RP-HPLC同时测定复方三七制剂中的人参皂苷Rg1、Rb1

目的 建立复方三七制剂中人参皂苷Rg1、Rb1含量测定的方法.方法 采用RP-HPLC法,色谱柱为Phenomenex C18(250 mm x4.6 mm,5 μm);流速1.0 ml·min-1;检测波长203 nm;柱温30℃;流动相为乙腈-水(梯度洗脱).结果 人参皂苷Rg1的线性范围为0.2124～2.1240 μg(r=0.9999),平均回收率为100.24%,RSD=1.14%(n=9);人参皂苷Rb1的线性范围为0.2108～2.1080μg(r=0.9999),平均回收率为99.71%,RSD=0.79%(n=9).结论 所建方法简便、准确,重复性好,可同时测定复方三七制剂中人参皂苷Rg1和人参皂苷Rb1的含量.     

RP-HPLC梯度洗脱法测定三七胶囊中三七皂苷R1和人参皂苷Rb1及人参皂苷Rg1的含量

目的:研究三七胶囊中三七皂苷R1、人参皂苷Rb1及人参皂苷Rg1的含量测定方法,为制定质量标准中含量测定方法及含量限度提供依据。方法:用岛津Shim-packC18分析柱,乙腈-水梯度洗脱,0～6 min(24:76),6～15 min(27:73→40:60),流速:1.0 ml.min-1,检测波长:203 nm,柱温:35℃,进样量:20μl。结果:三七皂苷R1在0.38～3.80μg范围内呈良好的线性关系(r=0.9997),平均回收率为100.3%,RSD为1.08%;人参皂苷Rb1在1.50～15.00μg范围内呈良好的线性关系(r=0.9993),平均回收率为99.9%,RSD为1.03%;人参皂苷Rg1在1.46～14.60μg范围内呈良好的线性关系(r=0.9992),平均回收率为99.8%,RSD为1.03%。结论:该方法能将多种皂苷很好的分离检测,而且方法简便、快速、重现性好、准确可靠,可用于三七胶囊的质量控制。     

高效液相色谱梯度洗脱法同时测定三七总皂苷中人参皂苷Rb1、人参皂苷Rg1和三七皂苷R1含量

目的 采用高效液相色谱梯度洗脱法测定三七总皂苷中人参皂苷Rb1、人参皂苷Rg1和三七皂苷R1的含量。方法 色谱柱为C18柱（250 mm×4.6 mm,3.5μm）,流动相为乙腈-水、梯度洗脱,检测波长为203nm,流速1.0 mL/min,柱温为室温。结果 人参皂苷Rg1进样量线性范围是6.0~18μg（r=0.996 7）,平均回收率为98.79%,RSD=0.33%（n=6）;人参皂苷Rb1进样量线性范围是6.0~18μg（r=0.996 4）,平均回收率为98.64%,RSD=0.46%（n=6）;三七皂苷R1进样量线性范围是1.6~4.8μg（r=0.997 1）,平均回收率为98.84%,RSD=0.56%（n=6）。结论 该方法准确、灵敏度高,可用于三七总皂苷的质量控制。     

高效液相色谱法测定三七伤药片中3组分的含量

目的建立以高效液相色谱法测定三七伤药片中三七皂苷R1和人参皂苷Rg1、Rb1含量的方法。方法色谱柱为VP-ODS(150mm×4.6mm,5μm),使用梯度洗脱系统,流动相为乙腈-水(0~20min(20∶80),20~26min(40∶60),26~35min(20∶80)),流速为1.0mL.min-1,柱温为室温,检测波长为203nm。结果三七皂苷R1的线性范围为0.261~7.83μg(r=0.9999),平均回收率为(101.12±2.03)%(RSD=2.1%);人参皂苷Rg1的线性范围为0.6025~18.075μg(r=0.9997),平均回收率为(97.22±3.33)%(RSD=1.9%);人参皂苷Rb1的线性范围为0.5375~16.125μg(r=0.9996),平均回收率为(100.71±2.4)%(RSD=1.6%)。结论该法可用于测定三七伤药片中三七皂苷R1、人参皂苷Rg1、Rb1的含量。     

高效液相色谱法测定复方三七片中人参皂苷Rg1含量

目的:用高效液相色谱法测定复方三七片中人参皂苷Rg1含量.方法:以DOS C18柱为固定相,以乙腈-0.05%磷酸溶液(20:80)为流动相,检测波长为223 nm,流速为1.4 mL/min,柱温为25℃.结果:人参皂苷Rg1的浓度线性范围是1.02～5.10μg/mL,r=0.999 7;样品平均回收率为95.67%,RSD=3.62%.结论:高效液相色谱法简便,结果准确,重现性好,可作为复方三七片中人参皂苷Rg1的含量检测方法.     

HPLC法同时测定伤科七味片中人参皂苷Rb1、人参皂苷Rg1及三七皂苷R1的含量

目的:建立伤科七味片(三七、红花等)中人参皂苷Rb1、人参皂苷Rg1及三七皂苷R1的含量测定方法。方法:采用Zobax C18色谱柱(4.6 mm×150 mm,5μm);流动相以乙腈为A,以水为B,进行梯度洗脱;检测波长:203 nm;流速:1.0 mL.min-1;柱温:30℃。结果:人参皂苷Rb1、人参皂苷Rg1、三七皂苷R1的线性范围分别为0.102 6~1.026 mg.mL-1(r=0.999 8)、0.101 4~1.014 mg.mL-1(r=0.999 9)、0.022 68~0.226 8 mg.mL-1(r=0.999 8),平均回收率分别为98.55%(RSD为0.80%)、98.63%(RSD为0.81%)、97.95%(RSD为0.97%)。结论:本方法专属、重现性好,可用于伤科七味片中人参皂苷Rb1、人参皂苷Rg1及三七皂苷R1的含量测定。     

HPLC-ELSD法同时测定复方三七漱口液中三七皂苷R_1、人参皂苷Rg_1、Rb_1的含量

目的建立同时测定复方三七漱口液中三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1的HPLCELSD法。方法采用DIKMA Diamonsil C18(2)(250 mm×4.6 mm,5μm)色谱柱,流动相为乙腈-水梯度洗脱,流速1.0 mL/min,柱温30℃,蒸发光散射检测器漂移管温度47℃,载气流速1.5 L/min。结果三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1分别在0.41¹.53、1.551.53、1.55⁵.83、1.585.83、1.58⁵.92μg呈良好线性关系;复方三七漱口液中三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1的平均回收率(n=9)分别为99.31%、100.30%、99.98%,RSD分别为1.60%、0.56%、0.97%。结论该方法简便、准确,专属性、重复性好,可用于复方三七漱口液的质量控制。     

HPLC法同时测定三七伤药胶囊中三七皂苷R_1、人参皂苷Rg_1、人参皂苷Rb_1的含量

目的:建立同时测定三七伤药胶囊中三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1含量的方法。方法:采用高效液相色谱法。色谱柱为CAPCELLPAK C18,流动相为乙腈-水(梯度洗脱),检测波长为203 nm,柱温为35℃,流速为1.0 ml/min,进样量为10μl。结果:三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1的进样量分别在0.207~4.146、0.515~10.293、0.505~10.093μg范围内与各自峰面积呈良好线性关系(r均为0.999 9);精密度试验的RSD<1.0%,稳定性、重复性试验的RSD<3%;加样回收率分别为99.40%、101.64%、101.36%,RSD分别为2.89%、2.45%、2.52%(n=6)。结论:该方法操作简便、准确性高、重复性好,可用于三七伤药胶囊中三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1的含量测定。     

HPLC法测定痛舒胶囊中三七皂苷R_1及人参皂苷Rg_1、Rb_1

目的建立液相色谱法测定痛舒胶囊中三七皂苷R1及人参皂苷Rg1、Rb1含量。方法采用WatresSymmetry RP18(4.6 mm×250 mm,5μm)色谱柱,乙腈-水为流动相,梯度洗脱,体积流量1.0 mL.min-1,检测波长203 nm,柱温30℃。结果三七皂苷R1的线性范围为0.206～3.09μg(r=0.999 4),平均加样回收率为104.2%,RSD为0.77%(n=9);人参皂苷Rg1的线性范围为0.882～13.23μg(r=0.999 5),平均加样回收率为100.7%,RSD为1.1%(n=9);人参皂苷Rb1的线性范围为0.921～13.82μg(r=0.999 2),平均加样回收率为103.1%,RSD为2.1%(n=9)。结论本方法灵敏、准确,重现性好,可用于本制剂的含量测定及质量控制。     

搜风通胶囊中三七皂苷R_1、人参皂苷Rg_1和Rb_1的含量测定

目的建立以高效液相色谱法(HPLC)测定搜风通胶囊中三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1含量的方法。方法采用色谱柱Hypersil ODS C18柱(4.6mm×250mm,5μm);流动相为乙腈(A)-水(B),进行梯度洗脱(0～12min,19%A,12～60min,19%～36%A),流速:1.0mL/min,柱温:30℃,检测波长:203nm。结果三七皂R1、人参皂苷Rg1和人参皂苷Rb1的线性范围分别为0.107～2.638μg(r=0.9998)、0.428～10.623μg(r=0.9999)和0.424～10.526μ(gr=0.9997);平均加样回收率分别为98.33%(RSD=1.16%)、98.22%(RSD=1.60%)和97.78%(RSD=0.98%)。结论本方法简便、准确、重现性好,可用于搜风通胶囊中三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1含量的测定。     

Preferential induction of CYP1A1 and CYP1B1 in CCSP-positive cells.

Both benzo[a]pyrene (BaP) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) are potent ligands of aryl hydrocarbon receptors (AhR). Although animal studies indicate that both compounds induce pathological changes in the peripheral lung, the specific cell type involved remains unclear. Clara cells, expressing Clara cell specific protein (CCSP) and abundant in cytochrome P450, are nonciliated bronchiolar epithelial cells in the peripheral lung. Here we explore the hypothesis that CCSP-positive Clara cells are highly responsive to AhR ligands and are the primary cell type involved in BaP- and TCDD-induced toxicities. The responsiveness to AhR ligands was evaluated by measuring the respective mRNA and protein levels of cytochrome P450 1A1 (CYP1A1) and 1B1 (CYP1B1) using real-time RT-PCR and immunocytochemistry assays. Two in vitro models were used: primary cultures of human small airway epithelial (SAE) cells and rat lung slice cultures. In the presence of calcium, human SAE cells differentiated into CCSP-positive cells. BaP- and TCDD-induced mRNA and protein levels of CYP1A1 and CYP1B1 levels were significantly elevated in CCSP-positive cell cultures. Similarly, AhR mRNA and protein levels were increased in CCSP-positive cell cultures, as determined by real-time RT-PCR and Western blot analysis. When rat lung slice cultures were treated with BaP or TCDD for 24 h, CYP1A1 and CYP1B1 proteins were strongly induced in Clara cells. These results indicate that, in the peripheral lung of both rats and humans, CCSP-positive cells (Clara cells) may be more sensitive to AhR ligands than other cell types.     

有机阴离子转运多肽1B1的遗传药理学进展

人体存在多种类型的药物转运体,对于药物的吸收、分布和排泄起重要作用。参与药物跨膜转运的转运体功能受影响,将可能导致诸多临床药物的疗效、毒副作用甚至药物相互作用的发生。在各种影响因素中,遗传多态性所起的作用最为重要,可导致基因表达和蛋白功能发生改变。目前,阐明转运体基因的多态性以及基因型与表型之间的相互关系已成为应用遗传信息指导临床个体化用药的必要步骤。本文就肝脏有机阴离子转运多肽1B1（OATP1B1[OATP-C],编码基因SLCO1B1）基因多态性对药代动力学和药效动力学的影响及其临床意义等方面的进展作一综述。     

Sulfation of resveratrol in human liver: Evidence of a major role for the sulfotransferases SULT1A1 and SULT1E1

Sulfation of resveratrol, a polyphenolic compound present in grapes and wine with anticancer and cardioprotective activities, was studied in human liver cytosol. In the presence of 3′-phosphoadenosine-5′-phosphosulfate, three metabolites (M1–3) whose structures were identified by mass spectrometry and NMR as trans-resveratrol-3-O-sulfate, trans-resveratrol-4′-O-sulfate, and trans-resveratrol-3-O-4′-O-disulfate, respectively. The kinetics of M1 formation in human liver cytosol exhibited an pattern of substrate inhibition with a K_i of 21.3?±?8.73?µM and a V_max/K_m of 1.63?±?0.41?µL?min^?1mg^?1 protein. Formation of M2 and M3 showed sigmoidal kinetics with about 56-fold higher V_max/K_m values for M3 than for M2 (2.23?±?0.14 and 0.04?±?0.01?µL?min^?1?mg^?1). Incubation in the presence of human recombinant sulfotransferases (SULTs) demonstrated that M1 is almost exclusively catalysed by SULT1A1 and only to a minor extent by SULT 1A2, 1A3 and 1E1, whereas M2 is selectively formed by SULT1A2. M3 is mainly catalysed by SULT1A2 and 1A3. In conclusion, the results elucidate the enzymatic pathways of resveratrol in human liver, which must be considered in humans following oral uptake of dietary resveratrol.     

抗癌药细胞色素P450 1B1抑制剂的研究进展

介绍了有关人细胞色素P450 1B1(CYP1B1)抑制剂的研究概况。CYP1B1是一类在导致癌变的雌激素代谢和激活芳香烃化合物致癌性中起重要作用的酶，在正常组织中通常不表达而在许多肿瘤组织中表达活跃，同时对许多抗癌药物的代谢具有重要影响。这表明CYP1B1抑制剂有望成为又一类肿瘤治疗药物。     

间质作用因子1通过正调控窖蛋白1抑制FBJ-S1-H的迁移性

目的证明间质作用因子(stromal interaction molecule1,Stim1)在FBJ诱导的小鼠骨肉瘤细胞中的抑癌作用。方法在Stim1高表达的FBJ-S1-H细胞采用Stim1以siRNA干扰技术得到Stim1沉默的几株S1-H单克隆细胞株,通过细胞行为学方法和RT-PCR技术对其mRNA进行研究,通过明胶酶谱法对细胞基质金属酶活性进行研究。结果通过细胞行为学方法证明,Stim1的沉默提高了细胞的迁移性,通过对mRNA表达的研究发现,Stim1沉默引起了多种基因表达的变化,其中包括基质金属酶9(matrix mexalloprotelnase 9,MMP-9)的升高,窖蛋白(caveolinl,Cav1),甾醇调控因子Srebf1的降低等,提高单克隆细胞中的Cav1含量可以使细胞迁移性降低。结论实验结果证明在FBJ-S1-H细胞中,Stim1能够抑制细胞的移动性,沉默Stim1的表达能够提高细胞的迁移性。     

内皮素-1与内皮素-1受体拮抗剂

内皮素－1（ET－1）是一种内源性的具有强缩血管活性的肽类物质，参与多种心血管疾病的病理生理过程。ET－1经与ET－1受体结合而发挥作用，多种不同结构类型的ET－1受体拮抗剂对某些心血管疾病均有良好的疗效。本文综述了近年来在ET－1及其受体拮抗剂等方面的研究进展。     

Pharmacological characterization of A-131701, a novel α1-adrenoceptor antagonist selective for α1A- and α1D-compared to α1B-adrenoceptors

New compounds selective for α_1A-adrenoceptors in the prostate may offer enhanced efficacy for benign prostatic hyperplasia (BPH), with fewer side effects than current treatment. A-131701 (3-[2-((3aR,9bR)-cis-6-methoxy-2,3,3a,4,5,9b,hexahydro-[1H]-benz[e]isoindol-2-yl)ethyl]pyrido[3′,4′:4,5]thieno [3,2-d]pyrimidine-2,4(1H,3H)-dione), from a novel class of benz[e]isolindole pyridothienopyrimidines and pyridothienopyrazines, is selective for α_1a- and α_1d-adrenoceptors in radioligand binding studies (0.22 nM at α_1a-, 0.97 nM at α_1d-) compared to α_1b-sites (2.5 nM) and in isolated tissue bioassays (pA₂ values of 8.9–9.0 for α_1A-receptors in rat vas deferens or canine prostate strips, 9.1 at α_1D-sites (rat aorta)), compared to 7.9 at α_1B-sites (rat spleen). A-131701 also potently blocked radioligand binding to α₁-adrenoceptors in canine and human prostatic membranes, but was considerably weaker at α₂-adrenoceptors. In isoflurane-anesthetized dogs, A-131701 antagonized epinephrine-induced increases in intraurethral pressure (IUP) with a pseudo-pA₂ value of 8.17. In spontaneously hypertensive rats, A-131701 caused transient decreases in mean arterial blood pressure (MABP) and transient tachycardia. The area under the curve (AUC₀→₆₀ min) for the hypotensive response was dose-related, with a log index value for A-131701 of 5.33, suggesting a selectivity of >600-fold comparing IUP to MABP effects. In pentobarbital-anesthetized dogs, A-131701 was more potent in blocking phenylephrine (PHE)-induced increases in IUP (pseudo-pA₂ = 8.0) compared to concurrently measured MABP (pseudo-pA₂ = 7.2), or sixfold selective. Doses greater than 1,000 nmol/kg i.v. of A-131701 were required to lower blood pressure by 10 mm Hg in these dogs (pED₁₀ =. 5.57), indicating a uroselectivity ratio of >250, superior to doxazosin, terazosin, or tamsulosin. Thus, A-131701 is selective for α_1A- and α_1D- vs. α_1B-adrenoceptors in vitro, and prostatic function vs. blood pressure effects in vivo, which may provide therapeutic advantages in the treatment of BPH. Drug Dev. Res. 44:140–162, 1998. © 1998 Wiley-Liss, Inc.