HPLC-Q-Exactive-Orbitrap-MS法鉴定森登-4味涂剂中化学成分

目的 采用高效液相色谱–四级杆/静电场轨道阱高分辨质谱（HPLC-Q-Exactive-Orbitrap-MS）法鉴定森登-4味涂剂中化学成分。方法 色谱条件：Shim-pack GIST-HP C18色谱柱（150 mm×4.6 mm,5μm）,流动相为乙腈–0.1%甲酸水溶液,梯度洗脱,体积流量0.35 mL/min,柱温为40℃,进样量为5μL。质谱条件：检测方式Full MS/dd-MS2,扫描范围m/z110～1 200。电喷雾离子源（ESI）,正、负离子模式扫描,喷雾电压4.20 kV（+）、3.80 kV（-）,碰撞能量（CE）30 eV。通过全方化学成分信息库,结合多级质谱碎片信息、对照品保留时间,鉴定森登-4味涂剂中化学成分。结果 在森登-4味涂剂中共鉴定出60个成分,包括16个黄酮类成分、13个有机酸类成分、11个环烯醚萜类成分、10个鞣质类成分、4个氨基酸类成分、4个脂肪酸类成分、2个三萜类成分,并考察了山栀苷、槲皮素、阿魏酸、克里拉京、齐墩果酸、脯氨酸、亚油酸的MS2图和可能的裂解途径。结论 对森登-4味涂剂中化学成分进行了快速、准确的鉴定,为其质量标准建立、药...     

旱莲草单体对人孕烷X受体调控CYP3A4转录表达的调节作用

目的 研究旱莲草5种中药单体木犀草素、芹菜索、槲皮素、刺囊酸和蟛蜞菊内酯是否可以通过PXR的激活而诱导CYP3A4转录表达.方法 采用MTT法检测5种中药单体对人结肠癌细胞系LSI74T细胞的毒性,采用脂质体瞬时转染的方法,在LSI74T细胞中同时转入人PXR表达质粒、CYP3A4报告质粒以及内参质粒,建立双荧光报告基因体系,与不同浓度及不同给药时间的中药单体共同孵育,检测细胞中荧光素酶.结果 5种中药单体对LS174T的IC50值分别为木犀草素(71.08±5.35) μmol/L、芹菜素(185.54±9.57)μmol/L、槲皮素(112.68±8.34) μmol/L、蟛蜞菊内酯(36.01±9.37) μmol/L、刺囊酸(26.04±5.62) μmol/L.刺囊酸(5、10和20μmol/L)分别诱导PXR调控的CYP3A4表达(1.52±0.31)倍、(1.95±0.31)倍和(2.57±0.23)倍,蟛蜞菊内酯(5、10和20 mol/L)分别诱导PXR调控的CYP3A4表达(1.57±0.30)倍、(1.91±0.25)倍和(2.14±0.51)倍,木犀草素、槲皮素、芹菜素在试验的浓度范围内不能明显诱导LS174T细胞中PXR调控的CYP3A4表达.在不同给药时间试验中,10 μmol/L和20μmol/L的刺囊酸在12 ～ 48 h能诱导PXR调控的CYP3A4表达,诱导能力随时间的延长而呈增强的趋势,各浓度的最大诱导倍数均在48 h出现;10 μmol/L和20μmol/L的蟛蜞菊内酯只能在12～24 h诱导PXR调控的CYP3A4表达,在48 h时2个浓度组均出现诱导表达倍数的下调.结论 刺囊酸和蟛蜞菊内酯在LS174T细胞系中可以通过激活PXR诱导CYP3A4转录表达,而木犀草素、芹菜素、槲皮素无此作用.     

高效液相色谱法测定金钱草颗粒中槲皮素含量

目的建立测定金钱草颗粒中槲皮素含量的高效液相色谱法。方法色谱柱为Phenomenex Prodigy ODS3 C18柱(250 mm×4.6 mm,5μm),流动相为甲醇-0.4%磷酸溶液(50∶50),流速为1.0 mL/min,检测波长为360 nm,柱温为30℃。结果槲皮素质量浓度在10.4～104.0μg/mL范围内与峰面积呈良好线性关系(r=0.999 9),平均回收率为99.92%,RSD=1.07%(n=6)。结论该方法简便、准确、重复性好,可用于测定金钱草颗粒的含量。     

槲皮素对淀粉样纤维细胞毒性的抑制作用

目的采用人红细胞为实验模型,探索槲皮素对溶菌酶淀粉样纤维细胞毒性的抑制作用。方法制备溶菌酶淀粉样纤维,在溶菌酶纤维溶液中加入槲皮素,用原子力显微镜观察槲皮素对淀粉样纤维的分解作用;在红细胞悬液中加入溶菌酶淀粉样纤维和槲皮素,用扫描电镜观察细胞形态;SDS凝胶电泳分离细胞膜蛋白,检测在槲皮素存在的条件下,溶菌酶纤维诱导膜蛋白聚集的作用。结果槲皮素能够破坏淀粉样纤维结构,使纤维解聚,从而使溶菌酶淀粉样纤维的细胞损害作用降低,包括抑制溶菌酶纤维诱导的细胞聚集和细胞膜蛋白交联。结论槲皮素能够破坏成熟的溶菌酶淀粉样纤维结构,抑制淀粉样纤维对细胞膜的损害作用。槲皮素的这种作用与其分子的疏水性和抗氧化作用有关。     

甘肃省不同产地萱草花蕾中芦丁、槲皮素、山柰酚的测定

目的:建立HPLC法同时测定萱草花蕾中芦丁、槲皮素、山柰酚3个黄酮类化合物的含量,并比较不同产地萱草中这3个化合物的含量。方法:采用Agilent SB-C18(4.6 mm×250 mm,5μm)色谱柱,以甲醇(A)-pH 3磷酸水溶液(B)为流动相,梯度洗脱(0～10 min,20%A;10～10.5 min,20%A→45%A;10.5～27.5 min,45%A;27.5～28 min,45%A→60%A;28～45min,60%A→20%A),流速0.7 mL.min-1,检测波长360 nm,柱温33℃,进样量20μL。结果:芦丁、槲皮素及山柰酚浓度分别在32.6～326μg.mL-1(r=0.9999)、1.06～10.6μg.mL-1(r=0.9997)和0.32～3.2μg.mL-1(r=0.9997)范围内线性关系良好;平均加样回收率(n=6)分别为99.53%(RSD=1.8%),96.96%(RSD=1.7%),98.75%(RSD=1.9%)。结论:该方法简便,操作简单,结果准确,重复性好,为评价萱草药材的质量提供了可靠的分析方法,可作为萱草中黄酮类化合物的含量测定方法。     

不同产地胡芦巴中总黄酮和槲皮素的含量测定

目的测定胡芦巴 (Trigonellafoenum graecumL )种子中总黄酮及槲皮素的含量。 方法总黄酮含量测定 :以芦丁为对照品 ,采用分光光度法 ,于 5 10nm处测定 ;槲皮素含量测定 :采用高效液相色谱法 ,色谱柱为KromasilODS— 1,流动相为乙腈 1g·L-1磷酸溶液 (体积比 30∶70 ) ,检测波长 36 0nm。结果总黄酮在 8 4 8～ 5 0 88mg·L-1(r =0 9996 )、槲皮素在 0 18～ 3 6 8mg·L-1(r =0 9995 )内线性关系良好 ;总黄酮和槲皮素的平均回收率分别为 99 4 %、99 3%。结论建立的方法可为胡芦巴的质量控制提供依据     

高效液相色谱法测定金钱草胶囊中槲皮素的血药浓度

目的建立金钱草胶囊中槲皮素的血药浓度测定方法,为临床应用和药代动力学研究提供技术参考。方法采用高效液相色谱-紫外检测方法测定,色谱柱为Kromasil C8柱(250 mm×4.6 mm,5μm),流动相为甲醇-乙腈-0.4%磷酸溶液(40∶10∶50),流速1 mL/min,柱温30℃,进样量20μL,检测波长360 nm。血样提取方法,以β-葡萄糖苷酸酶水解后,以乙酸乙酯提取,氮气流吹干后复溶进样。结果槲皮素保留时间约为8.8 min,金钱草胶囊中槲皮素的血药浓度线性范围0.08～12μg/mL,最低定量限为0.05μg/mL,检测限为0.02μg/mL。方法平均回收率为97.50%～102.50%,水解提取回收率为76.46%～87.43%,日内精密度RSD为4.18%～6.17%,日间精密度RSD为5.08%～8.63%。结论方法简便,分析速度快,灵敏度较高,重复性好,适用于临床血药浓度测定及药代动力学研究。     

Phosphatidylinositide 3-kinase inhibition: A new potential target for the treatment of polycystic ovarian syndrome

Context: Quercetin, a flavonoid, has been tried in traditional medicine for treating many disorders and reported to have inhibitory action on PI3 kinase.

Objective: This study investigates the effect of quercetin on testosterone propionate induced polycystic ovary syndrome (PCOS) model, which shows both metabolic and endocrine features of PCOS.

Materials and methods: Female pre-pubertal Sprague–Dawley rats were randomly divided into four groups: normal control, PCOS control, quercetin, and metformin treated. PCOS was induced by testosterone propionate (10?mg/kg, s.c.) and treatments were carried out orally at the dose of 150?mg/kg from the 6th week. At the 6th and 10th week, blood was collected to investigate metabolic indices, and reproductive biochemical parameters including morphology of ovary, uterus, and estrous cyclicity were assessed. The ovaries were processed to determine CYP17A1 gene expression.

Results: The treatment with quercetin did not modify body weight gain but uterine (296.7?±?5.11 versus 263.0?±?8.60?mg) and ovary weights (49.5?±?1.93 versus 37.8?±?3.43?mg) were found to be decreased significantly (p <0.05) as compared with the PCOS control group. The PCOS control group showed hyperinsulinemia, hyperandrogenemia, and dyslipidemia. Treatment with quercetin showed statistically significant (p?<0.01) improvement in insulin (12.46?±?0.3 versus 10.0?±?0.28 μU/ml), testosterone (0.65?±?0.02 versus 0.29?±?0.02 μU/ml), luteinising hormone (20.6?±?0.28 versus 15.1?±?0.36 U/ml), and lipid profile. Histological examination of ovary and uterus confirmed the disease occurrence and remission state in the diseased and treated groups, respectively. Quercetin also demonstrated PI3 kinase inhibition in a docking study and decreased CYP17A1 gene expression.

Discussion and conclusion: Thus, we can conclude that quercetin may have beneficial effect in PCOS by virtue of inhibition of PI3K which attributes to a decrease in the expression of CYP17A1 gene, having a key role in steroidogenesis.     

Metabonomic analysis of quercetin against the toxicity of chronic exposure to a mixture of four organophosphate pesticides in rat plasma

1.?A metabonomics approach was performed to investigate the effect of quercetin on the toxicity of chronic exposure to a mixture of four organophosphate pesticides (OPs) at their corresponding no-observed-adverse-effect level (NOAEL). The rats were divided into six groups (n?=?10/group): control, two different doses of quercetin, OPs mixture and different doses of quercetin plus OPs mixture-treated groups.

2.?Nine metabolites, including two quercetin metabolites and seven endogenous metabolites were identified in plasma. The intensities of metabolites significantly changed in the OP mixture-treated group compared with the control group (p?<?0.01), such as lysoPE (16:0/0:0), lysoPC (17:0/0:0), lysoPC (15:0/0:0) and 4-pyridoxic acid, significantly increased; by contrast, the intensities of arachidonic acid and citric acid significantly decreased. Anomalous intensity changes in aforementioned metabolites were alleviated in the OP mixture plus 50?mg/kg?bw/d quercetin-treated group compared with the OP mixture-treated group (p?<?0.05).

3.?The results indicated that quercetin elicited partial protective effects against the toxicity induced by a mixture of OPs, which include regulation of lipid metabolism, improvement of tricarboxylic acid (TCA) cycle disorders, enhancement of antioxidant defence system to protect the liver.     

Quercetin-loaded poly (lactic-co-glycolic acid)-d-α-tocopheryl polyethylene glycol 1000 succinate nanoparticles for the targeted treatment of liver cancer

Utilization of quercetin (QT) in clinics is limited by its instability and poor solubility. To overcome these disadvantages, we prepared QT as QT-loaded PLGA-TPGS nanoparticles (QPTN) and examined its properties and therapeutic efficacy for liver cancer. QT-loaded PLGA nanoparticles (QPN) and QT/coumarin-6-loaded PLGA-TPGS nanoparticles (QCPTN) with coumarin-6 as a fluorescent marker were also prepared to investigate the cellular uptake by HepG2 and HCa-F cells using a confocal laser scanning microscope (CLSM), and their effects on apoptosis of HepG2 cells were assessed with flow cytometry. The results measured using transmission electron microscopy, scanning electron microscopy and size analyses indicated that QPTN were stably dispersed sphere with diameter in the range of 100-200?nm. It indicated that the QT loading and encapsulation efficiency in QPTN reached 21.63% and 93.74%, respectively, and the accumulative drug release of QPTN was 85.8%, the QCPTN uptake in HCa-F and HepG2 cells were 50.87% and 61.09% using HPLC analysis, respectively. The results determined using an Annexin-PI flow cytometry indicated that QPTN could induce HepG2 cell apoptosis in a dose dependent manner. The results of histological examination and HPLC analysis confirmed that QPTN was targeted to liver cells. In vivo analysis using solid tumor-bearing mouse model indicated that QPTN could suppress tumor growth by 59.07%. Moreover, all the studied properties of QPTN were more desirable than those of QT-loaded PLGA nanoparticles (QPN). In conclusion, QPTN could be used as a potential intravenous dosage form for the treatment of liver cancer owing to the enhanced pharmacological effects of QT with increased liver targeting.