吴茱萸不同炮制品中吴茱萸碱、吴茱萸次碱、柠檬苦素含量测定

目的:考察不同炮制方法及炮制前后吴茱萸中吴茱萸碱、吴茱萸次碱和柠檬苦素的含量变化情况。方法:采用高效液相色谱法,Hypersil ODS C₁₈（250 mm×4.6 mm,5μm）;流动相:乙腈-水-四氢呋喃-乙酸（41:59:1:0.2）;流速:1.0 ml·min^-1;检测波长:225 nm;柱温:25℃。结果:吴茱萸碱、吴茱萸次碱、柠檬苦素进样量分别在0.156～3.120μg（r=0.999 6,n=6）、0.116～2.320μg（r=1.000 0,n=6）、0.620～12.400μg（r=0.999 1,n=6）范围内与峰面积呈良好线性关系,平均回收率分别为97.2%（RSD=1.33%）、97.1%（RSD=1.56%）、97.3%（RSD=1.52%）。结论:不同炮制方法吴茱萸碱、吴茱萸次碱、柠檬苦素含量不同,砂烫法优于其他炮制方法。     

HPLC方法测定华佗再造丸中阿魏酸、吴茱萸内酯、吴茱萸碱和吴茱萸内碱的研究

目的 建立HPLC法测定华佗再造丸中阿魏酸、吴茱萸内酯、吴茱萸碱及吴茱萸次碱的含量.方法 采用Waters SunFire C18（4.6mm×250mm,5μm）色谱柱,以乙腈-0.1%磷酸进行梯度洗脱,流速1.0mL·min-1,阿魏酸检测波长为323nm,吴茱萸内酯、吴茱萸碱检测波长为225nm,吴茱萸次碱检测波长为343nm,柱温30℃.结果 表明阿魏酸、吴茱萸内酯、吴茱萸碱和吴茱萸次碱分别在3.91～39.10μg·mL-1（r=1.0000）、10.70～64.20μg·mL-1（r=0.9998）、7.60～76.00μg·mL-1（r=0.9999）、3.65～36.50μg·mL-1（r=0.9999）范围内与峰面积呈良好的线性关系;平均回收率（n=6）分别为96.6%（RSD=1.0%）、98.1%（RSD=2.2%）、100.3%（RSD=0.8%）、95.3%（RSD=1.2%）.结论 本文方法简便准确,可用于华佗再造丸中阿魏酸、吴茱萸内酯、吴茱萸碱及吴茱萸次碱的含量测定.     

HPLC法同时测定吴茱萸中吴茱萸碱、吴茱萸次碱与吴茱萸内酯的含量

目的:建立同时测定吴茱萸中吴茱萸碱、吴茱萸次碱与吴茱萸内酯含量的方法。方法:采用高效液相色谱法。色谱柱为迪马C1(8200mm×4.6mm,5μm),流动相为乙腈-0.04%庚烷磺酸钠溶液(48:52,V/V),流速为1.0mL·min-1,检测波长为225nm,柱温为35℃。结果:吴茱萸碱、吴茱萸次碱、吴茱萸内酯的进样浓度分别在5.38～53.80、5.02～50.20、10.30～103.00μg·mL-(1r均为0.9999)范围内与各自峰面积积分值呈良好线性关系;三者平均加样回收率分别为99.75%、97.66%、85.68%,RSD分别为0.67%、1.16%、1.54%(n=6)。结论:本方法简便、可行、重复性好,可用于吴茱萸中吴茱萸碱和吴茱萸次碱的含量测定;但吴茱萸内酯的回收率较低,需进一步改进其测定方法。     

HPLC法测定超微戊己丸中吴茱萸碱和吴茱萸次碱含量

目的：建立超微戊己丸中吴茱萸碱和吴茱萸次碱的含量测定方法。方法采用Kromasail C18色谱柱（250mm×4.6mm,5μm）,流动相为流速1.0mL/min的乙腈-乙腈10%（50：50）,测定波长为225nm。结果吴茱萸碱和吴茱萸次碱理论板数分别为2681和2067,Y吴茱萸碱=1.965×10^-7X ＋0.07646,r=0.9999）, Y吴茱萸次碱=3.658×10^-7X-0.2199,r=0.9999）,吴茱萸碱与吴茱萸次碱的线性区间分别为10.2～51.0μg/mL以及10.0～50.0μg/mL ,两者平均回收率分别为97.3%和101.4%,其相对标准偏差分别为3.2%和3.9%,最低检测限为0.05μg/mL和0.1μg/mL。结论使用高效液相色谱法对超微戊己丸当中的吴茱萸碱以及吴茱萸次碱含量进行检测的精确率较高,重现性好。     

不同生产工艺的戊己丸中吴茱萸碱和吴茱萸次碱含质量比较

目的:建立戊己丸中吴茱萸碱和吴茱萸次碱含质量的高效液相(HPLC)测定方法,并对超微戊己丸和常规戊己丸中吴茱萸碱和吴茱萸次碱的含质量进行比较。方法:采用色谱柱HypersilC18柱(4.6mm×250mm,5μm),流动相:乙腈水(48∶52),流速:1mL/min,检测波长:225nm。结果:吴茱萸碱和吴茱萸次碱的理论塔板数均大于3000,吴茱萸碱回归方程:A吴茱萸碱=107.73ρ-14.331,r=0.9999,线性范围1.02～20.4μg/mL;吴茱萸次碱回归方程:A吴茱萸次碱=15.596ρ-45.09,r=0.9998,线性范围4.4～176μg/mL。超微戊己丸中吴茱萸碱平均回收率为:99.61%,RSD为0.50%,质量分数为(0.096±0.0148)%;吴茱萸次碱平均回收率为94.85%,RSD为1.00%,质量分数为(0.343±0.0467)%。常规戊己丸中吴茱萸碱平均回收率为98.56%,RSD为1.70%,质量分数为(0.81±0.003)%;吴茱萸次碱平均回收率为99.55%,RSD为0.50%,质量分数为(0.161±0.0075)%。结论:超微戊己丸中吴茱萸碱和吴茱萸次碱的质量分数高于常规戊己丸。     

HPLC同时测定肠康片中吴茱萸碱、吴茱萸次碱、木香烃内酯和去氢木香内酯的含量

目的 建立肠康片中吴茱萸碱、吴茱萸次碱、木香烃内酯和去氢木香内酯的HPLC含量测定方法。方法 色谱柱为岛津Eclipse-XDB-C₁₈柱(250 mm×4.6 mm, 5μm),流动相为乙腈-0.02%磷酸(40∶60),流速1.0 mL·min^-1,柱温为35℃,检测波长为225 nm。结果 吴茱萸碱y=102097.5127x+1168.9695(r=0.9999),线性范围为0.5016～5.0160μg·mL^-1,平均回收率为101.52%,RSD=1.87%(n=6);吴茱萸次碱：y=524410184x-316.7073(r=0.9998),线性范围为0.5021～5.0209μg·mL^-1,平均回收率为101.56%、RSD=2.06%(n=6);木香烃内酯：y=1757575.24234x-75774.20833(r=0.9996),线性范围为0.0299～0.2993 mg·mL^-1,平均回收率为100.05%、RSD=1.26%(n=6);去氢木香内酯：y...     

超临界流体萃取吴茱萸中吴茱萸碱和吴茱萸次碱

目的研究超临界流体萃取(SFE)吴茱萸中吴茱萸碱和吴茱萸次碱的提取工艺,并用高效液相色谱法(HPLC)测定吴茱萸碱和吴茱萸次碱的含量.方法采用95%乙醇作夹带剂,运用超临界流体的萃取工艺,含量测定时,在Water-C18柱上,以乙腈-水(43∶57)为流动相,流速1.0 mL*min-1,检测波长290 nm,柱温40℃.结果吴茱萸碱和吴茱萸次碱的标准曲线在0.024～0.120 μg(r=0.9994)和0.026～0.130 μg(r=0.9995)之间有较好的线性关系;加样平均回收率分别为102.7%(RSD=1.58)和101.7%(RSD=1.79%). 结论采用SFE萃取吴茱萸中吴茱萸碱和吴茱萸次碱的提取工艺速度快,HPLC法测定吴茱萸碱和吴茱萸次碱的含量精确,重现性好.     

HPLC法同时测定不同配伍比例黄连-吴茱萸药对中4种有效成分的含量

目的:建立同时测定黄连-吴茱萸药对中4种有效成分含量的方法,探讨该药对的合理比例。方法:采用高效液相色谱法。色谱柱为Welch XB C_(18),流动相为乙睛-1.5 mmol/L十二烷基硫酸钠溶液(磷酸调p H至5.0)(梯度洗脱),流速为1.0 mL/min,检测波长为265 nm,柱温为30℃,进样量为10μL。结果:盐酸小檗碱、盐酸巴马汀、吴茱萸碱、吴茱萸次碱检测质量浓度线性范围分别为2.24～44.80μg/m L(r=0.999 9)、1.26～31.50μg/mL(r=0.999 8)、2.70～81.00μg/mL(r=0.999 8)、1.65～49.50μg/mL(r=0.999 9);精密度、稳定性、重复性试验的RSD<3.0%;加样回收率分别为98.11%～100.73%(RSD=1.04%,n=6)、96.54%～103.47%(RSD=1.86%,n=6)、95.49%～102.36%(RSD=2.05%,n=6)、97.19%～103.24%(RSD=2.19%,n=6)。当黄连-吴茱萸比例(m/m)为2∶1时,各成分综合含量最高。结论:该方法操作简便,精密度、稳定性、重复性好,可用于不同配伍比例黄连-吴茱萸药对中4种有效成分含量的同时测定;该药对中黄连-吴茱萸比例(m/m)为2∶1较合理。     

反相高效液相色谱法同时测定复方木香小檗碱片中吴茱萸碱和吴茱萸次碱的含量

目的建立复方木香小檗碱片中吴茱萸碱与吴茱萸次碱含量测定的方法。方法采用RP-HPLC法,ODS2柱（250mm×4．6mm,5μm）,以乙腈-水（39：61）为流动相;流速为1.0mL·min^-1;检测波长为225nm;柱温为35℃;进样量为20μL。结果吴菜萸碱在2．096～10．480μg·mL^-1与峰面积有良好的线性关系,其平均回收率为98．76％,方法精密度（RSD）为1．68％（n=6）;吴茱萸次碱在2．816～14．080μg·mL^-1与峰面积有艮好的线性关系,其平均回收率为99．54％,方法精密度（RSD）为1．93％（n=6）。结论所建立的方法简便易行、准确可靠,可用于复方木香小檗碱片中吴茱萸碱与吴茱萸次碱的含量测定。     

高效液相色谱法测定小儿腹泻凝胶膏中吴茱萸碱及吴茱萸次碱含量

目的建立测定小儿腹泻凝胶膏中吴茱萸碱及吴茱萸次碱含量的高效液相色谱(HPLC)法。方法小儿腹泻凝胶膏中加入80%乙醇超声提取吴茱萸碱及吴茱萸次碱,采用HPLC法,色谱柱为Cosmosil 5 C18-MS-Ⅱ柱(250 mm×4.6 mm,5μm),乙腈-水(51∶49)为流动相,流速为1.0 mL/min,检测波长为225 nm,进样量为10μL。结果吴茱萸碱和吴茱萸次碱进样量分别在0.089～0.89μg和0.077～0.768μg范围内与峰面积呈良好线性关系,吴茱萸碱和吴茱萸次碱的平均回收率为97.80%和99.87%,RSD为0.88%和1.27%。结论该方法简单、准确、重现性好,可用于小儿腹泻凝胶膏中吴茱萸碱及吴茱萸次碱的含量测定。     

吴茱萸次碱在犬体内的药物动力学规律

目的 研究吴茱萸次碱在犬体内的药动学规律.方法 采用三交叉实验设计,6条Beagle犬分别单剂量静脉注射吴茱萸次碱,在预先设定的时间点采集血样,血浆中吴茱萸次碱浓度用HPLC检测, 浓度-时间数据用DAS (V. 2.0)药动学软件分析,计算药动学参数,分析AUC及ke 与剂量的线性关系.结果 给予犬静脉注射0. 25、0.42、0.58 mg·kg-1 吴茱萸次碱后的t1/2 分别为11.969、14.165、14.206 min;AUC0 -∞分别为3.715、9.999、15.483 mg·min·L-1;ke分别为0.048、0.044、0.049.各剂量组的t1/2及ke 与剂量无关(P＞0. 05) .结论 静脉给药后,吴茱萸次碱在犬体内的动力学过程符合一室模型,在实验所用剂量范围内为一级线性动力学过程.     

Induction of cytochrome P450s by rutaecarpine and metabolism of rutaecarpine by cytochrome P450s

Rutaecarpine is an alkaloid originally isolated from the unripe fruit of Evodia rutaecarpa. Recently, rutaecarpine has been characterized to have an anti-inflammatory activity through cyclooxygenase-2 inhibition. In the present studies, the effects of rutaecarpine on liver cytochrome P450 s (P450s) and P450 s involved in the metabolism of rutaecarpine were studied in vivo and in vitro, respectively, because the data are crucial in the early development of rutaecarpine as a new drug candidate. Oral administration to male ICR mice of rutaecarpine for 3 consecutive days induced liver P450 1A-, 2B- and 2E1-selective monooxygenase activities. The induction of P450 1A and 2B by rutaecarpine was confirmed by Western immunoblotting. When rutaecarpine was incubated with rat liver microsomes in the presence of an NADPH-generating system, five metabolites were detected by UV and mass spectral analyses. The 3-methylcholanthrene- and phenobarbital-induced microsomes greatly increased the formation of metabolites. Our present results suggest that rutaecarpine might induce P450 1A and 2B in mice, and that P450 1A and 2B might predominantly metabolize rutaecarpine in rat liver microsomes.     

吴茱萸次碱对心脏过敏损伤的保护作用

目的 观察吴茱萸次碱对心脏过敏损伤的保护作用与激活辣椒素受体。剌激内源性降钙素基因相关肽(CGRP)释放的关系。方法 预致敏的离体心脏用K-H液恒压逆行灌流,从侧管中注入牛血清白蛋白(5 mg),造成过敏损伤。测定冠脉流出液中CGRP和心肌组织中肿瘤坏死因子(TNF-α)的浓度。结果预致敏的豚鼠心脏受抗原(牛血清蛋白)攻击引起冠脉流量(CF)显著减少,左室内压(LVP)和左室内压最大变化速率(±dp/dtmax)降低,同时伴有心率(HR)的增加和P-R间期的延长。两个浓度吴莱萸次碱(0.3或1.0 μmol·L-1)均能显著抑制抗原攻击所致的心功能抑制,表现为LVP、±dp/dtmax和CF升高,P-R间期明显缩短,并能同时促进CGRP的释放和降低心肌组织TNF-α的浓度。昊茱萸次碱对窦性心动过速无明显影响。吴茱萸次碱对心脏过敏损伤的保护作用能被一种选择性CGRP受体拮抗剂CGRP8-37所取消。结论 吴茱萸次碱对心脏过敏损伤的保护作用是通过促进CGRP释放所介导,其心脏保护作用可能与抑制心肌组织TNF-α产生有关。     

吴茱萸次碱对小鼠溃疡性肠炎的治疗作用

目的:观察吴茱萸次碱治疗小鼠急性结肠炎的药效及对豚鼠肠道平滑肌运动的影响。方法:2,4-二硝基氯苯(DNCB)／乙醇灌肠制作小鼠急性结肠炎模型,经口给予吴茱萸次碱10、30、100 mg·kg-1,观察吴茱萸次碱对小鼠腹泻,组织形态损伤,结肠组织PGE2含量、MPO活力以及对乙酰胆碱、组胺作用下豚鼠离体肠管收缩运动的影响。结果:DNCB诱发结肠炎后,小鼠出现腹泻、充血、溃疡、肠壁增厚等变化。经口给予吴茱萸次碱后,可以显著改善小鼠的腹泻、组织损伤,并且能够降低结肠组织中MPO活性和PGE2的含量,对乙酰胆碱及组胺作用下豚鼠离体肠管收缩亦表现出显著的对抗作用。结论:吴茱萸次碱可以抑制炎性浸润、渗出和组织增生,减轻结肠黏膜的病理损害和抑制肠道平滑肌运动,对实验性动物急性结肠炎有较好的治疗作用。     

HPLC测定左金丸中吴茱萸次碱含量

目的:建立高效液相色谱法测定左金丸中吴茱萸次碱含量的方法。方法:采用高效液相色谱法。IntersilC_(18)分析色谱柱(4.6mm ID×250mm,粒径5μm),流动相为乙腈—10％乙腈(50:50),流速1ml·min~(-1),检测波长345nm。结果:吴茱萸次碱的理论板数为2100。回归方程Y=-0.2197+0.0000003484X(r=0.9999),线性范围12～60μm·ml~(-1)。平均回收率为96.9％(n=5),RSD 3.7％,最低检出浓度为0.lμg·ml~(-1)。结论:方法简便,结果准确。     

吴茱萸次碱对高血压大鼠血压的影响及作用机制

目的观察吴茱萸次碱对高血压模型大鼠血压的影响,并初步探讨其作用机制。方法分别采用自发性高血压大鼠(SHR)及两肾一夹(2K1C)高血压大鼠模型,设正常对照组、模型组、卡托普利阳性组及吴茱萸次碱低、中、高剂量组(10,20,40 mg/kg),每日灌胃给药1次,给药4周,每周测定1次血压,末次给药后,腹主动脉取血,分别测定血浆血栓素(TXB2)、6-酮-前列腺-F1α(6-Keto-PGF1α)、肾素活性(PRA)、心房钠肽(ANP)水平。结果在两个模型中吴茱萸次碱均有较好的降压作用,并能使SHR大鼠血浆TXB2水平降低,6-Keto-PGF1α水平升高;使2K1C大鼠血浆PRA、ANP水平升高。结论吴茱萸次碱能明显降低SHR及2K1C大鼠血压,其降压效果可能通过调节PGI2和TXA2水平,改善血管内皮功能及增加舒血管物质ANP水平实现的。     

吴茱萸次碱的研究进展

吴茱萸次碱是中药吴茱萸中的主要有效成分,本文介绍了近十余年来国内外对吴茱萸次碱的研究进展,包括合成、对药物代谢酶的影响、代谢机制和药理作用等。     

吴茱萸碱和吴茱萸次碱的药理学研究进展

介绍吴茱萸碱和吴茱萸次碱的药理学研究进展 ,包括 :心血管作用、抗血小板聚集和血栓形成、抗癌、抗炎、镇痛、对内分泌系统的作用、减肥和体温调节作用、对非血管平滑肌的作用、抗缺氧作用等     

Mechanism-based inhibition of CYPs and RMs-induced hepatoxicity by rutaecarpine

Abstract

1.?Rutaecarpine, a quinolone alkaloid isolated from the unripe fruit of Evodia rutaecarpa, is one of the main active components used in a variety of clinical applications, including the treatment of hypertension and arrhythmia. However, its hepatotoxicity has also been reported in recent years.

2.?Reactive metabolites (RMs) play a vital role in drug-induced liver injury. Rutaecarpine has a secondary amine structure that may be activated to RMs. The aim of the study was to investigate the inhibition of rutaecarpine on CYPs and explore the possible relationship between RMs and potential hepatotoxicity.

3.?A cell counting kit-8 cytotoxicity assay indicated that rutaecarpine can decrease the primary rat hepatocyte viability, increase lactate dehydrogenase and reactive oxygen species, reduce JC-1, and cause cell stress and membrane damage. The indexes were significantly restored by adding ABT, an inhibitor of CYPs. A cocktail assay showed that CYP1A2, CYP2C9, CYP2C19, CYP2E1 and CYP3A4 can be inhibited by rutaecarpine in human liver microsomes. The IC₅₀ values of CYP1A2 with and without NADPH were 2.2 and 7.4?μM, respectively, which presented a 3.3 shift. The results from a metabolic assay indicated that three mono-hydroxylated metabolites and two di-hydroxylated metabolites were identified and two GSH conjugates were also trapped.

4.?Rutaecarpine can inhibit the activities of CYPs and exhibit a potential mechanism-based inhibition on CYP1A2. RMs may cause herb–drug interactions, providing important information for predicting drug-induced hepatotoxicity.     

Immunosuppressive effects of rutaecarpine in female BALB/c mice

Rutaecarpine is a major quinazolinocarboline alkaloid isolated from Evodia rutaecarpa. It was reported to possess a wide spectrum of pharmacological activities, such as vasodilation, antithrombosis, and anti-inflammation. In the present study, adverse effects of rutaecarpine on immune functions were determined in female BALB/c mice. Rutaecarpine had no effects on hepatotoxicity parameters in mice, as measured by serum activities of aminotransferases. Meanwhile, rutaecarpine significantly decreased the number of antibody-forming cells and caused weight decrease in spleen in a dose-dependent manner, when mice were administered with rutaecarpine at 10mg/kg, 20mg/kg, 40 mg/kg or 80 cmg/kg once intravenously. In addition, rutaecarpine administered mice exhibited reduced splenic cellularity, decreased numbers of total T cells, CD4(+) cells, CD8(+) cells, and B cells in spleen. IL-2, interferon-gamma and IL-10 mRNA expressions were suppressed significantly by rutaecarpine treatment. The number of CD4(+)IL-2(+) cells was reduced significantly following administration of mice with rutaecarpine. Furthermore, rutaecarpine caused the cell cycle arrest in G(0)+G(1) phase in a dose-dependent manner. Rutaecarpine caused significant inductions of hepatic cytochrome P450 (CYP) 1A, 2B, and 2E1 activities dose-dependently. In the splenic lymphocyte proliferation assay, rutaecarpine inhibited proliferation by LPS and Con A ex vivo, whereas it had no effects on in vitro proliferation. These results suggested that a single bolus intravenous injection of rutaecarpine from 20mg/kg might cause immunosuppressive effects, and that rutaecarpine-induced immunosuppression might be mediated, at least in part, through the inhibition of cytokine production and cell cycle arrest in G(0)+G(1) phase, and caused possibly by mechanisms associated with metabolic activation.