草乌豆腐法炮制工艺研究

目的 探索最佳的豆腐法炮制草乌的工艺参数。方法 采用L₁₆(4₃)正交试验方法,以浸漂时间,水煮时间和豆腐用量三因素,各设置四水平进行16次试验,每次实验结果均以单酯型和双酯型生物碱总量测定数据为评判指标。结果 浸漂时间四水平四次试验结果的单酯型生物碱含量分别为0.11%、0.07%、0.062%和0.048%;水煮时间四次结果的含量分别为0.081%、0.066%、0.074%和0.067%;豆腐用量四次结果的含量分别为0.070%、0.072%、0.080%和0.073%;双酯型生物碱16次测定结果均小于0.034%。结论 三因素中对单酯型生物碱含量影响大小的次序为浸漂时间>水煮时间>豆腐用量。在上述任何条件下,毒性成分双酯型生物碱均小于中国药典的限度要求,均符合规定,而毒性成分双酯型生物碱总量随豆腐用量的增加而明显减少。豆腐法炮制草乌的最佳工艺条件为A3B3C2,即浸漂6天,水煮5小时,加豆腐25%。     

藏药榜嘎中的双酯型和内酯型生物碱分析方法研究

目的建立藏药榜嘎中双酯型和内酯型生物碱的测定方法,为榜嘎的质量标准提高提供参考。方法采用高效液相色谱法。色谱柱为:Phenomenes luna C18(250mm×4.6mm,5μm);流速:1.0mL·min-1。双酯型生物碱的流动相为:A为乙腈-四氢呋喃(25∶15),B为0.1mol·L-1醋酸铵溶液(每1 000mL加冰乙酸0.5mL),梯度洗脱;检测波长为235nm;柱温为室温。内酯型生物碱的流动相为甲醇-水-氯仿-三乙胺(70∶30∶2∶0.1);检测波长230nm;柱温30℃。结果榜嘎中均不含新乌头碱、乌头碱、次乌头碱等双酯型生物碱;内酯型生物碱:阿替新在2.024²0.24μg范围内线性关系良好,线性方程:Y=1.918×104 X-678(r=0.999 9),平均回收率97.3%,RSD=0.9%;异叶乌头碱在2.00820.24μg范围内线性关系良好,线性方程:Y=1.918×104 X-678(r=0.999 9),平均回收率97.3%,RSD=0.9%;异叶乌头碱在2.008²0.08μg范围内线性关系良好,线性方程:Y=1.602×104 X-215(r=0.999 9),平均回收率98.6%,RSD=1.4%。结论该方法准确,可靠,专属性强,可用于榜嘎的质量控制。     

Involvement of an Orphan Transporter,SLC22A18, in Cell Growth and Drug Resistance of Human Breast Cancer MCF7 Cells

The SLC22A18 gene, which encodes an orphan transporter, is located at the 11p15.5 imprinted region, an important tumor suppressor gene region. However, the role of SLC22A18 in tumor suppression remains unclear. Here, we investigated the involvement of SLC22A18 in cell growth, invasion, and drug resistance of MCF7 human breast cancer cell line. Western blot analysis indicated that SLC22A18 is predominantly expressed at intracellular organelle membranes. Quantitative proteomics showed that knockdown of SLC22A18 significantly altered the expression of 578 (31.0%) of 1867 proteins identified, including proteins related to malignancy and poor prognosis of breast cancer. SLC22A18 knockdown (1) increased MCF7 cell growth concomitantly with a >7-fold increase of annexin A8 (involved in cell growth and migration; a predictor of poor prognosis), (2) induced spherical morphology of MCF7 cells concomitantly with a nearly 3-fold increase of CD44 (involved in regulation of malignant phenotypes), and (3) increased chemosensitivity to vinca alkaloids concomitantly with a >80% reduction of doublecortin-like kinase 1 (involved in regulation of microtubule polymerization). Our results suggest that SLC22A18 may act as a tumor suppressor by regulating the expression levels of cell growth–related proteins, and vinca alkaloids might show therapeutic efficacy against low-SLC22A18–expressing breast cancer.     

星点设计-响应面法优化川芎生物碱提取工艺

目的优选川芎总生物碱的提取工艺。方法对川芎采用乙醇回流提取的方法,进行星点设计,以乙醇浓度、醇倍数、提取时间为自变量,以川芎中总生物碱含量为因变量,通过对自变量各水平的多元线性回归及二项式拟合,用响应面法优选出最佳提取工艺,同时进行预测分析。结果确定了最佳提取工艺为用8倍90%的乙醇提取2次,每次2 h,提取的川芎总生物碱含量为0.693 mg.g^-1,二项式拟合复相关系数R^2=0.957 3。结论星点设计-响应面法优选川芎中总生物碱的提取工艺,方法简单,精密度高,可预测性好。     

异紫堇二酮体内外抗肿瘤活性研究

目的异紫堇二酮的体内外抗肿瘤活性及其作用机制的初步研究。方法通过异紫堇二酮对体外培养肿瘤细胞的生长抑制,体内荷S180瘤和荷H22瘤昆明种小鼠的肿瘤生长抑制、体重以及对小鼠免疫器官的影响(脾脏指数、胸腺指数),对其抗肿瘤活性进行综合评价,并通过流式细胞术检测异紫堇二酮作用下肺腺癌细胞(A549)的细胞凋亡及周期分布情况,初步探讨该化合物的抗肿瘤作用机制。结果异紫堇二酮对9种所选肿瘤细胞具有一定的生长抑制作用,IC50值在1.452×104～2.460×104 mol.L-1之间;流式细胞术检测结果显示异紫堇二酮可诱导A549细胞凋亡,使A549细胞阻滞于G0/G1期并随作用时间延长出现明显的凋亡峰;剂量为100和200 mg.kg-1.d-1时,异紫堇二酮对荷S180瘤和荷H22瘤小鼠的抑瘤率均大于40%,但剂量在200 mg.kg-1.d-1时,荷瘤小鼠的胸腺指数与脾脏指数相比较生理盐水组均有明显降低。结论异紫堇二酮具有一定的抗肿瘤活性,其主要是阻断肿瘤细胞由G0/G1期向S期转变来诱导肿瘤细胞死亡。     

大环双内酯雷公藤生物碱高分辨质谱数据库的建立与应用

目的:建立大环双内酯雷公藤生物碱高分辨质谱数据库(HR-MS-database),并应用于不同种属雷公藤生物碱的快速鉴定分析.方法:采用超高效液相色谱-四极杆-飞行时间质谱联用(UPLC/Q-TOF-MS)技术手段,借助SCIEX公司LibraryView数据库平台,建立大环双内酯雷公藤生物碱高分辨质谱-数据库(HR-...     

HPLC同时测定苦黄注射液中生物碱和大黄蒽醌类成分的含量

目的 建立同时测定苦黄注射液中生物碱和大黄蒽醌类成分的定量分析方法。方法 采用Venusil MP C₁₈(2)(4.6 mm×250 mm,5 μm)色谱柱为分析柱,甲醇(A)-0.4%二乙胺(B),梯度洗脱(A为5%→5%→40%→60%→80%→5%,相应时间周期为0→5→20→30→55→60 min),流速为1.0 mL·min^-1,紫外检测波长为254 nm,柱温位30 ℃。结果 在选定色谱条件下,芦荟大黄素、大黄酸、苦参碱、大黄素、大黄酚、氧化苦参碱、大黄素甲醚、槐定碱、槐果碱分别在0.012 3~0.369?μg、0.023~0.69 μg、0.138~4.14 μg、0.056~1.68 μg、0.065~1.95 μg、0.145~4.35 μg、0.012?5~0.375 μg、0.146~4.38 μg和0.101~ 3.03 μg内线性关系良好(r=0.999 3,0.999 6,0.999 6,0.999 6,0.999 4,0.999 3,0.999 5,0.999 4,0.999 4),加样回收率分别为98.38%,100.79%,98.03%,99.81%,98.62%,99.28%,98.90%,101.75%,100.75%,RSD分别为1.60%,2.65%,2.12%,2.78%,2.08%,2.45%,2.08%,2.35%,1.72%。结论 该方法操作简便易行,重复性好,结果准确可靠,可为苦黄注射液的质量控制提供定量评价方法。     

附子心脏毒性及配伍减毒增效机制研究进展

附子来源于毛茛科植物乌头Aconitum carmichaeli Debx.,最主要的毒性反应表现为心脏毒性和神经毒性,其中心脏是主要的靶器官之一。目前普遍认为附子所致心脏毒性是由双酯型二萜生物碱直接或间接作用于心脏的结果,其毒性机制与Ca²⁺超载、脂质过氧化反应、细胞凋亡以及迷走神经兴奋性增高有关。研究发现附子与其他传统中草药配伍能通过影响体内代谢过程、抗Ca²⁺超载、抗氧化应激、抗凋亡等机制发挥减毒增效作用。就近年来国内外对附子心脏毒性及配伍减毒增效相关机制的研究进展进行综述,以期为附子的深入研究和临床安全用药提供参考。     

Study on the pharmacokinetics,tissue distribution and excretion of laurolitsine from Litsea glutinosa in Sprague-Dawley rats

ContextLaurolitsine is an aporphine alkaloid and exhibits potent antihyperglycemic and antihyperlipidemic effects in ob/ob mice.ObjectiveTo investigate the pharmacokinetics, tissue distribution and excretion of laurolitsine.Materials and methodsA LC-MS/MS method was established and validated to determine laurolitsine concentrations in the biological matrix of rats (plasma, tissue homogenate, urine and faeces). 10 Sprague-Dawley (SD) rats were used for plasma exposure study: 5 rats were injected with 2.0 mg/kg of laurolitsine via the tail vein, and the other 5 rats were administered laurolitsine (10.0 mg/kg) by gavage. 25 SD rats used for tissue distribution study and 5 SD rats for urine and faeces excretion study: rats administered laurolitsine (10.0 mg/kg) by gavage. After administered, serial blood, tissue, urine and faeces were collected. Analytical quantification was performed by a previous LC-MS/MS method. The pharmacokinetics, bioavailability, tissue distribution and excretion of laurolitsine were described.ResultsThe pharmacokinetic parameters of oral and intravenous administration with T_max were 0.47 and 0.083 h, t_1/2 were 3.73 and 1.67 h, respectively. Oral bioavailability was as low as 18.17%. Laurolitsine was found at a high concentration in the gastrointestinal tract, liver, lungs and kidneys (26 015.33, 905.12, 442.32 and 214.99 ng/g at 0.5 h, respectively) and low excretion to parent laurolitsine in urine and faeces (0.03 and 1.20% in 36 h, respectively).ConclusionsThis study established a simple, rapid and accurate LC-MS/MS method to determine laurolitsine in different rat samples and successful application in a pharmacokinetic study.     

十二种莨菪碱类衍生物药理作用的比较

本研究用放射配基结合法测定了十二种莨菪碱类衍生物对豚鼠回肠纵长肌M胆碱受体的亲和力,并测定了其油水分布系数和对动物记忆获得的影响。结果表明,大部分衍生物季铵盐化后可降低与受体的亲和力和油水分布系数,但是托品酸α位碳原子上带有一个羟基的季铵盐如8610等,其与受体的亲和力分别比东莨菪碱和正丁基东莨菪碱强10倍和100倍,而油水分布系数却比东莨菪碱和阿托品小。十二种莨菪碱类衍生物阻抑小鼠记忆获得所需剂量比东莨菪碱大2.5到5倍不等。