玳玳果黄酮降脂滴丸的肠吸收动力学特性

目的：考察玳玳果黄酮降脂滴丸的肠吸收特性并探讨其作用机制。方法：采用大鼠外翻肠囊模型,建立同时测定大鼠肠吸收液中玳玳果黄酮降脂滴丸特征药效成分新橙皮苷和柚皮苷含量的液-质联用（UPLC-MS）分析法,比较玳玳果黄酮降脂滴丸与原料玳玳果黄酮降脂提取物在大鼠不同肠段的累积吸收量,分析不同质量浓度大鼠空肠段的吸收过程,探讨其肠吸收部位及机制。结果：玳玳果黄酮降脂滴丸特征药效成分新橙皮苷和柚皮苷90 min时的累积吸收量由多到少依次是空肠、十二指肠、回肠和结肠;玳玳果黄酮降脂滴丸在高、中、低剂量下空肠的累积吸收量随时间的增加而增加,相关系数r>0.95,且随着药液质量浓度的增加新橙皮苷和柚皮苷的Ka均增加,符合一级药动学过程;在等剂量给药下,玳玳果黄酮降脂滴丸中新橙皮苷和柚皮苷的累积吸收量约是玳玳果黄酮降脂提取物的1.6倍。结论：玳玳果黄酮降脂滴丸在全肠道均有吸收,小肠的上中段为最佳吸收部位,新橙皮苷和柚皮苷吸收可能为被动吸收循环入体,制成的玳玳果黄酮降脂滴丸可显著改善提取物在肠道的吸收,提高其口服生物利用度。     

玳玳果黄酮降脂提取物体内效应组分排泄动力学研究

目的 建立同时测定大鼠尿液及粪便中提取物效应组分新橙皮苷和柚皮苷含量方法,研究大鼠口服玳玳果黄酮降脂提取物后的尿药排泄动力学及排泄特征。方法 采用UPLC-MS/MS建立效应组分新橙皮苷及柚皮苷在大鼠尿液及粪便的定量分析方法,计算口服玳玳果黄酮降脂提取物后不同时间点新橙皮苷及柚皮苷在尿液及粪便中的排泄率,并以亏量法计算尿液中的消除半衰期及消除速率常数,评价大鼠口服玳玳果黄酮降脂提取物后尿药排泄动力学及排泄特征。结果 所建立的UPLC-MS/MS定量分析方法专属性良好、标准曲线及线性范围良好,方法准确度与精密度、定量下限均符合有关规定,该方法能够满足大鼠尿液及粪便中效应组分的定量检测需要;口服72 h后,大鼠尿液中新橙皮苷、柚皮苷的平均累计排泄率分别为（1.76±0.76）‰和（1.39±0.57）‰;大鼠粪便中新橙皮苷、柚皮苷的平均累计排泄率分别为（52.45±6.30）%和（51.57±4.80）%;口服后效应组分新橙皮苷及柚皮苷在尿液及粪便中的排泄量分别在24~36 h和4~8 h达到峰值;72 h后仍有药物经尿液排泄,给药后24 h粪便累计排泄率便达坪值;亏量法计算得新橙皮苷消除速率常数为（0.080±0.021）·h^-1,消除半衰期为（9.41±3.22）h,柚皮苷消除速率常数为（0.077±0.017）·h^-1,消除半衰期为（9.51±2.97）h,新橙皮苷及柚皮苷的动力学参数间差异无统计学意义。结论 口服给药后效应组分原形即通过粪便较快地排出体外,粪便排泄是效应组分排出体外的主要途径,新橙皮苷及柚皮苷经尿液排泄的特征无明显差异。     

玳玳果黄酮提取物关键效应组分辨识及调控脂质代谢作用机制研究

目的 探究玳玳果黄酮提取物及其敲出目标组分对脂肪变性细胞调控脂质代谢作用及机制，辨识关键效应组分。方法 采用油酸诱导建立HepG2脂肪变性细胞模型，非诺贝特为阳性对照，油红O染色法及甘油三酯(triglycerides，TG)和总胆固醇(total cholesterol，TC)为评价指标，考察玳玳果黄酮提取物、柚皮苷(目标组分)、新橙皮苷(目标组分)、其他相关组分(黄酮提取物敲除柚皮苷、新橙皮苷)、黄酮提取物敲除其他相关组分(柚皮苷-新橙皮苷6︰ 7)对脂肪变性模型细胞脂质集聚影响。基于AMPK/SREBP-1c及PPARα信号通路，采用qRT-PCR法、Western blotting检测大鼠肝脏相关基因、蛋白的表达。结果 预防性给药后，与模型组相比，玳玳果黄酮提取物、柚皮苷、新橙皮苷、提取物敲除其他相关组分的细胞脂质积聚、TG及TC含量均有明显的改善(P<0.01或P<0.05)；玳玳果黄酮提取物及柚皮苷、新橙皮苷可使促腺苷酸激活蛋白激酶(ATP-activated protein kinase，AMPK)的mRNA及磷酸化蛋白表达水平明显升高(P<0.01或P<0.05)；显著抑制并明显下调固醇调节元件结合蛋白(sterol regulatory element binding protein，SREBP-1c)、脂肪酸合成酶(fatty acidsynthase，FAS)、乙酰辅酶羧化酶(coenzyme carboxylase，ACC)的mRNA和蛋白表达水平(P<0.01或P<0.05)；可促进过氧化物酶增殖物激活受体(peroxidase proliferator-activated receptor ，PPARα)、肉毒碱棕榈酰基转移酶(carnitinepalmitoyltransferase-1，CPT-l)的mRNA及蛋白表达水平明显上升(P<0.01或P<0.05)；其他相关组分无明显差异。结论 玳玳果黄酮提取物及其关键效应组分柚皮苷、新橙皮苷可有效改善脂质积聚；其作用机制为激活AMPK促其蛋白磷酸化表达，从而抑制脂质合成，同时促进脂肪酸的氧化分解，由此减少脂肪沉积，发挥调控脂质代谢的作用。     

玳玳黄酮提取物的质量标准研究

目的：建立玳玳黄酮提取物的质量标准。方法：采用薄层色谱法鉴别玳玳黄酮提取物中的柚皮苷和新橙皮苷;采用紫外分光光度法测定玳玳黄酮提取物中总黄酮含量;采用高效液相色谱法同时检测玳玳黄酮提取物中柚皮苷和新橙皮苷的含量。结果：薄层色谱斑点清晰,易于识别,专属性强;紫外分光光度法新橙皮苷在4．44～26．64μg·mL-1范围内呈良好线性关系,平均加样回收率为99．92％（RSD=1．65％）;HPLC法柚皮苷在1.988—13．916μg·mL-1范围内呈良好线性关系,平均加样回收率为99．73％（RSD=1．56％）,新橙皮苷在1．992～13．944μg·mL-1范围内呈良好线性关系,平均加样回收率为99．58％（RsD=1．89％）。结论：所建方法操作简便,结果准确,可有效控制玳玳黄酮提取物的质量。     

玳玳黄酮滴丸质量分析

目的 建立玳玳黄酮滴丸质量评价方法。方法 采用薄层色谱法鉴别玳玳黄酮滴丸,并对玳玳黄酮滴丸进行重金属及砷盐的检查;建立高效液相色谱法同时测定玳玳黄酮滴丸主要药效成分柚皮苷和新橙皮苷含量的方法。结果 薄层色谱主要成分斑点清晰可见,阴性对照无干扰;玳玳黄酮滴丸重金属及砷盐的限量均小于百万分之十;新橙皮苷在4.088--16.352μg.mL 1内呈良好线性关系,柚皮苷在4.084--16.336μg.mL 1内呈良好线性关系,3批玳玳黄酮滴丸新橙皮苷平均含量分别为75.32,74.82,73.76 mg·g-1,柚皮苷平均含量分别为63.05,63.21,62.07mg·g-1。结论 所建立的鉴别、检查、含量测定方法稳定简便,可有效控制玳玳黄酮滴丸质量。     

玳玳黄酮自微乳化微丸的体外溶出性

目的:研究玳玳黄酮自微乳化微丸中柚皮苷和新橙皮苷的体外溶出度。方法:运用HPLC法测定玳玳黄酮自微乳化微丸在纯化水中柚皮苷和新橙皮苷的溶出度,分别用零级动力学方程、一级动力学方程和Higuchi方程对溶出数据进行拟合,并采用相似因子(f2)分析溶出曲线。结果:玳玳黄酮自微乳化微丸溶出过程以一级动力学方程拟合最佳。柚皮苷和新橙皮苷溶出曲线的f2为92.48。结论:玳玳黄酮自微乳化微丸中柚皮苷和新橙皮苷在体外释放动力学符合一级动力学方程,能够从自微乳化微丸中同歩释放。     

小檗碱与枳实提取物配伍在大鼠体内的药动学研究

目的 建立UPLC-MS-MS生物样品分析方法,同时测定大鼠血浆中小檗碱、柚皮苷、橙皮苷与新橙皮苷,测定SD大鼠ig小檗碱和枳实提取物后血药浓度及相关药动学参数的变化。方法 以UPLC Acquity BEH C18 （50 mm×2.1 mm,1.7μm）为色谱柱,含0.05%甲酸2 mmol/L甲酸铵水（水相A）-含0.05%甲酸乙腈（有机相B）为流动相梯度洗脱;SD大鼠分为ig小檗碱组、枳实提取物组、小檗碱和枳实提取物配伍组。结果 UPLC-MS/MS可快速分析血浆样品中小檗碱、柚皮苷、橙皮苷与新橙皮苷,方法学验证符合生物样品分析方法要求;SD大鼠ig小檗碱与枳实提取物配伍后,小檗碱血药浓度较单给小檗碱组明显提高,生物利用度明显提高;同时大鼠血浆中可检测到柚皮苷与新橙皮苷。结论 SD大鼠给药小檗碱后,枳实黄酮的生物利用度明显提高,提示小檗碱与枳实黄酮有明显的药物相互作用。     

玳玳果黄酮滴丸的溶出度及速释机制

目的:评价玳玳果黄酮滴丸的溶出度并初步探讨其速释机制。方法:在多种pH值溶出介质中采用多条溶出曲线评价玳玳果黄酮滴丸的体外溶出度,以柚皮苷为指标成分,运用HPLC法考察玳玳果黄酮滴丸在不同时间的累积溶出百分数,并通过差示扫描热分析研究滴丸的速释机制。结果:4种释放介质中柚皮苷在2.12～53.00mg.L-1范围内线性关系良好,玳玳果黄酮滴丸中柚皮苷在20 min时累积溶出度可达85%。差示扫描量热分析结果显示,玳玳果黄酮在PEG4000-PEG6000中形成了某种形式的固体分散体。结论:玳玳果黄酮滴丸具有良好的溶出度,玳玳果黄酮有效部位在滴丸内形成了良好的固体分散体,使药物在基质中的分散度增加而达到速释。     

玳玳黄酮自微乳化软胶囊制备过程药效成分转移率考察

目的 针对玳玳总黄酮有效部位多组分特性,基于HPLC特征图谱研究玳玳黄酮自微乳化软胶囊制备过程中的药效成分转移率。方法 采用HPLC特征图谱法,色谱柱为Lichrocart C₁₈(250 mm×4.0 mm,5 μm);流动相为甲醇-0.1%的磷酸水溶液(梯度洗脱);流速1.0 mL·min^-1;检测波长284 nm;柱温30 ℃。结果 玳玳黄酮自微乳化软胶囊制备工艺过程中8个共有峰的转移率为80.55%~95.93%,其中主要特征成分柚皮苷和新橙皮苷迁移率均>95%,各药效成分群整体迁移相似度>0.999。结论　玳玳黄酮自微乳化软胶囊制备工艺能够比较完整的保留玳玳总黄酮提取物的整体药效部位成分群。     

玳玳黄酮自微乳化微丸含量测定及HPLC特征图谱分析

目的:建立玳玳黄酮自微乳化微丸中柚皮苷、新橙皮苷HPLC含量测定方法和HPLC特征图谱,为玳玳黄酮自微乳化微丸的质量控制提供有效的方法。方法:含量测定采用Agilent-C18色谱柱(250 mm×4.6 mm,5μm),流动相为乙腈-0.1%磷酸水溶液(22∶78),流速为1.0 mL.min-1,柱温为室温。HPLC特征图谱测定采用250-4 Lichrocart C18色谱柱(250 mm×4.0mm,5μm),流动相为甲醇-乙腈-0.1%的磷酸水溶液,梯度洗脱,流速1.0 mL.min-1,柱温30℃。检测波长均为283 nm。结果:柚皮苷浓度在4.48～17.92μg.mL-1范围内线性关系良好(r=0.9998),平均加样回收率为97.5%(RSD=1.0%);新橙皮苷浓度在5.92～23.68μg.mL-1范围内线性关系良好(r=0.9997),平均加样回收率为98.8%(RSD=0.82%)。在此条件下所建立的特征图谱中各成分得到有效分离,各批玳玳黄酮自微乳化微丸的相似度在0.900～1.000间。结论:HPLC法同时测定玳玳黄酮自微乳化微丸中柚皮苷和新橙皮苷含量的方法简便、准确,建立的HPLC特征图谱可用于评价微丸的物质组成及有效物质群的质量变化,可作为玳玳黄酮自微乳化微丸质量控制的依据。     

Pharmacokinetics and tissue distribution of gentiopicroside following oral and intravenous administration in mice.

The pharmacokinetics and tissue distribution of Gentiopicroside (GPS), one of the major active components of the Gentiana species of medicinal plants, was studied following oral and intravenous administration in mice. The distribution of GPS in mice after oral and intravenous doses could be fitted to a two-compartments open model. The serum half-life of GPS was 6.1 h and 2.8 h for intravenous and oral administration, respectively. The Tmax of GPS after oral administration was 0.50 h, and the bioavailability was 39.6%. The AUC gradient in individual tissues following intravenous administration was kidney >serum >liver >spleen >lung >thymus >fat >heart >muscle >stomach >intestinal >brain. The MRT gradient was muscle >serum >lung >spleen >lung >intestinal>heart >stomach >brain >liver >thymus >kidney >fat. Overall the data show that GPS could be absorbed rapidly in mice, but with a low bioavailability, and could distribute to tissues extensively, but was generally cleared quickly with short MRTs. The study demonstrates the need for repeated dosage, or better, a slow release formulation as an ideal means of administering GPS.     

Transporters (OATs and OATPs) contribute to illustrate the mechanism of medicinal compatibility of ingredients with different properties in yuanhuzhitong prescription

Various medicinal ingredients with different tastes are combined according to the theory of compatibility in Chinese materia medica to achieve a better efficacy, while the mechanism was not very clear. Here, the authors studied the interaction between ingredients and human transporters such as the kidney transporters OAT1 and OAT3, the liver transporters OATP1B1 and OATP1B3, and the intestine transporter OATP2B1 to discern the compatibility mechanism of ingredients with different tastes in the Yuanhuzhitong preparation (YHP) comprising Corydalis yanhusuo (CYH) and Angelica dahurica (AD), which could relieve pain by restraining the central system. The results show that tetrahydropalmatine (TDE), the major component of CYH, could be transported by OAT3 into kidney, OATP1B1 and OATP1B3 into liver, while imperatorin (IPT) and isoimperatorin (ISP), the two key components of AD, and AD extract showed strong inhibition to OAT1 and OAT3. What's more, AD extract also exerted strongly inhibition to human transporters OATP1B1 and OATP1B3. It was also detected that IPT, ISP, and AD extract significantly downregulated the expression of Oatp1a1, Oatp1a4, and Oatp1b2 of liver in mice. The in vivo results show that the concentration of TDE in liver and kidney significantly decreased, while the TDE concentration in blood and brain were both significantly enhanced in the presence of IPT, ISP, and AD extract. These results suggest that the ingredients in AD with pungent taste could enhance the exposure of TDE in blood and brain by inhibiting the uptake of TDE in liver and kidney. That is to say, TDE with bitter taste could “flood up” into the central nervous system to play its therapeutic effect by the cut-off of that into liver and kidney in the presence of ingredients within AD. This paper not only proves the meridian distribution of CYH in liver and kidney with the role of OAT3, OATP1B1, and OATP1B3, but also illustrates how to improve the efficacy of CYH by reasonable compatibility with AD. This study may offer a valuable clue to illustrate the mechanism of compatibility theory.     

Formulating Paclitaxel in Nanoparticles Alters Its Disposition

Purpose Paclitaxel is active and widely used to treat multiple types of solid tumors. The commercially available paclitaxel formulation uses Cremophor/ethanol (C/E) as the solubilizers. Other formulations including nanoparticles have been introduced. This study evaluated the effects of nanoparticle formulation of paclitaxel on its tissue distribution.Methods We compared the plasma and tissue pharmacokinetics of paclitaxel-loaded gelatin nanoparticles and the C/E formulation. Mice were given paclitaxel-equivalent doses of 10 mg/kg by intravenous injection.Results The nanoparticle and C/E formulations showed significant differences in paclitaxel disposition; the nanoparticles yielded 40% smaller area under the blood concentration-time curve and faster blood clearance of total paclitaxel concentrations (sum of free, protein-bound, and nanoparticle-entrapped drug). The two formulations also showed different tissue specificity. The rank order of tissue-to-blood concentration ratios was liver > small intestine > kidney >> large intestine > spleen = stomach > lung > heart for the nanoparticles, and liver > small intestine > large intestine > stomach > lung kidney > spleen > heart for the C/E formulation. The nanoparticles also showed longer retention and higher accumulation in organs and tissues (average of 3.2 ± 2.3-fold), especially in the liver, small intestine, and kidney. The most striking difference was an 8-fold greater drug accumulation and sustained retention in the kidney.Conclusions These data indicate that formulation of paclitaxel affects its clearance and distribution into tissues, with preferential accumulation of nanoparticles in the liver, spleen, small intestine, and kidney.     

香加皮不同组分大鼠长期毒性研究

目的 观察连续给予香加皮不同组分导致大鼠慢性毒性的损伤表现、程度及可逆性.方法 分别给140只大鼠灌胃高、中、低剂量的香加皮水提组分和醇提组分样品,除观察一般状况外,检测血常规、血生化指标,剖杀大鼠,精密称取心、肝、脾、肺、肾脏等重要脏器,计算脏、体比值,进行常规病理检查.停药后,进行恢复期观察.结果 连续20天给予不同剂量的香加皮水提组分和连续9天给予不同剂量的醇提组分样品均可导致大鼠体重下降,饮食、饮水不佳,血ALT、AST、AKP、TPC增高,ALB、CR降低、A/G比值降低,肝脏重量和肝体比值、肾脏重量和肾体比值增大,病理检查可见不同程度的肝脏、肾脏病理组织损伤;对血常规影响不明显;肝、肾毒性损伤程度与给药剂量呈现一定的剂量依赖相关性;经过恢复期观察,上述部分病变不可逆.结论 香加皮水提组分给药20天、醇提组分给药9天造成的慢性毒性损伤部位以肝、肾损伤为主,且部分病变为不可逆性损伤.     

UPLC-MS/MS法测定氘代氯化琥珀胆碱在大鼠体内分布

目的:建立超高效液相色谱串联质谱(UPLC-MS/MS)法测定氘代氯化琥珀胆碱(2H-Suc)在大鼠主要脏器分布。方法:大鼠皮下注射2H-Suc 4 h后,用UPLC-MS/MS法测定血清和相关脏器中2H-Suc含量;色谱柱:Luna NH2色谱柱(2 mm×100 mm,3μm),流动相为0.1%甲酸水溶液-乙腈等度洗脱,流速为0.2 mL/min;正离子扫描模式,多反应监测模式下,电喷雾离子化源进行检测;考察该方法的线性关系与检测限、准确度和精密度、回收率、基质效应和稳定性。结果:血清和肾组织中2H-Suc的检测限分别是0.04 ng/mL和0.05 ng/mL,线性范围分别是0.3~100 ng/mL和0.5~100 ng/mL,日内、日间精密度和准确度均小于15%,回收率在85.30%~96.76%之间,经过处理后的样品不存在明显的基质效应。大鼠皮下注射2H-Suc,在血清及心、肝、肾等主要脏器中均检测到2H-Suc,其中肾脏中含量最高。结论:UPLC-MS/MS法是测定大鼠体内2H-Suc含量的一个有效方法,2H-Suc在大鼠血清及心、肝、肾等中均有分布,肾脏中含量最高。     

Pharmacokinetics, tissue distribution, metabolism, and excretion of depside salts from Salvia miltiorrhiza in rats.

Salviae miltiorrhiza, a traditional Chinese medical herb known as "Danshen," has been widely used in clinics to improve blood circulation, relieve blood stasis, and treat coronary heart disease. Depside salts from S. miltiorrhiza are a novel drug in which magnesium lithospermate B and its analogs are the active components. The pharmacokinetics, tissue distribution, metabolism, and excretion of three of the major components, lithospermic acid B, rosmarinic acid (RA), and lithospermic acid (LA), were studied by liquid chromatography-tandem mass spectrometry following intravenous administration in Sprague-Dawley rats. The elimination half-lives for LSB, RA, and LA were 1.04, 0.75, and 2.0 h, respectively, when 60 mg/kg S. miltiorrhiza depside salts were administrated. The areas under the curve for LSB, RA, and LA were 51.6, 6.6, and 25.2 mg . h/l, respectively, and the values decreased in the individual tissues in the following order: kidney > lung > liver > heart > spleen > brain for LSB; kidney > lung > heart > liver > spleen > brain for RA; and heart > lung > kidney > liver > spleen > brain for LA. After intravenous administration of 60 mg/kg S. miltiorrhiza depside salts, 86% of the LSB was excreted in the bile within 6 h. The main metabolites M1 and M2 were found in the serum. Overall, the results show that depside salts from S. miltiorrhiza are rapidly and widely distributed to tissues after intravenous administration in rats but that they are also rapidly cleared and excreted.