results of the experiment indicated that Refined Huodan Recipe,which made of Herba Pogostemonis'oil and Porcine Gall Powder,havs notable anti-inflammatory effects, anti-allergy effects,anti-microbial effects and enhancing immunity function effects.Drug synergy has the relationship of synergistic action.The results show that substitute Herba Pogostemonis'oil for Herba Pogostemonis' leaf,Refined Huodan Recipe can reserve its effect.
3 Effect of Porcine Gall Powder composition for liver function and blood biochemical. Because of Porcine Gall Powder,the Huodan Pills have the disadvantage of difficult forming,the older preparation,large taken dose,foul smells and difficult controlling of the microorganism.In addition,Because of the amount of cholesterol and Chenodeoxycholic acid in Porcine Gall Powder,you will have cell toxicity,abnormal determination of cholesterol in your blood and liver function damage,if you take in a long time of Huodan pills.In order to explore a kind of preparation that has the advantage of defined effective medium,stable quality,well-defined Pharmacological effect,well-defined therapy and normal determination of cholesterol in your blood,although you take in a long time of them. It is very necessary to look for the active site or the effective component of the Porcine Gall Powder,which can Substitute of Porcine Gall Powder Using in Huodan Pills.The Cholic acid has many kinds of Pharmacological effect,especially in anti-inflammatory,anti-allergy, anti-microbial and enhancing immunity function effects.The other Cholic acid is very little, no notion of Pharmacological effect and some are hurt to liver function and cardiovascular system.
Hyodeoxycholic acid is the main active component of Porcine Gall Powder,which has testified by modern research.It has the advantage of one chemical compound,stable quality, clearing Pharmacological effect,no foreign matter and no the amount of cholesterol and easy forming,which offers the theoretical support to Hyodeoxycholic acid substitute of Porcine Gall Powder using in Houdan pills.
Porcine Gall Powder recipe can increase the level of cholesterol in blood and reduce the damage of the liver function,but Hyodeoxycholic acid recipe can not,if you take in a long time of them.The levels of cholesterol,AST、ATP in blood of the Porcine Gall Powder recipe low,middle,high dose groups is higher than the blank group,the level of ALT as the same as the blank group,after giving drug for 90 days.The levels of cholesterol,AST,ATP in blood of the Porcine Gall Powder recipe low,middle,high dose groups come back into normal level after stopping using drug for 14 days.The levels of cholesterol,AST、ATP、ALT in blood of the Hyodeoxycholic acid recipe low,middle,high dose group is the same as the blank group,after giving drug for 90 days and stopping using drug for 14 days. The results of the experiment indicate that using Hyodeoxycholic acid Substituting of Porcine Gall Powder Using in Huodan Pills makes to raise the safety of Huodan Pills.
4 Effect of Hyodeoxycholic acid for the pharmacodynamics of Huodan pill This effective compound recipe consists of Herba Pogostemonis'oil and Hyodeoxycholic acid could avoid from injuring the liver,but it can be still effective as Huodan pill? Four kinds evaluating indicator of rhinitis such as anti-inflammatory,anti-allergy,anti-bacterial and enhancing immunity function experiment were used to compare the pharmacology of the Hyodeoxycholic acid instuting ofPulvis Fellis Suis before and after.The conclusion is:
a) Huodan effective component recipe,Hyodeoxycholic acid,Herba Pogostemonis'oil, Refined Huodan Recipe,Porcine Gall Powder and Huodan pill has significant anti-inflammatory effects.Hyodeoxycholic acid and Oleum pogostemonis have the relationship of synergistic action.Hyodeoxycholic acid recipe has the same inhibitory action on mice's auricle swelling induced by xylene,the increase of capillary permeability in mice induced by acetic acid,rat's toes swelling induced by carrageenan, the granuloma by the cotton ball as Huodan pills and Porcine Gall Powder recipe. Hyodeoxycholic acid has the same inhibitory action on mice's auricle swelling induced by xylene,the increase of capillary permeability in mice induced by acetic acid,rat's toes swelling induced by carrageenan,the granuloma by the cotton ball as Porcine Gall Powder.Hyodeoxycholic acid and Oleum pogostemonis have significant anti-inflammatory effects.Drug synergy has the relationship of synergistic action.The results of the experiment indicate that Hyodeoxycholic acid is the active component of the Porcine Gall Powder in anti-inflammatory pharmacological effects and Hyodeoxycholic acid may be institute of Porcine Gall Powder using in Huodan Pills.
b) Huodan effective component recipe,Hyodeoxycholic acid,Herba Pogostemonis'oil, Refined Huodan Recipe,Porcine Gall Powder and Huodan pill has the relationship of synergistic action.Hyodeoxycholic acid,Oleum pogostemonis and Huodan recipe have obvious inhibitory action on extracorporeal ileum muscles' hypersensitive contractility reaction of Guinea pig sensitized by albumin,homogeneous passive cutaneous anaphylaxis in mice,mice's auricle cutaneous forme tardive supersensitivity action by 2,4-dinitrochlorobenzene.The results have indicated that Huodan recipe and its disassembled prescriptions have remarkable anti-allergy effects.Drug synergy has the relationship of synergistic action.
c) Huodan effective component recipe,Hyodeoxycholic acid,Herba Pogostemonis'oil, Refined Huodan Recipe,Porcine Gall Powder and Huodan pill have significant enhancing immunity function effects.Drug synergy has the relationship of synergistic action.Hyodeoxycholic acid,Oleum pogostemonis and Huodan recipe can obvious strengthen the phagocytosis of mice's reticuloendothelial system.The results have indicated that Huodan recipe and its disassembled prescriptions have remarkable enhancing immunity function effects.Drug synergy has the relationship of synergistic action.
d) Huodan effective component recipe,Hyodeoxycholic acid,Herba Pogostemonis'oil, Refined Huodan Recipe,Porcine Gall Powder and Huodan pill has significant anti-microbial doughty effects.Huodan recipe has obvious dispatity degree anti-bacterial action against staphylococcus aureus,alpha hemolytic streptococcus, beta-hemolytic streptococci,streptococcus pneumoniae,micrococcus catarrhalis, corynebacterium diphtheriae,escherichia,aeruginosus bacillus,blastomyces albicans; The in corpore anti-microbial tests showed that refined huodan recipe has doughty inhibit activity to diplococcus lanceolatus,staphylococcus aureus,beta-hem olytic streptococci.Huodan recipe has anti-microbial doughty effects.
By studying on the effective fractions or components of Herba Pogostemonis' leaf and Porcine Gall Powder respectively,and comparing with the original formula,it could affirm that the effective parts are patchouli oil and hyodeoxycholic acid which have the synergistic interaction.Formulating with the effective compositions of Huodan pill not only could avoid the adverse reaction of the original prescription,but also overcoe the technical problems fundamentally such as large dosage,hard shaping and unstable quality.It offered a technological support for the high quality product of "safe,effective,stable,and controllable".
This project has made clear the material basis of treating rhinitis.The formula consists of patchouli oil and hyodeoxycholic acid is a rudiment of the modern Chinese patent medicine. It provided scientific basis and laid a theoretical foundation for product renewal.This study method could offer a new research approach to illustrate the material basis of herbal compound.
藿胆丸由广藿香叶和猪胆粉两味组成。藿胆丸源于清代《医宗金鉴》,是治疗慢性鼻炎、慢性鼻窦炎的名优中成药,具有芳香化浊,清热通窍的功效,临床主要用于治疗湿浊内蕴、胆经郁火所致的鼻塞、流清涕、前额头痛等症,已有二百余年的临床应用历史。藿胆丸疗效确切,但产品粗糙,存在生产工艺陈旧、剂型落后、服用量大等不足。本课题开展了如下研究:
1、藿胆丸中广藿香叶有效部位筛选
以抗炎和抗过敏药效试验筛选广藿香叶有效部位,并以成药藿胆丸为对照,结果表明,广藿香叶水溶性部位和不含挥发油的脂溶性部位虽有抗炎作用,但作用强度明显低于挥发油部位、广藿香叶、藿胆丸成药;广藿香叶水溶性部位和不含挥发油的脂溶性部位不具有抗过敏作用,而挥发油部位与广藿香叶、藿胆丸具有抗过敏作用。广藿香叶挥发油同时具有抗炎、抗过敏作用,其作用强度接近广藿香叶、藿胆丸成药,因此可确定藿胆丸中广藿香叶有效部位主要是广藿香挥发油。
比较超临界二氧化碳萃取(以下用SFE-CO_2表示)和水蒸汽蒸馏的方法提取广藿香叶挥发油。结果表明,采用水蒸汽蒸馏,只能得芳香水,甚难得到挥发油。故本课题采用SFE-CO_2萃取技术提取广藿香叶挥发油并确定其提取工艺:广藿香叶粉碎成粗粉,SFE-CO_2提取,提取参数为萃取温度53℃,萃取压力13MPa,解析压力5.5MPa,解析温度63℃,二氧化碳(CO_2)流量为400L/h,萃取时间为2.0小时,得广藿香叶挥发油。
采用SFE-CO_2萃取所得广藿香叶挥发油,与市售广藿香油比较,两者质量一致,均符合2005年版中国药典规定。
2、广藿香油对藿胆丸药效学的影响
用广藿香油替代藿胆丸中的广藿香叶药材所得的样品称为精制藿胆方,即精制藿胆方的组成为广藿香油和猪胆粉。比较精制藿胆方、藿胆丸、广藿香油、猪胆粉的药效学,结果表明:
(1)精制藿胆方、广藿香油、猪胆粉与藿胆丸均具有抗炎作用。抗炎指标有醋酸所致小鼠腹腔毛细血管通透性增高、角叉菜胶所致大鼠足趾肿胀、二甲苯所致小鼠耳廓肿胀,大鼠棉球肉芽肿等四种急性和亚急性炎症模型。精制藿胆方对大鼠角叉菜胶所致足趾肿胀、二甲苯所致小鼠耳廓肿胀等急性炎症模型、大鼠棉球肉芽肿的亚急性炎症模型的抑制作用强度与藿胆丸相当,对抑制醋酸所致小鼠毛细血管通透性增高作用优于藿胆丸。广藿香油、猪胆粉两者配伍后协同增效作用明显。提示用广藿香油替代藿胆丸中的广藿香叶,可以保持藿胆方的抗炎作用。
(2)精制藿胆方、广藿香油、猪胆粉与藿胆丸原方均具有抗过敏作用。抗过敏指标有卵蛋白致敏豚鼠离体回肠肌的过敏性收缩反应,同种大鼠被动皮肤过敏反应(PCA),2,4—二硝基氯苯所致小鼠耳廓皮肤迟发型超敏反应等过敏反应模型。精制藿胆方对卵蛋白致敏豚鼠离体回肠肌的过敏性收缩反应、同种大鼠被动皮肤过敏反应(PCA)的抑制作用优于藿胆丸;对2,4—二硝基氯苯所致小鼠耳廓皮肤迟发型超敏反应的抑制作用强度相当于藿胆丸。广藿香油、猪胆粉两者配伍后协同增效作用明显。提示用广藿香油替代藿胆丸中的广藿香叶,可以保持藿胆方的抗过敏作用。
(3)精制藿胆方具有体内外抗菌作用。精制藿胆方体外抗菌试验对所试菌种(金黄色葡萄球菌、甲型溶血性链球菌、乙型溶血性链球菌、肺炎链球菌、卡他球菌、白喉棒状杆菌、大肠埃希氏菌、绿脓假单胞菌及白色念珠菌)均有不同程度的抗菌作用;对感染肺炎双球菌、乙型溶血性链球菌、金黄色葡萄球菌的小鼠,均具有显著的降低死亡率的作用。提示用广藿香油替代藿胆丸中的广藿香叶,可以保持藿胆方的体内外抗菌作用。
(4)精制藿胆方、广藿香油、猪胆粉与藿胆丸均具有增强免疫作用。增强免疫作用指标有小鼠网状内皮系统碳粒廓清吞噬作用的模型。精制藿胆方、广藿香叶挥发油、猪胆粉均对小鼠网状内皮系统碳粒廓清的吞噬作用具有显著增强作用;精制藿胆方增强免疫作用强度与藿胆丸相当。提示用广藿香油替代藿胆丸中的广藿香叶,可以保持藿胆方的增强免疫作用。
研究结果表明,由广藿香油和猪胆粉组成的精制藿胆方具有抗炎、抗过敏、抗菌、增强免疫作用;广藿香油与猪胆粉两者配伍应用,在抗炎、抗过敏药效中,具有显著的协同增效作用;在抗炎、抗过敏、抗菌、增强免疫各药效指标中,精制藿胆方相当于甚至优于藿胆丸。结果提示,藿胆丸中的广藿香叶以广藿香油替代,可保持原藿胆丸的药效。
3、猪胆粉化学成分对肝功和血液生化指标的影响
由广藿香油和猪胆粉组成的精制藿胆方中,仍以猪胆粉直接入药,从而造成成型困难、气味难闻、卫生学难过关、质量难监控,同时由于猪胆粉含一定量的胆固醇,长期服用可能会引起血中胆固醇偏高等缺陷,迫切需要利用现代科学技术寻找出与其药效等同的有效部位或有效成分取代。
猪去氧胆酸是猪胆粉的主要有效成分,具有化学成分单一,质量稳定可控,同时不含胆固醇、杂质少、易成型等优势。现代药理研究发现,胆酸类成分是猪胆粉多方面药理作用的有效成分,尤其是在抗炎、抗过敏、抗菌及增强免疫方面,胆酸类成分的作用表现更为突出。而猪胆粉中的胆酸类成分主要含猪去氧胆酸,为猪去氧胆酸替代藿胆丸中的猪胆粉提供了理论依据。
用广藿香油和猪去氧胆酸组成藿胆有效成方,即藿胆有效成方的组成为广藿香油和猪去氧胆酸。比较精制藿胆方和藿胆有效成方对血中胆固醇及肝功能损害的影响。结果表明,药后90天,精制藿胆方的血中胆固醇、ALP、AST水平均高于空白对照组;精制藿胆方ALT的水平与空白组相当;停药14天,精制藿胆方血中胆固醇、ALP、AST、ALT水平均恢复正常;藿胆有效成方给药后90天血及停药14天血中胆固醇、ALP、AST、ALT水平均与空白对照组相当。实验结果提示,精制藿胆方长期用药会导致血中胆固醇及部分转氨酶偏高,藿胆有效成方则可避免该结果。猪去氧胆酸替代猪胆粉应用于藿胆丸中,可以避免对血中胆固醇及肝功能损害的影响,使其安全性得到更好的保障。
4、猪去氧胆酸对藿胆丸药效学的影响
藿胆有效成方能够避免对血中胆固醇及肝功能损害的影响,是否能保持藿胆丸的药效尚需实验证实。以抗炎、抗过敏、抗菌及免疫功能试验等指标,来评价猪去氧胆酸的替代是否可行。结果表明:
(1)藿胆有效组分方、猪去氧胆酸、广藿香油、精制藿胆方、猪胆粉及藿胆丸原方均对醋酸所致小鼠腹腔毛细血管通透性增高、二甲苯所致小鼠耳廓肿胀,角叉菜胶所致大鼠足趾肿胀、大鼠棉球肉芽肿均有明显抑制作用。藿胆有效组分方对小鼠腹腔毛细血管通透性增高、二甲苯所致小鼠耳廓肿胀急性炎症模型的抑制作用与藿胆丸原方、精制藿胆方相当。藿胆有效组分方对角叉菜胶所致大鼠足趾肿胀、大鼠棉球肉芽肿的抑制作用略优于藿胆丸原方及精制藿胆方。猪去氧胆酸对醋酸所致小鼠腹腔毛细血管通透性增高、二甲苯所致小鼠耳廓肿胀,角叉菜胶致大鼠足趾肿胀、大鼠棉球肉芽肿的抑制作用与猪胆粉相当。猪去氧胆酸、广藿香油单独应用均有较好的抗炎作用,且两药配伍有协同增效的抗炎作用。实验结果提示,用猪去氧胆酸替代藿胆丸中的猪胆粉、可以保持藿胆方的抗炎作用。
(2)藿胆有效组分方、猪去氧胆酸、广藿香油、精制藿胆方、猪胆粉及藿胆丸原方均对卵蛋白致敏豚鼠离体回肠肌的过敏性收缩反应,具有快速抑制作用;对同种大鼠被动皮肤过敏反应(PCA)具有明显的抑制作用;对2,4—二硝基氯苯所致小鼠耳廓皮肤迟发型超敏反应有明显的抑制作用。藿胆有效组分方对卵蛋白致敏豚鼠离体回肠肌的过敏性收缩反应,对同种大鼠被动皮肤过敏反应(PCA),对2,4—二硝基氯苯所致小鼠耳廓皮肤迟发型超敏反应的抑制作用强度与精制藿胆方、藿胆丸原方相当。猪去氧胆酸对卵蛋白致敏豚鼠离体回肠肌的过敏性收缩反应,对同种大鼠被动皮肤过敏反应(PCA),对2,4—二硝基氯苯所致小鼠耳廓皮肤迟发型超敏反应的抑制作用强度与猪胆粉相当。猪去氧胆酸、广藿香油单独应用均有较好的抗过敏作用,且两药配伍有协同增效的抗过敏作用。实验结果提示,用猪去氧胆酸替代藿胆丸中的猪胆粉、可以保持藿胆方的抗过敏作用。
(3)藿胆有效组分方、猪去氧胆酸、广藿香油、精制藿胆方、猪胆粉及藿胆丸原方均能显著增强小鼠网状内皮系统碳粒廓清的吞噬作用,藿胆有效组分方对小鼠网状内皮系统碳粒廓清的吞噬功能的作用强度与藿胆丸原方、精制藿胆方相当。猪去氧胆酸对小鼠网状内皮系统碳粒廓清的吞噬功能的作用强度与猪胆粉相当。猪去氧胆酸、广藿香油单独应用均有较好的增强免疫功能的作用,且两药配伍有协同增效的增强免疫功能的作用。实验结果提示,用猪去氧胆酸替代藿胆丸中的猪胆粉、可以保持藿胆方的增强免疫功能的作用。
(4)藿胆有效组分方、精制藿胆方体外抗菌试验对所试菌种(金黄色葡萄球菌、甲型溶血性链球菌、乙型溶血性链球菌、肺炎链球菌、卡他球菌、白喉棒状杆菌、大肠埃希氏菌、绿脓假单胞菌及白色念珠菌)的MIC,藿胆有效组分方介于9.8×10-5g/mL～2.0×10-1g/mL之间,精制藿胆方介于6.2×10-3g/mL～2.0×10-1g/mL之间,提示藿胆有效组分方体外抗菌作用强度比精制藿胆方强。藿胆有效组分方、精制藿胆方对感染肺炎双球菌混悬液、乙型溶血性链球菌混悬液、金黄色葡萄球菌混悬液的小鼠的死亡率有不同程度的抑制作用,藿胆有效组分方对感染肺炎双球菌混悬液、乙型溶血性链球菌混悬液、金黄色葡萄球菌混悬液的小鼠的死亡率的抑制作用明显优于精制藿胆方。提示猪去氧胆酸替代藿胆丸中的猪胆粉,更有利于其抗菌作用的发挥。
通过对藿胆丸拆方研究广藿香叶和猪胆粉的有效部位或有效成分,并与藿胆丸原方进行比较,明确了藿胆丸原方产生临床疗效的有效部位是由广藿香油和猪去氧胆酸,且二者之间具有协同增效的作用。采用藿胆丸有效成分组方不但可以避免藿胆丸原方的不良反应,而且从根本上克服传统藿胆丸剂量大、成型难,成品质量波幅大等技术问题,提高了藿胆丸的技术含量,为产品质量达到“安全、有效、稳定、可控”提供技术支撑。
本研究明确了藿胆丸治疗鼻炎的物质基础,用广藿香油和猪去氧胆酸组成的处方,具有药效作用物质明确,能够达到现代化中成药的雏形,为藿胆丸的产品升级换代奠定理论基础,提供了科学的依据。本研究方法可作为阐明中药复方物质基础提供一种科学的研究思路和方法。
Studying on the substance foundation of Chinese Herb Pharmacodynamic is the basis of research and development a new generation Chinese Patent Medicine with definite effective components,exact efficacy,stable and controllable quality,also the key to achieve modernization of Chinese traditional medicine.To investigate the substance foundation of Huodan pill by means of the Purification & Recombinant is the innovation in this project. According to the chemical composition and the feature in this formula,we could apply modern industrial technology to get various chemical compositions of Herba Pogostemonis' leaf and Porcine Gall Powder,and verify the active components by tracing and analysis the anti-inflammatory and anti-allergic chemical compositions with reference to traditional pill, which were used as compounds to Investigation on compatibility and dose optimization.It can lay a foundation of previous research for developing a new traditional Chinese medicine.
Huodan pill,rooted from Yi Zong Jin Jian in Qing dynasty,is a famous Chinese Patent Medicine treating chronic rhinitis and chronic sinusitis.With the efficacy of resolving turbidity with aroma,clearing heat and relieving stuffiness,it can be used to treating intrinsic damp turbidity,nasal obstruction,nasal discharge and prefrontal ache accused by stagnant fire in Dan channel.It has a clinical application history more than two hundred years.Huodan pill is consist of two parts:Herba Pogostemonis' leaf and Porcine Gall Powder.Although the definite therapeutic effect,there are a large of disadvantages such as the rough product,old production process,backward dosage forms.The project developed a series of research as below:
1 Screening of active part ofHerba Pogostemonis' leaf in Huodan pill. We screened active part of Herba Pogostemonis' leaf by pharmacodynamic test used in anti-inflammation and anti-hypersensitiveness drug,and used patent medicine Huodan pill as the control group.The results demonstrated that hydrophilic and lipophilic part without volatile oil of Herba Pogostemonis' leaf have the effect of anti-inflammation,but lower than volatile oil,total extraction of Herba Pogostemonis' leaf and the control group.At the same time,both parts have little effect of anti-hypersensitiveness.While volatile oil of Herba Pogostemonis' leaf have the same anti-inflammation effect as total extraction of Herba Pogostemonis' leaf and the control group,as well as the anti-hypersensitiveness effect.We concluded that volatile oil is the main active part of Herba Pogostemonis' leaf in Huodan pill.
We extracted volatile oil from Herba Pogostemonis' leaf by two methods:SFE-CO2 and distillation,and the results showed that only aromatic water can be gotten by the method of distillation.So the SFE-CO_2 is adopted to extract volatile oil.The protocol of extraction is the following:Herba Pogostemonis' leaf is crashed to coarse powder first,and then extracted by SFE-CO_2.The extract temperature is 53℃,extract pressure is 13MPa,while the resolve pressure is 5.5Mpa,the resolve temperature is 63℃,the flow rate of CO_2 is 400L/h.The whole process lasted 2 hours.And then the volatile oil can be gotten. Volatile oil extracted by SFE-CO_2 accord to Herba Pogostemonis' oil in market,and both are consistent with Chinese Pharmacopeia.
2 Evaluation ofHerba Pogostemonis' oil in Huodan pill. Refined HuoDan Recipe is comprised ofHerba Pogostemonis' oil and Porcine Gall Powder. We studied pharmacological effect of the refined HuoDan Recipe,Huodan pill,Herba Pogostemonis' oil,Porcine Gall Powder and results as showed below.
a) Refined Huodan Recipe,Herba Pogostemonis' oil,Porcine Gall Powder and Huodan pill all have anti-inflammatory effects.The indexes include increasing capillary permeability of mouse belly cavity,swelling of rat toes by carrageenan, swelling of mouse auricle by dimethl benzene and rat cotton ball granuloma.,the acute and subacute inflammation model.Refined Huodan Recipe has the same inhibitory effect on the subacute inflammatory model such as swelling of rat toes by carrageenan,swelling of mouse auricle by dimethl benzene and rat cotton ball granuloma as Huodan pill,and better inhibitory effect on increasing capillary permeability of mouse belly cavity by acetic acid than Huodan pill.Herba Pogostemonis' oil and Porcine Gall Powder,synergize the anti-inflammatory effect, which implies that refined Huodan recipe has good anti-inflammatory effect as well as compositive materials,and which have synergism.
b) There are all anti-allergy effect in Refined Huodan Recipe,Herba Pogostemonis'oil,Porcine Gall Powder and original prescription of Huodan pill. The indexes include allergia contractile response of ileum muscle in cobaya by egg albumin,passive cutaneous ana-phylaxis reaction(PCA),delayed hypersensitivity of mouse's pinna skin by 2,4-dinitro-chlorobenzene.Refined Huodan Recipe has higher depressant effect on Allergic Contractile response of cony pig by Ovalbumin and PCA of same species of rat than Huodan pill.The results have indicated that Refined Huodan Recipe and its disassembled prescriptions have remarkable anti-allergy effects.Drug synergy has the relationship of synergistic action.
c) Refined Huodan Recipe has significant anti-microbial doughty effects.Refined Huodan Recipe has obvious dispatity degree anti-bacterial action against staphylococcus aureus,alpha hemolytic streptococcus,beta-hemolytic streptococci, streptococcus pneumoniae,micrococcus catarrhalis,corynebacterium diphtheriae, escherichia,aeruginosus bacillus,blastomyces albicans;The in corpore anti-microbial tests showed that Refined Huodan recipe has doughty inhibit activity to diplococcus lanceolatus,staphylococcus aureus,beta-hem olytic streptococci.These research results indicate that Refined Huodan recipe has anti-microbial doughty effects.
d) Refined Huodan Recipe,Herba Pogostemonis'oil,Porcine Gall Powder and Huodan pill have significant enhancing immunity function effects.Drug synergy has the relationship of synergistic action.Refined Huodan Recipe,Herba Pogostemonis'oil,Porcine Gall Powder can obvious strengthen the phagocytosis of mice's reticuloendothelial system.Refined Huodan Recipe's enhancing immunity function effect is as the same as Huodan pill.The results have indicated that Refined Huodan Recipe and its disassembled prescriptions have remarkable enhancing immunity function effects.
The results of the experiment indicated that Refined Huodan Recipe,which made of Herba Pogostemonis'oil and Porcine Gall Powder,havs notable anti-inflammatory effects, anti-allergy effects,anti-microbial effects and enhancing immunity function effects.Drug synergy has the relationship of synergistic action.The results show that substitute Herba Pogostemonis'oil for Herba Pogostemonis' leaf,Refined Huodan Recipe can reserve its effect.
3 Effect of Porcine Gall Powder composition for liver function and blood biochemical. Because of Porcine Gall Powder,the Huodan Pills have the disadvantage of difficult forming,the older preparation,large taken dose,foul smells and difficult controlling of the microorganism.In addition,Because of the amount of cholesterol and Chenodeoxycholic acid in Porcine Gall Powder,you will have cell toxicity,abnormal determination of cholesterol in your blood and liver function damage,if you take in a long time of Huodan pills.In order to explore a kind of preparation that has the advantage of defined effective medium,stable quality,well-defined Pharmacological effect,well-defined therapy and normal determination of cholesterol in your blood,although you take in a long time of them. It is very necessary to look for the active site or the effective component of the Porcine Gall Powder,which can Substitute of Porcine Gall Powder Using in Huodan Pills.The Cholic acid has many kinds of Pharmacological effect,especially in anti-inflammatory,anti-allergy, anti-microbial and enhancing immunity function effects.The other Cholic acid is very little, no notion of Pharmacological effect and some are hurt to liver function and cardiovascular system.
Hyodeoxycholic acid is the main active component of Porcine Gall Powder,which has testified by modern research.It has the advantage of one chemical compound,stable quality, clearing Pharmacological effect,no foreign matter and no the amount of cholesterol and easy forming,which offers the theoretical support to Hyodeoxycholic acid substitute of Porcine Gall Powder using in Houdan pills.
Porcine Gall Powder recipe can increase the level of cholesterol in blood and reduce the damage of the liver function,but Hyodeoxycholic acid recipe can not,if you take in a long time of them.The levels of cholesterol,AST、ATP in blood of the Porcine Gall Powder recipe low,middle,high dose groups is higher than the blank group,the level of ALT as the same as the blank group,after giving drug for 90 days.The levels of cholesterol,AST,ATP in blood of the Porcine Gall Powder recipe low,middle,high dose groups come back into normal level after stopping using drug for 14 days.The levels of cholesterol,AST、ATP、ALT in blood of the Hyodeoxycholic acid recipe low,middle,high dose group is the same as the blank group,after giving drug for 90 days and stopping using drug for 14 days. The results of the experiment indicate that using Hyodeoxycholic acid Substituting of Porcine Gall Powder Using in Huodan Pills makes to raise the safety of Huodan Pills.
4 Effect of Hyodeoxycholic acid for the pharmacodynamics of Huodan pill This effective compound recipe consists of Herba Pogostemonis'oil and Hyodeoxycholic acid could avoid from injuring the liver,but it can be still effective as Huodan pill? Four kinds evaluating indicator of rhinitis such as anti-inflammatory,anti-allergy,anti-bacterial and enhancing immunity function experiment were used to compare the pharmacology of the Hyodeoxycholic acid instuting ofPulvis Fellis Suis before and after.The conclusion is:
a) Huodan effective component recipe,Hyodeoxycholic acid,Herba Pogostemonis'oil, Refined Huodan Recipe,Porcine Gall Powder and Huodan pill has significant anti-inflammatory effects.Hyodeoxycholic acid and Oleum pogostemonis have the relationship of synergistic action.Hyodeoxycholic acid recipe has the same inhibitory action on mice's auricle swelling induced by xylene,the increase of capillary permeability in mice induced by acetic acid,rat's toes swelling induced by carrageenan, the granuloma by the cotton ball as Huodan pills and Porcine Gall Powder recipe. Hyodeoxycholic acid has the same inhibitory action on mice's auricle swelling induced by xylene,the increase of capillary permeability in mice induced by acetic acid,rat's toes swelling induced by carrageenan,the granuloma by the cotton ball as Porcine Gall Powder.Hyodeoxycholic acid and Oleum pogostemonis have significant anti-inflammatory effects.Drug synergy has the relationship of synergistic action.The results of the experiment indicate that Hyodeoxycholic acid is the active component of the Porcine Gall Powder in anti-inflammatory pharmacological effects and Hyodeoxycholic acid may be institute of Porcine Gall Powder using in Huodan Pills.
b) Huodan effective component recipe,Hyodeoxycholic acid,Herba Pogostemonis'oil, Refined Huodan Recipe,Porcine Gall Powder and Huodan pill has the relationship of synergistic action.Hyodeoxycholic acid,Oleum pogostemonis and Huodan recipe have obvious inhibitory action on extracorporeal ileum muscles' hypersensitive contractility reaction of Guinea pig sensitized by albumin,homogeneous passive cutaneous anaphylaxis in mice,mice's auricle cutaneous forme tardive supersensitivity action by 2,4-dinitrochlorobenzene.The results have indicated that Huodan recipe and its disassembled prescriptions have remarkable anti-allergy effects.Drug synergy has the relationship of synergistic action.
c) Huodan effective component recipe,Hyodeoxycholic acid,Herba Pogostemonis'oil, Refined Huodan Recipe,Porcine Gall Powder and Huodan pill have significant enhancing immunity function effects.Drug synergy has the relationship of synergistic action.Hyodeoxycholic acid,Oleum pogostemonis and Huodan recipe can obvious strengthen the phagocytosis of mice's reticuloendothelial system.The results have indicated that Huodan recipe and its disassembled prescriptions have remarkable enhancing immunity function effects.Drug synergy has the relationship of synergistic action.
d) Huodan effective component recipe,Hyodeoxycholic acid,Herba Pogostemonis'oil, Refined Huodan Recipe,Porcine Gall Powder and Huodan pill has significant anti-microbial doughty effects.Huodan recipe has obvious dispatity degree anti-bacterial action against staphylococcus aureus,alpha hemolytic streptococcus, beta-hemolytic streptococci,streptococcus pneumoniae,micrococcus catarrhalis, corynebacterium diphtheriae,escherichia,aeruginosus bacillus,blastomyces albicans; The in corpore anti-microbial tests showed that refined huodan recipe has doughty inhibit activity to diplococcus lanceolatus,staphylococcus aureus,beta-hem olytic streptococci.Huodan recipe has anti-microbial doughty effects.
By studying on the effective fractions or components of Herba Pogostemonis' leaf and Porcine Gall Powder respectively,and comparing with the original formula,it could affirm that the effective parts are patchouli oil and hyodeoxycholic acid which have the synergistic interaction.Formulating with the effective compositions of Huodan pill not only could avoid the adverse reaction of the original prescription,but also overcoe the technical problems fundamentally such as large dosage,hard shaping and unstable quality.It offered a technological support for the high quality product of "safe,effective,stable,and controllable".
This project has made clear the material basis of treating rhinitis.The formula consists of patchouli oil and hyodeoxycholic acid is a rudiment of the modern Chinese patent medicine. It provided scientific basis and laid a theoretical foundation for product renewal.This study method could offer a new research approach to illustrate the material basis of herbal compound.
引文
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