广痛消有效成分含量测定及健康人体药代动力学研究
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摘要
研究背景及目的意义
背景：肛肠科治疗的疾病主要包括：肛瘘、痔疮、直肠脱垂、直肠肛管周围脓肿、直肠癌、肛裂等,这些疾病通常需要通过手术才能达到根治的目的,但手术后常会出现疼痛、水肿、出血、感染、创面延迟愈合等并发症,针对这些并发症除针对性治疗外,术后换药成为治疗这些术后并发症、促进创面愈合的重要手段。目前临床上常用的剂型一般为痔疮膏、痔疮栓之类,这些传统的药物剂型通过肛门给药,直接作用于手术创面或直肠黏膜发挥抗炎、镇痛、消肿等作用,但也同时存在与病变部位接触面小、分布不均匀、易融化排出、药效持续时间短、生物利用度低等缺点。正是基于目前肛肠疾病术后换药所用剂型受限的现状,赵宝明教授根据中医基础理论结合肛肠疾病自身的发病特点研制出一种中药泡沫气雾剂——广痛消泡沫气雾剂,它除具备抗炎、消肿、解痉镇痛等功效外,还具有喷出物不流动,泡沫稳定、持续时间长,药物在病灶部位作用时间长,涂布面广,均匀分散在腔道内,药物能有效地渗入黏膜皱襞等优点,其疗效优于其他剂型,是一种极具研发价值的剂型。
“广痛消”中的”广”即广肠,包括乙状结肠和直肠,如《证治要诀》云”广肠,言其广阔于大小肠也”,表明了广痛消泡沫气雾剂可以作用的部位；”痛消”阐明该药有镇痛功效；“泡沫气雾剂”点明了该药的剂型。“广痛消泡沫气雾剂”精炼地概括了该中药复方的使用部位、疗效和剂型。
广痛消泡沫气雾剂先后在广安门医院肛肠科及实验室、东直门医院肛肠科及中心实验室、北京中医药大学中药学院、军事科学院和北医病理系的协助下完成了该药的提取工艺、药物质量标准、药效、剂型优势和药物安全性等方面的研究。其动物实验和临床研究历时20余年,有着深厚的研究基础。
目的：尽管目前已完成了该泡沫气雾剂部分药效的研究,但尚存在药物在体内吸收、代谢的情况不明确,其发挥作用的机理不清楚等情况,故本课题通过测定延胡索、当归、厚朴、细辛、白芍、黄柏、茜草的有效成分含量及健康人体的药代动力学研究以期对阐述其组方原理,对新药开发,质量评价及指导临床合理用药提供参考,也为今后该泡沫气雾剂的机制研究奠定基础。
实验前期曾采用高效液相色谱法检测广痛消水提物中有效成分,但存在提取物的药效成分相互干扰严重,无法进行准确的定量分析；一些有效成分含量低,无明显的响应信号；操作繁琐、灵敏度低、分析速度慢、多成分含量同时测定色谱条件摸索困难等问题,故改用液质联用(LC-MS/MS)。液质联用技术是以液相色谱作为分离系统,质谱为监测系统,样品在质谱部分和流动相分离,被离子化后,经质谱的质量分析器将离子碎片按质量数分开,经检测器得到质谱图。液质联用体现了色谱和质谱的优势互补,将色谱对复杂样品的高分离能力,与MS具有高选择性、高灵敏度及能够提供相对分子质量与结构信息的优点结合起来,在药物分析、食品分析和环境分析等许多领域得到了广泛应用。
研究内容及结果
一、广痛消水提物中5种有效成分的含量的测定
目的：建立灵敏、快速、简便的液相色谱-串联质谱法测定广痛消水提物中主要有效成分(延胡索乙素、芍药苷、盐酸小檗碱、厚朴酚、和厚朴酚)的含量。
方法：采用色谱柱：Agilent Eclipse Plus C18柱(100mm×2.1mm,3.5μm),流动相A：0.1%甲酸水,B：乙腈,梯度洗脱,柱温30℃。质谱条件：ESI离子源,正、负离子检测方式,干燥气温度300℃,干燥气体流量10L·min-1；雾化器压力30Pa；毛细管电压正离子4000V,负离子3500V,用于定量分析的离子反应m/z分别为芍药苷(m/z479.2→m/z121)、延胡索乙素(m/z356.2→m/z192.2)、盐酸小檗碱(m/z336.1→m/z320.2)、厚朴酚(m/z265.1→m/z247.1)、和厚朴酚(m/z265.2→m/z233.1)、内标布洛芬、消旋山莨菪碱监测离子对分别为m/z205.1→m/z161.1,m/z306.1→m/z140。
结果：芍药苷、延胡索乙素、盐酸小檗碱、厚朴酚、和厚朴酚均得到精确的分析,线性范围分别为：0.8828125-113(r=0.9998)；1.54375-49.4(r=0.9995)；0.9046875-115.8(r=0.9997)；0.88125-112.8(r=0.9994)；0.165625-21.2(r--0.9999)。加样回收率分别为1.998%,1.14%,3.117%,1.828%,1.596%。
结论：该方法灵敏度高、分析速度快、定性能力强、定量能力好,可同时测定广痛消水提物中5种主要有效成分含量。
二、广痛消泡沫气雾剂直肠给药的健康人体药代动力学研究
目的：建立快速、灵敏的液相色谱-串联质谱(LC-MS/MS)法直接测定健康人体血浆中延胡索乙素、盐酸小檗碱的含量,求取药动学参数。
方法：血浆样品以丙酮沉淀蛋白,80%乙腈萃取,对广痛消水提物中5种有效成分含量测定的色谱、质谱条件进行微调,色谱柱：Agilent Eclipse Plus C18柱(100mm×2.1mm,3.5μm),流动相A：0.1%甲酸水,B：乙腈,0～lmin(B15%、0.2mL/min)；1～2min(B15%～40%,0.3mL/min);2～7min(B40%～80%,0.3mL);7～8min (B80%～15%,0.3mL/min);8～10min(B15%,0.3mL/min);10～11min(B15%,0.2mL/min);12min(B15%,0.2mL/min)质谱条件：ESI离子源,正离子检测方式,干燥气温度300℃,干燥气体流量10L·min-1；雾化器压力30Pa；毛细管电压正离子4000V,监测离子对分别为盐酸盐酸小檗碱(m/z479.2→m/z121),延胡索乙素(m/z356.2→m/z192.2),内标消旋山莨菪碱(m/z306.1→m/zl40),自动进样盘温度15℃,柱温30℃,并进行了系统的方法学评价。
结果：延胡索乙素和盐酸小檗碱在各自的生物样品浓度范围内,均呈良好线性,r>0.99；其各成分的线性范围分为：0.000594-1.188ng/mL,0.000536-1.072ng/mL,最低定量限为：0.000536ng/ml,日内、日间精密度均小于15%,提取回收率均大于85%,8名健康志愿者肛门给广痛消泡沫气雾剂后,延胡索乙素、盐酸小檗碱的主要药代动力学参数Tmax分别为：132.5±25.4951min和132.5±30.11881min,Cmax分别为：0.550738±0.300594ng/ml和0.3108±0.324596ng/ml, T1/2分别为69.37085±72.60998min和57.06471±39.74188min, AUClast分别为46.2976±27.85874min*ng/mL和28.25056±28.34036min*ng/mL,
结论：该方法操作简单,效率高,选择性好,灵敏度高,可成功用于广痛消泡沫气雾剂的健康人体血浆的动力学研究。
Research background and objective
Department of anorectal surgery usually treat diseases mainly include:anal fistula, hemorrhoids, rectum prolapse, rectum and anal canal abscess, rectal cancer, anal fissure, etc. These diseases usually need surgical treatment, But the pain, edema, hemorrhage, infection, delayed wound healing complications often appear after surgical treatments. The postoperative dressing changes become an important means to treat these postoperative complications and promote wound healing in addition to symptomatic treatments.At present, hemorrhoids ointment and hemorrhoids suppository are widely used to treat anorectal diseases, These drugs are supplied through the anus,They play an anti-inflammatory, analgesic, swelling by direct role in the surgical wound or the rectum mucosa. While there are small contact surface at the lesion site, uneven distribution, easy to melt and discharge, short duration of efficacy, low bioavailability. These are based on the above situation, Professor Zhao Baoming developed Guang TongXiao foam aerosol that based on the basic theory of TCM and combined with anorectal disease clinical characteristics,the foam aerosol can not only anti-inflammatory, swelling, relieve spasm and pain but also following advantages:the ejecta no flow, foam stable, long duration in the lesion site, coated with a wide range of uniformly dispersed in the lumen, effectively penetrate the mucosal folds, etc. It's more effective than other formulations,and is a great development potential of dosage form.
GuangTongXiao foam aerosols is a kind of traditional Chinese medicine foam aerosols. Professor Zhao Baoming developed this foam aerosols that is based on the basic theory of TCM and combined with characteristics of anorectal diseases. It is consisted of corydalis, white peony root, rubia cordifolia and othe Chinese medicines, it has spasmolysis analgesia, anti-inflammatory, hemostasis and promote wound healing. Treatment tips say"guang", means small and large intestines;"Tongxiao" means this foam aerosols have analgesia efficacy. Foam aerosol illustrate the formulations of the herbal compound."GuangTongXiao foam aerosols" summed the site of action of the herbal compound, efficacy and dosage forms.The foam aerosol has been finished these researches such as the extraction process、quality standards. efficacy, dosage form advantages and drug security researches by the department of Anorectal Surgery and laboratory of Guang An Men Hospital, the Department of Anorectal Surgery and the central laboratory of Dongzhimen Hospital、 Beijing University of Chinese Medicine the Academy of Military Sciences and Department of Pathology of Peking University Health Science. It has strong research bases because the animal experiments and clinical studies lasted more than20years, but these efficacy reasons unclear, Therefore,this subject will lay the foundation for the future study of the mechanism and guide clinical medication by determination of the active ingredient contents and pharmacokinetic study of healthy volunteers.
Research methods and contents
1.To determine five effective components in the water extract of GuangTongXiao.In this study, using LC-MS/MS method determine simultaneously tetrahydropalmatine, paeoniflorin, berberine, magnolol and honokiol, The results show that The method is rapid and sensitive, attribute strong, This method can determine simultaneously five kinds of effective components. LC-MS/MS method for determining contents of tetrahydropalmatine, paeoniflorin, berberine, magnolol and honokiol, the ibuprofen, raceanisodamine act internal standard. Chromatographic conditions:chromatographic column(Agilent Eclipse Plus C18,100mm×2.1mm,3.5μm), mobile phase A:0.1%of formic acid, B:acetonitrile, gradient elution, the column temperature30℃. Mass spectrometer conditions:ESI ion source, the positive ion mode drying gas temperature300℃, gas flow10L-min-1; Nebulizer pressure30Pa; the capillary voltage positive ions4000V, Paeoniflorin tetrahydropalmatine berberine, magnolol, honokiol, ibuprofen and raceanisodamine monitoring of ion-pair479.2/121,356.2/192.2,336.1/320.2,265.1/247.1,265.2/233.1,205.1/161.1,306.1/140,they are all analysised accurately.Linear ranges:0.8828125-113(r=0.9998);1.54375-49.4(r=0.9995);0.9046875-115.8(r=0.9997);0.88125-112.8(r=0.9994);0.165625-21.2(r=0.9999). spike recovery1.998%,1.14%,3.117%,1.828%and1.596%.The results show that the method high sensitivity, fast analysis,while qualitative ability and quantitative ability good, the method can determine simultaneously five main active ingredient contents in GuangTongXiao aqueous extracts.
2. To study pharmacokinetics of Guangtongxiao foam aerosols that act on healthy human.The health human pharmacokinetics of GuangTongXiao foam aerosols that is administrated by anal.Determining the concentrations of tetrahydropalmatine and berberine in healthy male plasma by LC-MS/MS, raceanisodamine act internal mark, protein of plasma samples was precipitated by acetone,80%acetonitrile extraction, Chromatographic conditions:chromatographic column(Agilent Eclipse Plus C18,100mm×2.1mm,3.5μm), mobile phase A:0.1%of formic acid, B:acetonitrile, gradient elution0～1min(B15%、0.2mL/min);1～2min(B15%～40%,0.3mL/min);2～7min(B40%～80%,0.3mL);7～8min (B80%～15%,0.3mL/min);8～10min(B15%,0.3mL/min);10llmin(B15%,0.2mL/min);12min (B15%、0.2mL/min) Mass spectrometer conditions: ESI ion source, the positive ion mode drying gas temperature300℃, gas flow10L-min-1; Nebulizer pressure30Pa; the capillary voltage positive ions4000V, monitoring of ion-pair are479.2/121,356.2/192.2,306.1/140, the autosampler temperature15℃, the column temperature30℃. methodology evaluation show the tetrahydropalmatine and berberine hydrochloride that in biological samples show a good linear r>0.99, The linear range of the respective components are0.000594-1.188ng/mL,0.000536-1.072ng/mL, Lowest limit of quantification is0.000536ng/ml, intraday precision and day precision are both less than 15%,The extraction recovery are both more than85%. The method is simple, high efficiency, good selectivity, high sensitivity, and can be used successfully in the study of the dynamics of human plasma thay contain GuangTongXiao foam aerosols, On this basis, a high sensitivity, specificity, and accurate LC-MS/MS method has been establed, two kinds of active ingredients in the foam aerosols are determined simultaneously, and research the dynamic characteristics of foam aerosol in healthy male body,in order to provide a reference for the clinical and the late further researches.
背景：肛肠科治疗的疾病主要包括：肛瘘、痔疮、直肠脱垂、直肠肛管周围脓肿、直肠癌、肛裂等,这些疾病通常需要通过手术才能达到根治的目的,但手术后常会出现疼痛、水肿、出血、感染、创面延迟愈合等并发症,针对这些并发症除针对性治疗外,术后换药成为治疗这些术后并发症、促进创面愈合的重要手段。目前临床上常用的剂型一般为痔疮膏、痔疮栓之类,这些传统的药物剂型通过肛门给药,直接作用于手术创面或直肠黏膜发挥抗炎、镇痛、消肿等作用,但也同时存在与病变部位接触面小、分布不均匀、易融化排出、药效持续时间短、生物利用度低等缺点。正是基于目前肛肠疾病术后换药所用剂型受限的现状,赵宝明教授根据中医基础理论结合肛肠疾病自身的发病特点研制出一种中药泡沫气雾剂——广痛消泡沫气雾剂,它除具备抗炎、消肿、解痉镇痛等功效外,还具有喷出物不流动,泡沫稳定、持续时间长,药物在病灶部位作用时间长,涂布面广,均匀分散在腔道内,药物能有效地渗入黏膜皱襞等优点,其疗效优于其他剂型,是一种极具研发价值的剂型。
“广痛消”中的”广”即广肠,包括乙状结肠和直肠,如《证治要诀》云”广肠,言其广阔于大小肠也”,表明了广痛消泡沫气雾剂可以作用的部位；”痛消”阐明该药有镇痛功效；“泡沫气雾剂”点明了该药的剂型。“广痛消泡沫气雾剂”精炼地概括了该中药复方的使用部位、疗效和剂型。
广痛消泡沫气雾剂先后在广安门医院肛肠科及实验室、东直门医院肛肠科及中心实验室、北京中医药大学中药学院、军事科学院和北医病理系的协助下完成了该药的提取工艺、药物质量标准、药效、剂型优势和药物安全性等方面的研究。其动物实验和临床研究历时20余年,有着深厚的研究基础。
目的：尽管目前已完成了该泡沫气雾剂部分药效的研究,但尚存在药物在体内吸收、代谢的情况不明确,其发挥作用的机理不清楚等情况,故本课题通过测定延胡索、当归、厚朴、细辛、白芍、黄柏、茜草的有效成分含量及健康人体的药代动力学研究以期对阐述其组方原理,对新药开发,质量评价及指导临床合理用药提供参考,也为今后该泡沫气雾剂的机制研究奠定基础。
实验前期曾采用高效液相色谱法检测广痛消水提物中有效成分,但存在提取物的药效成分相互干扰严重,无法进行准确的定量分析；一些有效成分含量低,无明显的响应信号；操作繁琐、灵敏度低、分析速度慢、多成分含量同时测定色谱条件摸索困难等问题,故改用液质联用(LC-MS/MS)。液质联用技术是以液相色谱作为分离系统,质谱为监测系统,样品在质谱部分和流动相分离,被离子化后,经质谱的质量分析器将离子碎片按质量数分开,经检测器得到质谱图。液质联用体现了色谱和质谱的优势互补,将色谱对复杂样品的高分离能力,与MS具有高选择性、高灵敏度及能够提供相对分子质量与结构信息的优点结合起来,在药物分析、食品分析和环境分析等许多领域得到了广泛应用。
研究内容及结果
一、广痛消水提物中5种有效成分的含量的测定
目的：建立灵敏、快速、简便的液相色谱-串联质谱法测定广痛消水提物中主要有效成分(延胡索乙素、芍药苷、盐酸小檗碱、厚朴酚、和厚朴酚)的含量。
方法：采用色谱柱：Agilent Eclipse Plus C18柱(100mm×2.1mm,3.5μm),流动相A：0.1%甲酸水,B：乙腈,梯度洗脱,柱温30℃。质谱条件：ESI离子源,正、负离子检测方式,干燥气温度300℃,干燥气体流量10L·min-1；雾化器压力30Pa；毛细管电压正离子4000V,负离子3500V,用于定量分析的离子反应m/z分别为芍药苷(m/z479.2→m/z121)、延胡索乙素(m/z356.2→m/z192.2)、盐酸小檗碱(m/z336.1→m/z320.2)、厚朴酚(m/z265.1→m/z247.1)、和厚朴酚(m/z265.2→m/z233.1)、内标布洛芬、消旋山莨菪碱监测离子对分别为m/z205.1→m/z161.1,m/z306.1→m/z140。
结果：芍药苷、延胡索乙素、盐酸小檗碱、厚朴酚、和厚朴酚均得到精确的分析,线性范围分别为：0.8828125-113(r=0.9998)；1.54375-49.4(r=0.9995)；0.9046875-115.8(r=0.9997)；0.88125-112.8(r=0.9994)；0.165625-21.2(r--0.9999)。加样回收率分别为1.998%,1.14%,3.117%,1.828%,1.596%。
结论：该方法灵敏度高、分析速度快、定性能力强、定量能力好,可同时测定广痛消水提物中5种主要有效成分含量。
二、广痛消泡沫气雾剂直肠给药的健康人体药代动力学研究
目的：建立快速、灵敏的液相色谱-串联质谱(LC-MS/MS)法直接测定健康人体血浆中延胡索乙素、盐酸小檗碱的含量,求取药动学参数。
方法：血浆样品以丙酮沉淀蛋白,80%乙腈萃取,对广痛消水提物中5种有效成分含量测定的色谱、质谱条件进行微调,色谱柱：Agilent Eclipse Plus C18柱(100mm×2.1mm,3.5μm),流动相A：0.1%甲酸水,B：乙腈,0～lmin(B15%、0.2mL/min)；1～2min(B15%～40%,0.3mL/min);2～7min(B40%～80%,0.3mL);7～8min (B80%～15%,0.3mL/min);8～10min(B15%,0.3mL/min);10～11min(B15%,0.2mL/min);12min(B15%,0.2mL/min)质谱条件：ESI离子源,正离子检测方式,干燥气温度300℃,干燥气体流量10L·min-1；雾化器压力30Pa；毛细管电压正离子4000V,监测离子对分别为盐酸盐酸小檗碱(m/z479.2→m/z121),延胡索乙素(m/z356.2→m/z192.2),内标消旋山莨菪碱(m/z306.1→m/zl40),自动进样盘温度15℃,柱温30℃,并进行了系统的方法学评价。
结果：延胡索乙素和盐酸小檗碱在各自的生物样品浓度范围内,均呈良好线性,r>0.99；其各成分的线性范围分为：0.000594-1.188ng/mL,0.000536-1.072ng/mL,最低定量限为：0.000536ng/ml,日内、日间精密度均小于15%,提取回收率均大于85%,8名健康志愿者肛门给广痛消泡沫气雾剂后,延胡索乙素、盐酸小檗碱的主要药代动力学参数Tmax分别为：132.5±25.4951min和132.5±30.11881min,Cmax分别为：0.550738±0.300594ng/ml和0.3108±0.324596ng/ml, T1/2分别为69.37085±72.60998min和57.06471±39.74188min, AUClast分别为46.2976±27.85874min*ng/mL和28.25056±28.34036min*ng/mL,
结论：该方法操作简单,效率高,选择性好,灵敏度高,可成功用于广痛消泡沫气雾剂的健康人体血浆的动力学研究。
Research background and objective
Department of anorectal surgery usually treat diseases mainly include:anal fistula, hemorrhoids, rectum prolapse, rectum and anal canal abscess, rectal cancer, anal fissure, etc. These diseases usually need surgical treatment, But the pain, edema, hemorrhage, infection, delayed wound healing complications often appear after surgical treatments. The postoperative dressing changes become an important means to treat these postoperative complications and promote wound healing in addition to symptomatic treatments.At present, hemorrhoids ointment and hemorrhoids suppository are widely used to treat anorectal diseases, These drugs are supplied through the anus,They play an anti-inflammatory, analgesic, swelling by direct role in the surgical wound or the rectum mucosa. While there are small contact surface at the lesion site, uneven distribution, easy to melt and discharge, short duration of efficacy, low bioavailability. These are based on the above situation, Professor Zhao Baoming developed Guang TongXiao foam aerosol that based on the basic theory of TCM and combined with anorectal disease clinical characteristics,the foam aerosol can not only anti-inflammatory, swelling, relieve spasm and pain but also following advantages:the ejecta no flow, foam stable, long duration in the lesion site, coated with a wide range of uniformly dispersed in the lumen, effectively penetrate the mucosal folds, etc. It's more effective than other formulations,and is a great development potential of dosage form.
GuangTongXiao foam aerosols is a kind of traditional Chinese medicine foam aerosols. Professor Zhao Baoming developed this foam aerosols that is based on the basic theory of TCM and combined with characteristics of anorectal diseases. It is consisted of corydalis, white peony root, rubia cordifolia and othe Chinese medicines, it has spasmolysis analgesia, anti-inflammatory, hemostasis and promote wound healing. Treatment tips say"guang", means small and large intestines;"Tongxiao" means this foam aerosols have analgesia efficacy. Foam aerosol illustrate the formulations of the herbal compound."GuangTongXiao foam aerosols" summed the site of action of the herbal compound, efficacy and dosage forms.The foam aerosol has been finished these researches such as the extraction process、quality standards. efficacy, dosage form advantages and drug security researches by the department of Anorectal Surgery and laboratory of Guang An Men Hospital, the Department of Anorectal Surgery and the central laboratory of Dongzhimen Hospital、 Beijing University of Chinese Medicine the Academy of Military Sciences and Department of Pathology of Peking University Health Science. It has strong research bases because the animal experiments and clinical studies lasted more than20years, but these efficacy reasons unclear, Therefore,this subject will lay the foundation for the future study of the mechanism and guide clinical medication by determination of the active ingredient contents and pharmacokinetic study of healthy volunteers.
Research methods and contents
1.To determine five effective components in the water extract of GuangTongXiao.In this study, using LC-MS/MS method determine simultaneously tetrahydropalmatine, paeoniflorin, berberine, magnolol and honokiol, The results show that The method is rapid and sensitive, attribute strong, This method can determine simultaneously five kinds of effective components. LC-MS/MS method for determining contents of tetrahydropalmatine, paeoniflorin, berberine, magnolol and honokiol, the ibuprofen, raceanisodamine act internal standard. Chromatographic conditions:chromatographic column(Agilent Eclipse Plus C18,100mm×2.1mm,3.5μm), mobile phase A:0.1%of formic acid, B:acetonitrile, gradient elution, the column temperature30℃. Mass spectrometer conditions:ESI ion source, the positive ion mode drying gas temperature300℃, gas flow10L-min-1; Nebulizer pressure30Pa; the capillary voltage positive ions4000V, Paeoniflorin tetrahydropalmatine berberine, magnolol, honokiol, ibuprofen and raceanisodamine monitoring of ion-pair479.2/121,356.2/192.2,336.1/320.2,265.1/247.1,265.2/233.1,205.1/161.1,306.1/140,they are all analysised accurately.Linear ranges:0.8828125-113(r=0.9998);1.54375-49.4(r=0.9995);0.9046875-115.8(r=0.9997);0.88125-112.8(r=0.9994);0.165625-21.2(r=0.9999). spike recovery1.998%,1.14%,3.117%,1.828%and1.596%.The results show that the method high sensitivity, fast analysis,while qualitative ability and quantitative ability good, the method can determine simultaneously five main active ingredient contents in GuangTongXiao aqueous extracts.
2. To study pharmacokinetics of Guangtongxiao foam aerosols that act on healthy human.The health human pharmacokinetics of GuangTongXiao foam aerosols that is administrated by anal.Determining the concentrations of tetrahydropalmatine and berberine in healthy male plasma by LC-MS/MS, raceanisodamine act internal mark, protein of plasma samples was precipitated by acetone,80%acetonitrile extraction, Chromatographic conditions:chromatographic column(Agilent Eclipse Plus C18,100mm×2.1mm,3.5μm), mobile phase A:0.1%of formic acid, B:acetonitrile, gradient elution0～1min(B15%、0.2mL/min);1～2min(B15%～40%,0.3mL/min);2～7min(B40%～80%,0.3mL);7～8min (B80%～15%,0.3mL/min);8～10min(B15%,0.3mL/min);10llmin(B15%,0.2mL/min);12min (B15%、0.2mL/min) Mass spectrometer conditions: ESI ion source, the positive ion mode drying gas temperature300℃, gas flow10L-min-1; Nebulizer pressure30Pa; the capillary voltage positive ions4000V, monitoring of ion-pair are479.2/121,356.2/192.2,306.1/140, the autosampler temperature15℃, the column temperature30℃. methodology evaluation show the tetrahydropalmatine and berberine hydrochloride that in biological samples show a good linear r>0.99, The linear range of the respective components are0.000594-1.188ng/mL,0.000536-1.072ng/mL, Lowest limit of quantification is0.000536ng/ml, intraday precision and day precision are both less than 15%,The extraction recovery are both more than85%. The method is simple, high efficiency, good selectivity, high sensitivity, and can be used successfully in the study of the dynamics of human plasma thay contain GuangTongXiao foam aerosols, On this basis, a high sensitivity, specificity, and accurate LC-MS/MS method has been establed, two kinds of active ingredients in the foam aerosols are determined simultaneously, and research the dynamic characteristics of foam aerosol in healthy male body,in order to provide a reference for the clinical and the late further researches.
引文
[1]傅小勇,等.元胡生物碱的化学研究Ⅵ：元胡叔胺生物碱的反相高效液相色谱法测定[J]·药学学报,1986,21(7)：527.
[2]徐婷,金昔陆,曹惠明.延胡索乙素药理作用的研究进展[J].中国临床药学杂志,2001,10(1)：58-60.
[3]Wai C L, Hui Z, Michael H, etal. Anxiolytic-like action of orally administered dl-tetrahydropalmatine in elevated plusmaze [J], Prog Neuro psychoharmacol Biol Pschiat,2003,27:775-779.
[4]梁健,梁京生,郑平香.延胡索乙素对大鼠局灶性脑缺血再灌注损伤的保护作用[J].中国药理学通报,1998，14(5)：413
[5]梁健,郑平香,王富强,等.延胡索乙素抗脂质过氧化作用与对脑缺血再灌注大鼠行为及病理改变的保护[J].中国药理学通报,1999,15(2)：167
[6]GE X Q, ZHANG H Q, ZHOU H Z, et al. Experimental study oftetrahydrop rotoberberines inhibiting morphine withdrawal syndromes[J]. Chin J D rug Depend,1999,8 (3):182-184.
[7]GE X Q, B IAN C F. Experimental study in treating morphine withdrawal syndromes by a combination of 12tetrahydropalmatine and chlorp romazine and scopolamine [J].//iangsu Clin M ed,2001,5(3):196-198.
[8]朱丽丽,李惠芬.左旋四氢巴马汀对依赖药物作用影响的研究[J].中草药,2005,36(11)：1748-1750
[9]刘嘉,蔡小军,狄留庆.延胡索全碱注射液对麻醉犬血流动力学的影响[J].中国现代药物应用,2008,2(7)：6-8.
[10]刑建峰,王美纳.d1-四氢巴汀对实验性脑血栓形成及体外血小板聚集的抑制作用[J].中国药理学通报,1997,13(3)：258-260.
[11]周向锋,黄斌伦,陈根强.延胡索乙素对大鼠低O2高CO2肺动脉高压及FAK、ERK和Caspase3表达的影响[J].中药药理与临床,2008，24(5)：18-20
[12]俞静,朴炳奎,花宝金.延胡索乙素对放射所致血管内皮细胞损伤的保护作用[J].中华肿瘤防治杂志,2011,18(8)：584-601
[13]邵翎宇,颜洁明,颜梅,等.左旋四氢巴马汀对门静脉压的影响及其机制[J].中国药理学通报,1995,11(3)：248
[14]邵翎宇,颜梅,颜洁明.左旋四氢巴马汀对肝经门脉高压和胰高糖素的影响[J].徐州医学院学报,1995,15(1)：9
[15]唐希灿,金国章,胥彬.延胡索的药理研究Ⅶ.延胡索乙素对狗胃液分泌的影响及催眠作用[J].药学学报,1962,9(3)：145-150.
[16]郝继亭.延胡索的药理研究[N].中国医药导报,2001-12-6(6)
[17]葛晓群,刑淑华,颜梅等.四氢巴马汀对外周儿茶酚胺含量的影响[J].中国药理学通报,1995,11(1)：58-61.
[18]邵翔宇,颜洁明,颜梅.左旋四氛巴马汀对肝经门脉高压和胰高糖素的影响[J].徐州医学院学报,1995,15(1)：9-11.
[19]张国择,谢刚,禹立霞.延胡索总喊对人肝癌细胞系HepG2抑制作用及其对microRNA表达谱的影响[J].南京中医药大学学报,2009,25(3)： 181-183.
[20]崔燎,吴铁.左旋四氢巴马汀增强长春新碱对人白血病细胞株的抑制作用[J].中国药理学通报,1995,11(4)：348
[21]He L, Liu G Q. Interaction of multidrug resistance reversal agents with P-glycoprotein ATPase activity on blood-brain barrier [J].Acta Pharmacol Sin(中国药理学报),2002,23(5)：423-429.
[22]胡定慧,丁建刚,张敏等.延胡索提取物抑菌活性的初步研究[J].辽宁中医杂志,2007,34(4)：496-497.
[23]Zhang Z M, Jiang B, Zheng X X. Effect of 1-tetra-hydropal-matine on expression of adhesion molecules induced by lipopolysaccharides in human umbilical vein endothelium cell[J]. China J Chin Mater Med (中国中药杂志),2005,30(11)：861-864.
[24]徐敬东,杨惠霞,王文,等.延胡索对未孕大鼠离体子宫平滑肌运动的抑制作用[J].中国组织工程研究与临床康复,2007,11(21)：4178-4181
[25]雷汉琴,鞠敏,徐元秀,等.DL-四氢巴马汀和青藤碱对兔输卵管平滑肌活动的影响[J].西安医科大学学报,1993,14(3)：219
[26]闵清,等.延胡索乙素对四氯化碳致小鼠肝损伤的保护作用[J].中国中药杂志,2006,3.31.(6)：483-485
[27]张晓燕,李铣.白芍的化学研究进展[J].沈阳药科大学学报,2002,19(1)：70-73
[28]刘汉珍,刘爱荣,李孝良,等.白芍的化学成分及药理研究进展[J].安徽技术师范学院学报,2001,15(4)：54-57.
[29]李文艳,黄山君,王瑞.中药白芍的药理作用和质量控制研究进展[J].药学服务与研究,2012,12(2)：118-121.
[30]李俊,赵维中,陈敏珠,等.白芍总甙对大鼠腹腔巨噬细胞产生白三烯B4的影响[J].沪国药理学通茂1992,8(1)：36—35
[31]韩连凤,刘发.中药白芍的止痛作用[J].黑龙江医药,2006,19(3)：219
[32]吴慧丽,李慧.白芍总苷对溃疡性结肠炎大鼠细胞因子影响的研究[J].中南药学,2010,8(2)：128-131.
[33]肖尚喜,张咏南,史百芬.白芍总贰促干扰素诱生及抗病毒作用的研究[J].中国药理学通舰1993,9(1)：58-60.
[34]冯文林,伍海涛,罗超华.白芍总苷在消化系统疾病中的药理研究进展[J].时珍国医国药,2012,23(7)：1778-1779
[35]詹可顺,卫华,魏伟.白芍总苷对小鼠化学性肝损伤的保护作用及机制[J].安徽医科大学学报,2006,41(6)：664
[36]李莲.白芍总甙对大鼠酒精性肝损伤保护作用的实验研究[J].湖北职业技术学院学报,2010,13(1)：105.
[37]王华,魏伟,岳莉,等.白芍总苷对卡介苗加脂多糖引起的小鼠免疫性肝损伤的保护作用[J].中国药理学通报,2004,20(8)：875.
[38]周艳丽,张磊,刘维.白芍总苷对雷公藤多苷片所致小鼠急性肝损伤保护作用的实验研究[J].天津中医药,2007,24(1)：61.
[39]魏伟,刘家骏,刘家琴,等.白芍总苷对乙型肝炎的治疗作用及其前景[J].中国药理学通报,2000,16(05)：596.
[40]王永祥,陈敏珠,徐叔云.白芍总苷的镇痛作用[J].中国药理学与毒理学杂志,1988,2(1)：7.
[41]张源潮,孙红胜,潘正论,等.白芍总苷在风湿免疫病中的研究进展[J].世界临床药物,2010,31(8)：449-453.
[42]陈刚,高雪.白芍总苷对巨噬细胞生成PGE2的影响及机制研究[J].中国药理学通报,2011,27(4)：582-583.
[43]傅保娣.芍药苷对缺氧损伤人脐静脉内皮细胞的影响[D].山东：山东大学药学院,2007.
[44]常蕴青,赵中夫,杨柳絮,等.芍药苷对LPS诱导的脐静脉内皮细胞HMGB 1及I CAM-1表达的影响[J].长治医学院学报,2009,23(1)：4-7.
[45]王志强,实伟,刘现兵,等.厚朴体外抑菌作用研究[J].时珍国医国药.2007.18(11)：2763.
[46]王立青,江荣高,陈蕙芳.厚朴酚与和厚朴酚药理作用的研究进展[J].中草药,2005,36(10)：1591-1594.
[47]Xianhe B, Francesca C, Masuko U, et al. Honokiol, a Small Molecular Weight Natural Product, Inhibits Angiogenesis in Vitro and Tumor Growth in Vivo[J]. J Biol Chem,2003,278(37):35501-35507.
[48]Yang SE, Hsieh MT, Tsai TH, et al. Effector mechanism of magnolol induced apoptosis in human lung squamous carcinoma CH27 cells [J]. Br J Pharmacol,2003,138(1):193-201.
[49]Lin SY, Liu JD, Chang HC, et al. Magnolol suppresses proliferation of cultured human colon and liver cancer cells by inhibiting DNA synthesis and activating apotosis[J]. J CellBio-chem,2002,84 (3):532-544
[50]Kenji I, Teru H, Makoto H, et al. Honokiol overcomes conventional drug resistance in human multiple myeloma by induction of case pase-dependent and independent apoptosis [J]. Blood,2005,106:1794-1800.
[51]孙宏伟,李海波.厚朴酚通过自噬途径促进肺癌细胞死亡[J].中医药信息,2010,27(6)：86-89.
[52]李海波,古建军.厚朴酚通过自噬途径促进HepG-2细胞死亡[J].长春中医药大学学报,2010,26(5)：657-659.
[53]杨熙东.厚朴的药理作用及临床应用[J].中国社区医师·医学专业2011,13(287)：151.
[54]李杰萍,梁统,周克元.厚朴酚对大鼠白细胞5-脂氧合酶活性和细胞内钙离子浓度的影响[J].广东医学院学报,2002,20(3)：177-178.
[55]吕江明,陈景,梁剑雄.厚朴干皮“发汗”(加工)前后抗菌镇痛作用的比较研究[J].内蒙古中医药,2004,23(1)：25-26.
[56]陈笈,王伯初.厚朴的药理研究进展[J].重庆大学学报,2005,28(9)：136-139.
[57]陈泉生.厚朴的活性成分厚朴酚(Magnolol)对胃液分泌及实验性溃疡的作用[J].国外医药植物药分册,1981(3)：38.
[58]朱自平,张明发,沈雅琴,等.厚朴对消化系统的药理作用[J].中国中药杂志,1997,11(22)：686-688.
[59]王承南,夏传格.厚朴药理作用及综合利用研究进展[J].经济林研究,2003,21(3)：80-81.
[60]冯瑾,李继遥,周学东.厚朴活性成分对致龋菌生长和产酸影响的体外研究[J].四川大学学报(医学版),2007,38(3)：456-458
[61]Ho K.Y, Tsai CC, Chen CP, et al. Antimicrobial activity of honokiol and magnolol isolated from Magnolia officinalis [J]. Phytother Res,2001,15(2):139-41.
[62]王承南,夏传格.厚朴药理作用及综合利用研究进展[J].经济林研究,2003,21(3)：80-81.
[63]加藤泰基,三木学三.厚朴的药理作用[J].日本东洋医学杂志,1958,3(2)：57.
[64]张启荣,丁立,赵训明.厚朴对兔离体胃肠平滑肌运动的影响[J].陕西医学杂志,2007,36(6)：656-659.
[65]Chiu JH, Ho CT, Wei YH, et al. In vitro and in vivo protec-tive effect of honokiol on rat liver from peroxidative injury [J]. Life Sci,1997,61 (19):1961-1971.
[66]Teng CM, Chen CC, Ko FN, et al. Two antiplatelet agents from Magnolia officinalis [J].Thromb Res,1988,50(6):757-765.
[67]都日娜,乌日娜.黄柏的研究进展[J].中国民族医药杂志.2008,3：3.
[68]赵鲁青,增瑞祥,王森民,等.复方黄柏冷敷剂的药理学研[J].中国药事,1995,9(4)：236.
[69]杨周平,武志军.中药黄柏的药理作用和临床应用研究[J].甘肃医药2010,29(3)：329-331.
[70]南云生,毕晨蕾,等.炮制对黄柏部分药理作用的影响[J].中药材,1995,18(2)：81.
[71]于滢.黄柏的药理作用及应用[J].养殖技术顾问,2011,8：239
[72]胡俊青,胡晓.黄柏化学成分和药理作用的现代研究[J].当代医学,2009,15(7)：139-140.
[73]廖静,鄂征,聂毓秀等.中药黄柏的光敏抗癌作用研究[J].首都医科大学学报,1999,20(3)：153-155.
[74]吴嘉瑞,张冰,张光敏.黄柏药理作用研究进展[J].亚太传统医药,2009,5(11)：160-162
[75]王德金,胡俊英.黄柏胶囊抗炎疗效临床分析[J].中华实用中西医杂志,2004,17(6)：839.
[76]李宗友.黄柏和辽宁木忽木的丁醇提取物刺激PI3激酶和ERK2引起的HepH2细胞中糖原含量的增加[J].国外医学中医中药分册,1999,21(3)：44.
[77]李峰,贾彦竹.黄柏的临床药理作用[J].中医药临床杂志,2004,16(2)：191.
[78]蔡宝昌,潘扬,吴皓,等.国外天然药物抗病毒研究简况[J].国外医学·中药分册,1997,19(3)：48.
[79]宏君.培育捆束水浸应激性溃疡易发系小鼠以及黄柏提取物成分的药理遗传学解析.国外医学-中医中药分册,1992,14(1)：52
[80]张志军.黄柏提取物的抗溃疡效果.国外医学-中医中药分册,1994,16(1)：29
[81]孔令东,杨澄,仇熙,等.黄柏炮制品清除氧自由基和抗脂质过氧化作用[J].中国中药杂志,2001,26(4)：245.
[82]刘斌,杨昭毅,魏伟.近年中药药物动力学研究进展[J].安徽医药,2009,13(10)：1162-1164.
[83]潘海燕.中药药代动力学研究综述[J].实用医技杂志,2008,15(33)：4894- 4896.
[84]高海,李秀岚.动物医学进展,2007,28(8)：94-98.
[85]刘华钢,梁秋云.中药复方药物动力学方法学的国内研究进展[J].2005,8(2)：70-72.
[86]陈慧慧,孙付军,李贵海.中药药代动力学测定方法的研究概况[J].时珍国医国药2010,21(1)：217-219
[87]李淑芳,吴秀君.HPLC法在中药药代动力学中的应用[J].实用药物与临床,2008,11(1)：36.
[88]钟大放.液相色谱-质谱联用法在药物研究中的应用[J].世界科学技术-中医药现代化,2003,5(4)：44.
[89]郭立玮.中药药物动力学方法与应用[M].北京：人民卫生出版社,2002：106,289,297.
[90]伍小波.现代仪器分析方法在中药药动学研究中的应用[J].现代中医药,2004,6：51.
[91]潘嘉,王家葵,邹文俊,等.抑菌效应测定川芎挥发油药动学参数[J].中药药理与临床,2002,18(4)：18.
[92]杨奎,蒲旭峰.论“中药胃肠药动学研究”的意义及对策[J].中国实验实验方剂学杂志,1998,4(1)：36.
[93]赵春丽,刘建芳,刘会臣.中药药物动力学研究进展[J].解放军药学学报2003,19(2)：125-128.
[94]薛燕,雷跻九.中药复方霰弹理论-论中药复方现代研究方法[M].北京：中国环境科学出版杜,1996：4.
[95]颜敏,刘建平.中药药物动力学研究进展[J].药学进展,2005,29(6)：260-265.
[96]苏彦斌,孟宪生,苏彦文,等.中药研究的主体和客体及其关系的分析[J].中华中医药学刊,2007,25(2)：342-343.
[97]何绍雄,宋开源,苏兆虞,等.时间药理学与时间治疗学[M].天津：天津科学技术出版社,1994：261-267.
[98]张家玮,孙建宁,张爱林.有效成分组-中药复方活性物质基础研究方法刍议[J].中国中医药信息杂志,2006,13(2)：2-5.
[2]徐婷,金昔陆,曹惠明.延胡索乙素药理作用的研究进展[J].中国临床药学杂志,2001,10(1)：58-60.
[3]Wai C L, Hui Z, Michael H, etal. Anxiolytic-like action of orally administered dl-tetrahydropalmatine in elevated plusmaze [J], Prog Neuro psychoharmacol Biol Pschiat,2003,27:775-779.
[4]梁健,梁京生,郑平香.延胡索乙素对大鼠局灶性脑缺血再灌注损伤的保护作用[J].中国药理学通报,1998，14(5)：413
[5]梁健,郑平香,王富强,等.延胡索乙素抗脂质过氧化作用与对脑缺血再灌注大鼠行为及病理改变的保护[J].中国药理学通报,1999,15(2)：167
[6]GE X Q, ZHANG H Q, ZHOU H Z, et al. Experimental study oftetrahydrop rotoberberines inhibiting morphine withdrawal syndromes[J]. Chin J D rug Depend,1999,8 (3):182-184.
[7]GE X Q, B IAN C F. Experimental study in treating morphine withdrawal syndromes by a combination of 12tetrahydropalmatine and chlorp romazine and scopolamine [J].//iangsu Clin M ed,2001,5(3):196-198.
[8]朱丽丽,李惠芬.左旋四氢巴马汀对依赖药物作用影响的研究[J].中草药,2005,36(11)：1748-1750
[9]刘嘉,蔡小军,狄留庆.延胡索全碱注射液对麻醉犬血流动力学的影响[J].中国现代药物应用,2008,2(7)：6-8.
[10]刑建峰,王美纳.d1-四氢巴汀对实验性脑血栓形成及体外血小板聚集的抑制作用[J].中国药理学通报,1997,13(3)：258-260.
[11]周向锋,黄斌伦,陈根强.延胡索乙素对大鼠低O2高CO2肺动脉高压及FAK、ERK和Caspase3表达的影响[J].中药药理与临床,2008，24(5)：18-20
[12]俞静,朴炳奎,花宝金.延胡索乙素对放射所致血管内皮细胞损伤的保护作用[J].中华肿瘤防治杂志,2011,18(8)：584-601
[13]邵翎宇,颜洁明,颜梅,等.左旋四氢巴马汀对门静脉压的影响及其机制[J].中国药理学通报,1995,11(3)：248
[14]邵翎宇,颜梅,颜洁明.左旋四氢巴马汀对肝经门脉高压和胰高糖素的影响[J].徐州医学院学报,1995,15(1)：9
[15]唐希灿,金国章,胥彬.延胡索的药理研究Ⅶ.延胡索乙素对狗胃液分泌的影响及催眠作用[J].药学学报,1962,9(3)：145-150.
[16]郝继亭.延胡索的药理研究[N].中国医药导报,2001-12-6(6)
[17]葛晓群,刑淑华,颜梅等.四氢巴马汀对外周儿茶酚胺含量的影响[J].中国药理学通报,1995,11(1)：58-61.
[18]邵翔宇,颜洁明,颜梅.左旋四氛巴马汀对肝经门脉高压和胰高糖素的影响[J].徐州医学院学报,1995,15(1)：9-11.
[19]张国择,谢刚,禹立霞.延胡索总喊对人肝癌细胞系HepG2抑制作用及其对microRNA表达谱的影响[J].南京中医药大学学报,2009,25(3)： 181-183.
[20]崔燎,吴铁.左旋四氢巴马汀增强长春新碱对人白血病细胞株的抑制作用[J].中国药理学通报,1995,11(4)：348
[21]He L, Liu G Q. Interaction of multidrug resistance reversal agents with P-glycoprotein ATPase activity on blood-brain barrier [J].Acta Pharmacol Sin(中国药理学报),2002,23(5)：423-429.
[22]胡定慧,丁建刚,张敏等.延胡索提取物抑菌活性的初步研究[J].辽宁中医杂志,2007,34(4)：496-497.
[23]Zhang Z M, Jiang B, Zheng X X. Effect of 1-tetra-hydropal-matine on expression of adhesion molecules induced by lipopolysaccharides in human umbilical vein endothelium cell[J]. China J Chin Mater Med (中国中药杂志),2005,30(11)：861-864.
[24]徐敬东,杨惠霞,王文,等.延胡索对未孕大鼠离体子宫平滑肌运动的抑制作用[J].中国组织工程研究与临床康复,2007,11(21)：4178-4181
[25]雷汉琴,鞠敏,徐元秀,等.DL-四氢巴马汀和青藤碱对兔输卵管平滑肌活动的影响[J].西安医科大学学报,1993,14(3)：219
[26]闵清,等.延胡索乙素对四氯化碳致小鼠肝损伤的保护作用[J].中国中药杂志,2006,3.31.(6)：483-485
[27]张晓燕,李铣.白芍的化学研究进展[J].沈阳药科大学学报,2002,19(1)：70-73
[28]刘汉珍,刘爱荣,李孝良,等.白芍的化学成分及药理研究进展[J].安徽技术师范学院学报,2001,15(4)：54-57.
[29]李文艳,黄山君,王瑞.中药白芍的药理作用和质量控制研究进展[J].药学服务与研究,2012,12(2)：118-121.
[30]李俊,赵维中,陈敏珠,等.白芍总甙对大鼠腹腔巨噬细胞产生白三烯B4的影响[J].沪国药理学通茂1992,8(1)：36—35
[31]韩连凤,刘发.中药白芍的止痛作用[J].黑龙江医药,2006,19(3)：219
[32]吴慧丽,李慧.白芍总苷对溃疡性结肠炎大鼠细胞因子影响的研究[J].中南药学,2010,8(2)：128-131.
[33]肖尚喜,张咏南,史百芬.白芍总贰促干扰素诱生及抗病毒作用的研究[J].中国药理学通舰1993,9(1)：58-60.
[34]冯文林,伍海涛,罗超华.白芍总苷在消化系统疾病中的药理研究进展[J].时珍国医国药,2012,23(7)：1778-1779
[35]詹可顺,卫华,魏伟.白芍总苷对小鼠化学性肝损伤的保护作用及机制[J].安徽医科大学学报,2006,41(6)：664
[36]李莲.白芍总甙对大鼠酒精性肝损伤保护作用的实验研究[J].湖北职业技术学院学报,2010,13(1)：105.
[37]王华,魏伟,岳莉,等.白芍总苷对卡介苗加脂多糖引起的小鼠免疫性肝损伤的保护作用[J].中国药理学通报,2004,20(8)：875.
[38]周艳丽,张磊,刘维.白芍总苷对雷公藤多苷片所致小鼠急性肝损伤保护作用的实验研究[J].天津中医药,2007,24(1)：61.
[39]魏伟,刘家骏,刘家琴,等.白芍总苷对乙型肝炎的治疗作用及其前景[J].中国药理学通报,2000,16(05)：596.
[40]王永祥,陈敏珠,徐叔云.白芍总苷的镇痛作用[J].中国药理学与毒理学杂志,1988,2(1)：7.
[41]张源潮,孙红胜,潘正论,等.白芍总苷在风湿免疫病中的研究进展[J].世界临床药物,2010,31(8)：449-453.
[42]陈刚,高雪.白芍总苷对巨噬细胞生成PGE2的影响及机制研究[J].中国药理学通报,2011,27(4)：582-583.
[43]傅保娣.芍药苷对缺氧损伤人脐静脉内皮细胞的影响[D].山东：山东大学药学院,2007.
[44]常蕴青,赵中夫,杨柳絮,等.芍药苷对LPS诱导的脐静脉内皮细胞HMGB 1及I CAM-1表达的影响[J].长治医学院学报,2009,23(1)：4-7.
[45]王志强,实伟,刘现兵,等.厚朴体外抑菌作用研究[J].时珍国医国药.2007.18(11)：2763.
[46]王立青,江荣高,陈蕙芳.厚朴酚与和厚朴酚药理作用的研究进展[J].中草药,2005,36(10)：1591-1594.
[47]Xianhe B, Francesca C, Masuko U, et al. Honokiol, a Small Molecular Weight Natural Product, Inhibits Angiogenesis in Vitro and Tumor Growth in Vivo[J]. J Biol Chem,2003,278(37):35501-35507.
[48]Yang SE, Hsieh MT, Tsai TH, et al. Effector mechanism of magnolol induced apoptosis in human lung squamous carcinoma CH27 cells [J]. Br J Pharmacol,2003,138(1):193-201.
[49]Lin SY, Liu JD, Chang HC, et al. Magnolol suppresses proliferation of cultured human colon and liver cancer cells by inhibiting DNA synthesis and activating apotosis[J]. J CellBio-chem,2002,84 (3):532-544
[50]Kenji I, Teru H, Makoto H, et al. Honokiol overcomes conventional drug resistance in human multiple myeloma by induction of case pase-dependent and independent apoptosis [J]. Blood,2005,106:1794-1800.
[51]孙宏伟,李海波.厚朴酚通过自噬途径促进肺癌细胞死亡[J].中医药信息,2010,27(6)：86-89.
[52]李海波,古建军.厚朴酚通过自噬途径促进HepG-2细胞死亡[J].长春中医药大学学报,2010,26(5)：657-659.
[53]杨熙东.厚朴的药理作用及临床应用[J].中国社区医师·医学专业2011,13(287)：151.
[54]李杰萍,梁统,周克元.厚朴酚对大鼠白细胞5-脂氧合酶活性和细胞内钙离子浓度的影响[J].广东医学院学报,2002,20(3)：177-178.
[55]吕江明,陈景,梁剑雄.厚朴干皮“发汗”(加工)前后抗菌镇痛作用的比较研究[J].内蒙古中医药,2004,23(1)：25-26.
[56]陈笈,王伯初.厚朴的药理研究进展[J].重庆大学学报,2005,28(9)：136-139.
[57]陈泉生.厚朴的活性成分厚朴酚(Magnolol)对胃液分泌及实验性溃疡的作用[J].国外医药植物药分册,1981(3)：38.
[58]朱自平,张明发,沈雅琴,等.厚朴对消化系统的药理作用[J].中国中药杂志,1997,11(22)：686-688.
[59]王承南,夏传格.厚朴药理作用及综合利用研究进展[J].经济林研究,2003,21(3)：80-81.
[60]冯瑾,李继遥,周学东.厚朴活性成分对致龋菌生长和产酸影响的体外研究[J].四川大学学报(医学版),2007,38(3)：456-458
[61]Ho K.Y, Tsai CC, Chen CP, et al. Antimicrobial activity of honokiol and magnolol isolated from Magnolia officinalis [J]. Phytother Res,2001,15(2):139-41.
[62]王承南,夏传格.厚朴药理作用及综合利用研究进展[J].经济林研究,2003,21(3)：80-81.
[63]加藤泰基,三木学三.厚朴的药理作用[J].日本东洋医学杂志,1958,3(2)：57.
[64]张启荣,丁立,赵训明.厚朴对兔离体胃肠平滑肌运动的影响[J].陕西医学杂志,2007,36(6)：656-659.
[65]Chiu JH, Ho CT, Wei YH, et al. In vitro and in vivo protec-tive effect of honokiol on rat liver from peroxidative injury [J]. Life Sci,1997,61 (19):1961-1971.
[66]Teng CM, Chen CC, Ko FN, et al. Two antiplatelet agents from Magnolia officinalis [J].Thromb Res,1988,50(6):757-765.
[67]都日娜,乌日娜.黄柏的研究进展[J].中国民族医药杂志.2008,3：3.
[68]赵鲁青,增瑞祥,王森民,等.复方黄柏冷敷剂的药理学研[J].中国药事,1995,9(4)：236.
[69]杨周平,武志军.中药黄柏的药理作用和临床应用研究[J].甘肃医药2010,29(3)：329-331.
[70]南云生,毕晨蕾,等.炮制对黄柏部分药理作用的影响[J].中药材,1995,18(2)：81.
[71]于滢.黄柏的药理作用及应用[J].养殖技术顾问,2011,8：239
[72]胡俊青,胡晓.黄柏化学成分和药理作用的现代研究[J].当代医学,2009,15(7)：139-140.
[73]廖静,鄂征,聂毓秀等.中药黄柏的光敏抗癌作用研究[J].首都医科大学学报,1999,20(3)：153-155.
[74]吴嘉瑞,张冰,张光敏.黄柏药理作用研究进展[J].亚太传统医药,2009,5(11)：160-162
[75]王德金,胡俊英.黄柏胶囊抗炎疗效临床分析[J].中华实用中西医杂志,2004,17(6)：839.
[76]李宗友.黄柏和辽宁木忽木的丁醇提取物刺激PI3激酶和ERK2引起的HepH2细胞中糖原含量的增加[J].国外医学中医中药分册,1999,21(3)：44.
[77]李峰,贾彦竹.黄柏的临床药理作用[J].中医药临床杂志,2004,16(2)：191.
[78]蔡宝昌,潘扬,吴皓,等.国外天然药物抗病毒研究简况[J].国外医学·中药分册,1997,19(3)：48.
[79]宏君.培育捆束水浸应激性溃疡易发系小鼠以及黄柏提取物成分的药理遗传学解析.国外医学-中医中药分册,1992,14(1)：52
[80]张志军.黄柏提取物的抗溃疡效果.国外医学-中医中药分册,1994,16(1)：29
[81]孔令东,杨澄,仇熙,等.黄柏炮制品清除氧自由基和抗脂质过氧化作用[J].中国中药杂志,2001,26(4)：245.
[82]刘斌,杨昭毅,魏伟.近年中药药物动力学研究进展[J].安徽医药,2009,13(10)：1162-1164.
[83]潘海燕.中药药代动力学研究综述[J].实用医技杂志,2008,15(33)：4894- 4896.
[84]高海,李秀岚.动物医学进展,2007,28(8)：94-98.
[85]刘华钢,梁秋云.中药复方药物动力学方法学的国内研究进展[J].2005,8(2)：70-72.
[86]陈慧慧,孙付军,李贵海.中药药代动力学测定方法的研究概况[J].时珍国医国药2010,21(1)：217-219
[87]李淑芳,吴秀君.HPLC法在中药药代动力学中的应用[J].实用药物与临床,2008,11(1)：36.
[88]钟大放.液相色谱-质谱联用法在药物研究中的应用[J].世界科学技术-中医药现代化,2003,5(4)：44.
[89]郭立玮.中药药物动力学方法与应用[M].北京：人民卫生出版社,2002：106,289,297.
[90]伍小波.现代仪器分析方法在中药药动学研究中的应用[J].现代中医药,2004,6：51.
[91]潘嘉,王家葵,邹文俊,等.抑菌效应测定川芎挥发油药动学参数[J].中药药理与临床,2002,18(4)：18.
[92]杨奎,蒲旭峰.论“中药胃肠药动学研究”的意义及对策[J].中国实验实验方剂学杂志,1998,4(1)：36.
[93]赵春丽,刘建芳,刘会臣.中药药物动力学研究进展[J].解放军药学学报2003,19(2)：125-128.
[94]薛燕,雷跻九.中药复方霰弹理论-论中药复方现代研究方法[M].北京：中国环境科学出版杜,1996：4.
[95]颜敏,刘建平.中药药物动力学研究进展[J].药学进展,2005,29(6)：260-265.
[96]苏彦斌,孟宪生,苏彦文,等.中药研究的主体和客体及其关系的分析[J].中华中医药学刊,2007,25(2)：342-343.
[97]何绍雄,宋开源,苏兆虞,等.时间药理学与时间治疗学[M].天津：天津科学技术出版社,1994：261-267.
[98]张家玮,孙建宁,张爱林.有效成分组-中药复方活性物质基础研究方法刍议[J].中国中医药信息杂志,2006,13(2)：2-5.
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