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Pharmacological Action and Molecular Mechanism of formononetin

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Asian Agricultural Research 2023年第11期15卷 35-37,41页

作者： Xichun HUANG Wenshuang HOU Quan QUAN Jingchao WANG Mingxinzhi WANG Xinyu LIU Chenghao JIN College of Life Science and Biotechnology Heilongjiang Bayi Agricultural UniversityDaqing 163319China

formononetin is an isoflavone phytoestrogen mainly existing in leguminous plants such as Trifolium pretense,Spatholobus suberectus,*** have proved that formononetin has good anti-inflammatory,antitumor,antioxidant,antidepressant,anxiolytic and other pharmacological *** paper summarizes the pharmacological action and molecular mechanism of formononetin,in order to provide a theoretical basis for the development and clinical application of formononetin.

关键词： formononetin Anti-inflammatory Antitumor Antioxidant Antidepressant Anxiolytic

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艾迪注射液药物相互作用研究

艾迪注射液药物相互作用研究

作者：徐洁南京中医药大学

学位级别：硕士

第一章液相串联质谱同时测定艾迪注射液中9种成分的含量目的：建立LC-MS方法,同时测定艾迪注射液中斑蝥素,人参皂苷Rg1、Re、 Rb1,异秦皮啶,紫丁香苷,芒柄花素,黄芪甲苷及绿原酸9种成分。方法：使用Waters Quattro Micro液质联用仪,色... 详细信息

第一章液相串联质谱同时测定艾迪注射液中9种成分的含量目的：建立LC-MS方法,同时测定艾迪注射液中斑蝥素,人参皂苷Rg1、Re、 Rb1,异秦皮啶,紫丁香苷,芒柄花素,黄芪甲苷及绿原酸9种成分。方法：使用Waters Quattro Micro液质联用仪,色谱柱为Agilent Zorbax SB C18(100mm×3.0mm,3.5μm);流动相为乙腈-0.1%甲酸,梯度洗脱;柱温23℃;体积流量0.22mL/min。结果：4个批次艾迪注射液中各成分平均质量浓度分别为(1.88±0.03)、(0.73±0.12)、(0.66±0.28)、(3.13±0.66)、(7.77±0.56)、(21.71±0.64)、(0.46±0.07)、(17.44±1.00)、(108.76±5.86)μg/mL。结论：建立的LC-MS方法,操作简单、快速、灵敏度高,可用于艾迪注射液的质量控制。第二章艾迪注射液对大鼠肝S9蛋白与细胞色素P450含量的影响目的：研究艾迪注射液对大鼠肝S9蛋白与细胞色素P450含量的影响。方法：将20只SD大鼠随机平均分为空白对照组(生理盐水10mL·kg-1)、阳性对照组(苯巴比妥钠80mg·kg-1)、艾迪注射液正常剂量组(10mL·kg-1)、高剂量组(20mL·kg-1)。各组腹腔注射连续给药10天,制备大鼠肝S9。采用Lowry法测定蛋白浓度;CO还原差示光谱法测定肝S9细胞色素P450酶含量。实验数据用单因素方差分析进行统计。结果：艾迪注射液正常剂量组和高剂量组均能提高大鼠肝重指数和肝S9细胞色素P450含量,但均低于苯巴比妥钠组。两个剂量组之间的差异不显著(P>0.05)。结论：艾迪注射液对大鼠肝S9中细胞色素P450可能有诱导作用,且不呈剂量依赖性。第三章Cocktail法评价艾迪注射液对大鼠体内P450酶活性的影响目的：采用Cocktail探针药物法评价艾迪注射液对大鼠体内6种CYP450亚型酶活性的影响,为临床联合用药时考察相互作用提供酶学依据。方法：分别选用甲苯磺丁脲,氯唑沙宗,茶碱,咪达唑仑,奥美拉唑及右美沙芬作为CYP2C9、 CYP2E1、CYP1A2、CYP3A4、CYP2C19、CYP2D6的探针底物。大鼠腹腔注射艾迪注射液10ml·kg-1,连续给药10天。采用LC-MS测定给药前后大鼠体内6种混合探针的血药浓度,计算药动学参数。结果：艾迪注射液连续给药10d后,甲苯磺丁脲和氯唑沙宗的AUC0-24h与给药前相比,有降低趋势(P0.05)。结论：艾迪注射液对大鼠体内CYP2C9、CYP2E1酶有诱导作用;对CYP2C19酶有弱的抑制作用,对CYP2D6酶有中强抑制作用;而对CYP1A2和CYP3A4酶的活性无显著影响。第四章艾迪注射液合用昂丹司琼对大鼠药动学的影响目的建立大鼠体内昂丹司琼血药浓度的]HPLC检测方法,考察联合应用艾迪注射液对昂丹司琼血药浓度的影响。方法大鼠腹腔注射艾迪注射液10ml·kg-1,连续给药10天。在给予艾迪注射液前后灌胃给予昂丹司琼4mg·kg-1,在不同时间取血,血浆经乙酸乙酯处理后,进HPLC分析,计算血药浓度。实验前后两轮自身对照,比较药动学参数的变化。结果联合应用艾迪注射液后,昂丹司琼的Cmax和AUC((0-t)显著性降低(P<0.01),代谢加快,AUC((0-t)是给药前的0.58倍。结论临床两者合并用药时,需加大昂丹司琼用药量,保证临床疗效。

关键词： Aidi injection cantharidin ginsenoside Rg1 ginsenoside Re ginsenoside Rb1 Isofraxidin Syringin formononetin Astragaloside chlorogenicacid LC/MS Cocktail probe drugs LC-MS/MS CYP450 ondansetron interaction HPLC

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脑红蛋白的表达调控和功能研究

脑红蛋白的表达调控和功能研究

作者：王婷婷湖南师范大学

学位级别：硕士

脑红蛋白是内源性的神经保护分子,能够保护大脑神经元免受缺血缺氧、氧化应激以及β-淀粉样蛋白诱发的神经损伤。中风和神经退行性疾病能够引发内源的神经保护机制,对这一机制加以利用,可能引导治疗这两种疾病的新的方向。因此,能够上... 详细信息

脑红蛋白是内源性的神经保护分子,能够保护大脑神经元免受缺血缺氧、氧化应激以及β-淀粉样蛋白诱发的神经损伤。中风和神经退行性疾病能够引发内源的神经保护机制,对这一机制加以利用,可能引导治疗这两种疾病的新的方向。因此,能够上调大脑中内源性脑红蛋白的药物在预防和治疗中风以及其他神经系统疾病方面具有很大的潜力。近年的研究发现,丙戊酸能够拮抗多种因素诱导的神经元损伤,具有保护大脑免受中风以及相关神经系统疾病诱发的神经损伤的能力。formononetin能够保护大脑神经元免受L-谷氨酸或β-淀粉样蛋白诱导的氧化应激和神经毒性。这些结果说明,丙戊酸和formononetin可能作为神经保护类药物。但是,丙戊酸和formononetin的神经保护作用机制还有待进一步研究。我们通过Western blot、实时定量PCR和荧光素酶活性分析等实验研究发现,丙戊酸和formononetin能够上调人和小鼠脑红蛋白的蛋白水平和mRNA水平,并能够上调脑红蛋白基因的启动子活性;转录因子CREB在丙戊酸和formononetin诱导的脑红蛋白启动子活性过程中起介导作用;CREB特异性抑制剂KG-501和CREB显性负突变体KCREB均对丙戊酸和formononetin诱导的脑红蛋白基因的启动子活性有抑制作用;另外,MTT实验结果说明丙戊酸可能保护N2a神经瘤母细胞免受双氧水诱导的损伤,CRE2位点介导了 formononetin诱导的脑红蛋白基因启动子活性上调。简而言之,我们的研究数据表明,丙戊酸和formononetin通过CREB的介导诱导人和小鼠脑红蛋白的表达上调,进而发挥其神经保护作用。

关键词：脑红蛋白丙戊酸 formononetin CREB 神经保护机制

Molecular Mechanism of Induction on Apoptosis of Human Esophageal Cancer HCE-4 Cells by Active Components from Astragalus membranaceus

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Medicinal Plant 2018年第1期9卷 63-66页

作者： Jiaru WANG Yinghua LUO Xianji PIAO Chang LIU Yi ZHANG Hao WANG Jinqian LI Wanting XU Yang LIU Yiqin WU Chenghao JIN College of Life Science & Technology Heilongjiang Bayi Agricultural University College of Animal Science & Technology Heilongjiang Bayi Agricultural University Department of Gynaecology and Obstetrics The Fifth Affiliated Hospital of Harbin Medical University

[Objectives] To investigate the pharmacologic effects of active components from A. membranaceus on human esophageal cancer HCE-4 cells and its apoptosis mechanism. [Methods] The viabilities of HCE-4 cells were measured by MTT assay. The induction of active components from A. membranaceus on apoptosis of HCE-4 cells was detected by Annexin V-FITC/PI double staining. The apoptotic-related protein expression levels were determined by Western blotting. [Results] formononetin and astragaloside IV suppressed the proliferation of HCE-4 cells in a dose-dependent manner. The Annexin V-FITC/PI double staining results showed that formononetin and astragaloside IV could induce HCE-4 cells apoptosis in a time-dependent manner. The Western blotting results showed that formononetin and astragaloside IV could significantly down-regulate p-AKT,pro-caspase-3,and increase cle-caspase-3 protein expression in HCE-4 cells. [Conclusions]Active components from A. membranaceus such as formononetin and astragaloside IV significantly inhibited the proliferation of human esophageal cancer HCE-4 cells by inducing mitochondrial dependent apoptosis via AKT signaling pathway.

关键词： Human esophageal cancer HCE-4 cells formononetin Astragaloside IV Astragalus root extract Apoptosis AKT signal path way

Content Determination of Flavonoids in Radix Millettiae Speciosae from Different Areas of Guangxi

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Medicinal Plant 2019年第1期10卷 31-33页

作者： Jingwen GU Liuping WANG Dongping TU Xiajun ZHONG Guangxi International Zhuang Medicine Hospital Guangxi University of Chinese Medicine Guangxi Center for Zhuang and Yao Medicine Engineering and Technology Research

[Objectives] This study aimed to investigate the quality of Radix Millettiae Speciosae at different planting sites in Guangxi. [Methods] The content of flavonoids in Radix Millettiae Speciosae was determined by HPLC. The chromatographic conditions were as follows: column, ZORBAX SB-C_(18); mobile phase, acetonitrile-0.1% glacial acetic acid(42∶58); detection wavelength, 250(formononetin) and 310(maackiain) nm; flow rate, 1.0 mL/min; and column temperature, 30℃. [Results] Under the chromatographic conditions above, formononetin and maackiain could be completely separated from impurities. The standard curve had a good linear relationship(R^2>0.999 9). The precision and stability met analysis requirements. The average recovery rates were 97.53% and 98.54%, respectively, and the RSD values were 2.03% and 1.88%, respectively, indicating that the established method has good reproducibility, durability and accuracy. The content of formononetin was highest in the Radix Millettiae Speciosae from Yongning District, Nanning City, and the content of maackiain was highest in the Radix Millettiae Speciosae from Xichang Town, Hepu County. [Conclusions] The HPLC method established in this study is simple, accurate and stable. It can be used as a quality control method for Radix Millettiae Speciosae. The contents of flavonoids in Radix Millettiae Speciosae from Yongning District, Nanning City and Xichang Town, Hepu County were higher than those from other planting bases.

关键词： Radix Millettiae Speciosae Flavonoids formononetin Maackiain HPLC