RP-HPLC法测定三七丹胶囊中人参皂苷Rb1、人参皂苷Rg1和三七皂苷R1的含量

目的：建立反相高效液相色谱法测定三七丹胶囊中人参皂苷Rb1、人参皂苷Rg1和三七皂苷R1的含量测定方法。方法：采用Hibar ODS C18（4.6mm×250mm,5μm）色谱柱,流动相为乙腈-水,梯度洗脱,流速1.0ml·min-1,柱温30℃,检测波长203nm。结果：人参皂苷Rb1、人参皂苷Rg1及三七皂苷R1分别在0.5246～5.2464μg（r=0.9997）、0.6792～6.7920μg（r=0.9998）和0.2542～2.5424μg（r=0.9997）范围内线性关系良好,平均回收率分别为人参皂苷Rb199.12%（RSD=0.61%,n=6）、人参皂苷Rg197.50%（RSD=1.63%,n=6）、三七皂苷R197.43%（RSD=1.04%,n=6）。结论：该方法定量简单、准确、重复性好,可作为三七丹胶囊的质量控制方法。     

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。     

血塞通片中三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1的含量测定

目的建立测定血塞通片中三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1含量的反相高效液相色谱（RP-HPLC）法。方法色谱柱为DiamonsilC18柱（250mm×4.6mm,5μm）,柱温30℃,采用乙腈-水线性梯度洗脱,检测波长203nm,流速1.0mL/min。结果样品中的3种皂苷分离良好,三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1进样量分别在2.555～12.775μg（r=0.9996）,8.115～40.575μg（r=0.9999）和7.665～38.325μg（r=0.9998）范围内与峰面积线性关系良好,平均回收率分别为99.33%,99.54%,99.82%。结论所用RP-HPLC法操作简便、结果准确,重现性好,可用于血塞通片的三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1含量测定。     

RP-HPLC法同时测定丹七片中三七皂苷R_1和人参皂苷Rg_1的含量

目的:建立以反相高效液相色谱法同时测定丹七片中三七皂苷R1和人参皂苷Rg1含量的方法。方法:色谱柱为KromasilC18(250mm×4.6mm,5μm),流动相为乙腈-水-磷酸(20.5∶79.5∶0.02),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、人参皂苷Rg1的检测浓度分别在4.67~46.68(r=0.999 0)、4.62~115.50μg.mL-1(r=0.999 9)范围内与各自峰面积积分值呈良好线性关系;平均回收率分别为97.93%和97.98%,RSD分别为1.31%(n=6)和1.38%(n=6)。结论:本方法简便、快速、准确,可用于丹七片的质量控制。     

HPLC法测定含三七类保健食品中三七皂苷R1、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1含量

目的 建立HPLC法测定保健食品中三七皂苷R1、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1含量的方法.方法 Waters Symmetry ShieldTM C18(250mm×4.6mm,5μm)色谱柱;柱温:30℃;流动相为乙腈-水,梯度洗脱;检测波长为203nm.结果 三七皂苷R1、人参皂苷Rg1、Re和Rb1分别在6.322~151.716μg·mL-1(r=0.9996)、25.043~601.029μg·mL-1(r=0.9999)、6.292~151.008μg·mL-1(r=0.9991)和25.018~600.430μg·mL-1(r=0.9999)范围内的线性关系良好;平均加样回收率分别为99.53%(RSD=1.18%,n=9),99.70%(RSD=1.34%,n=9),100.38%(RSD=1.15%,n=9)和99.94%(RSD=1.92%,n=9).结论 本法简便、准确、重现性好,可作为含三七类保健食品的质量控制方法.     

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。     

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。     

HPLC法测定云南白药胶囊中三七皂苷R_1和人参皂苷Rg_1的含量

目的采用高效液相色谱法测定云南白药胶囊中三七皂苷R1和人参皂苷Rg1含量。方法色谱条件:Phenomenex C18柱(100mm×4.6mm,2.6μm);流动相:乙腈-水(19∶81);流速:1mL.min-1;柱温:30℃;检测波长:203nm。结果三七皂苷R1和人参皂苷Rg1的线性范围分别为0.53～3.17μg(r=0.999 8),2.27～13.62μg(r=0.999 8);回收率(n=6)分别为100.6%(RSD=2.1%),97.7%(RSD=1.8%)。结论该方法准确可靠,重复性好,可用于测定云南白药胶囊中三七皂苷R1和人参皂苷Rg1含量。     

HPLC法同时测定野三七中三七皂苷R1及人参皂苷Rg1、Rb1、Rd的含量

目的：建立同时测定野三七中三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1及人参皂苷Rd四种三萜皂苷成分含量的高效液相色谱方法。方法：采用Agilent Zorbax SB-C18（250 mm×4.6 mm,5μm）色谱柱,流动相为乙腈（A）-水（B）,梯度洗脱（0～5 min,15%A→20%A,5～30 min,20%A→23%A,30～50 min,23%A→40%A,50～55 min,40%A→40%A）,流速1 ml·min-1,检测波长203 nm,柱温30℃。结果：三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1及人参皂苷Rd分别在0.122～2.440μg、0.524～10.480μg、0.454～9.080μg及0.370～7.400μg范围内呈良好线性关系（r〉0.999 7）;野三七药材中4种三萜皂苷成分的平均回收率（n=6）分别为98.83%、99.28%、100.82%、99.06%;RSD分别为1.34%、1.05%、1.14%、1.53%。结论：该方法简便、准确,分离效果好,首次用于野三七药材的质量评价。     

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。     

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。     

Effect of TCDD exposure on CYP1A1 and CYP1B1 expression in explant cultures of human endometrium.

Endometriosis is a debilitating disease estimated to affect 10% of reproductive-age women and characterized by the growth of endometrial tissue outside of the uterus. The present study characterizes a human endometrial explant culture model for studying the direct effects of TCDD exposure by assessing the expression of CYP1A1 and CYP1B1 mRNA (Northern blotting), protein (Western blotting), and activity (7-ethoxyresorufin-O-deethylase; EROD) in explants cultured with and without TCDD. Explants were obtained at laparoscopy or laparotomy from women undergoing surgery for tubal ligation, endometriosis, or pelvic pain unrelated to endometriosis. The explants were cultured with 10 nM estradiol (E(2)) or 1 nM E(2) plus 500 nM progesterone (P(4)) with or without TCDD (first 24 h). The expression of CYP1A1 and CYP1B1 mRNA was greatest with 10 nM TCDD and increased up to 72 h after initial exposure. EROD activity increased up to 120 h. Explants from a secretory phase biopsy became reorganized in culture and formed a new epithelial membrane, while maintaining basic endometrial morphology and viability for up to 120 h. At 24 h, TCDD significantly increased CYP1A1 and CYP1B1 mRNA, and at 72 h, TCDD significantly increased EROD activity and CYP1B1 protein compared to explants cultured without TCDD for similar times. CYP1B1 protein also exhibited substantial constitutive expression that was similar in uncultured biopsies, where CYP1B1 protein was immunolocalized in the cytoplasm of epithelial glands, with only occasional patches of protein in the surface epithelial membrane. In explants cultured with and without TCDD exposure, CYP1B1 protein was localized in the cytoplasm of the new surface epithelial membrane and glands closest to the surface. CYP1A1 protein was not detected in uncultured biopsies or explants. Both younger age (age 30 and under) and proliferative phase were associated with higher TCDD-induced EROD activity in specimens treated with E(2):P(4). No significant endometriosis-related differences were observed for any of the biomarkers, but the detection of disease-specific change was limited by small sample size and variability in tissue-cycle phase. The human endometrial explant culture model will be useful for future studies of the effects of dioxin-like compounds on human endometrium in relationship to cycle phase and hormonal exposure.