HTBA_LCMS_16114_MedChemExpress

Inhibitors, Agonists, Screening LibrariesSafety Data Sheet Revision Date:May-24-2017Print Date:May-24-20171. PRODUCT AND COMPANY IDENTIFICATION1.1 Product identifierProduct name :BLU-554Catalog No. :HY-100492CAS No. :1707289-21-11.2 Relevant identified uses of the substance or mixture and uses advised againstIdentified uses :Laboratory chemicals, manufacture of substances.1.3 Details of the supplier of the safety data sheetCompany:MedChemExpress USATel:609-228-6898Fax:609-228-5909E-mail:sales@1.4 Emergency telephone numberEmergency Phone #:609-228-68982. HAZARDS IDENTIFICATION2.1 Classification of the substance or mixtureNot a hazardous substance or mixture.2.2 GHS Label elements, including precautionary statementsNot a hazardous substance or mixture.2.3 Other hazardsNone.3. COMPOSITION/INFORMATION ON INGREDIENTS3.1 SubstancesSynonyms:NoneFormula:C24H24Cl2N4O4Molecular Weight:503.38CAS No. :1707289-21-14. FIRST AID MEASURES4.1 Description of first aid measuresEye contactRemove any contact lenses, locate eye-wash station, and flush eyes immediately with large amounts of water. Separate eyelids with fingers to ensure adequate flushing. Promptly call a physician.Skin contactRinse skin thoroughly with large amounts of water. Remove contaminated clothing and shoes and call a physician.InhalationImmediately relocate self or casualty to fresh air. If breathing is difficult, give cardiopulmonary resuscitation (CPR). Avoid mouth-to-mouth resuscitation.IngestionWash out mouth with water; Do NOT induce vomiting; call a physician.4.2 Most important symptoms and effects, both acute and delayedThe most important known symptoms and effects are described in the labelling (see section 2.2).4.3 Indication of any immediate medical attention and special treatment neededTreat symptomatically.5. FIRE FIGHTING MEASURES5.1 Extinguishing mediaSuitable extinguishing mediaUse water spray, dry chemical, foam, and carbon dioxide fire extinguisher.5.2 Special hazards arising from the substance or mixtureDuring combustion, may emit irritant fumes.5.3 Advice for firefightersWear self-contained breathing apparatus and protective clothing.6. ACCIDENTAL RELEASE MEASURES6.1 Personal precautions, protective equipment and emergency proceduresUse full personal protective equipment. Avoid breathing vapors, mist, dust or gas. Ensure adequate ventilation. Evacuate personnel to safe areas.Refer to protective measures listed in sections 8.6.2 Environmental precautionsTry to prevent further leakage or spillage. Keep the product away from drains or water courses.6.3 Methods and materials for containment and cleaning upAbsorb solutions with finely-powdered liquid-binding material (diatomite, universal binders); Decontaminate surfaces and equipment by scrubbing with alcohol; Dispose of contaminated material according to Section 13.7. HANDLING AND STORAGE7.1 Precautions for safe handlingAvoid inhalation, contact with eyes and skin. Avoid dust and aerosol formation. Use only in areas with appropriate exhaust ventilation.7.2 Conditions for safe storage, including any incompatibilitiesKeep container tightly sealed in cool, well-ventilated area. Keep away from direct sunlight and sources of ignition.Recommended storage temperature:Powder-20°C 3 years4°C 2 yearsIn solvent-80°C 6 months-20°C 1 monthShipping at room temperature if less than 2 weeks.7.3 Specific end use(s)No data available.8. EXPOSURE CONTROLS/PERSONAL PROTECTION8.1 Control parametersComponents with workplace control parametersThis product contains no substances with occupational exposure limit values.8.2 Exposure controlsEngineering controlsEnsure adequate ventilation. Provide accessible safety shower and eye wash station.Personal protective equipmentEye protection Safety goggles with side-shields.Hand protection Protective gloves.Skin and body protection Impervious clothing.Respiratory protection Suitable respirator.Environmental exposure controls Keep the product away from drains, water courses or the soil. Cleanspillages in a safe way as soon as possible.9. PHYSICAL AND CHEMICAL PROPERTIES9.1 Information on basic physical and chemical propertiesAppearance White to light yellow (Solid)Odor No data availableOdor threshold No data availablepH No data availableMelting/freezing point No data availableBoiling point/range No data availableFlash point No data availableEvaporation rate No data availableFlammability (solid, gas)No data availableUpper/lower flammability or explosive limits No data availableVapor pressure No data availableVapor density No data availableRelative density No data availableWater Solubility No data availablePartition coefficient No data availableAuto-ignition temperature No data availableDecomposition temperature No data availableViscosity No data availableExplosive properties No data availableOxidizing properties No data available9.2 Other safety informationNo data available.10. STABILITY AND REACTIVITY10.1 ReactivityNo data available.10.2 Chemical stabilityStable under recommended storage conditions.10.3 Possibility of hazardous reactionsNo data available.10.4 Conditions to avoidNo data available.10.5 Incompatible materialsStrong acids/alkalis, strong oxidising/reducing agents.10.6 Hazardous decomposition productsUnder fire conditions, may decompose and emit toxic fumes.Other decomposition products - no data available.11.TOXICOLOGICAL INFORMATION11.1 Information on toxicological effectsAcute toxicityClassified based on available data. For more details, see section 2Skin corrosion/irritationClassified based on available data. For more details, see section 2Serious eye damage/irritationClassified based on available data. For more details, see section 2Respiratory or skin sensitizationClassified based on available data. For more details, see section 2Germ cell mutagenicityClassified based on available data. For more details, see section 2CarcinogenicityIARC: No component of this product present at a level equal to or greater than 0.1% is identified as probable, possible or confirmed human carcinogen by IARC.ACGIH: No component of this product present at a level equal to or greater than 0.1% is identified as a potential or confirmed carcinogen by ACGIH.NTP: No component of this product present at a level equal to or greater than 0.1% is identified as a anticipated or confirmed carcinogen by NTP.OSHA: No component of this product present at a level equal to or greater than 0.1% is identified as a potential or confirmed carcinogen by OSHA.Reproductive toxicityClassified based on available data. For more details, see section 2Specific target organ toxicity - single exposureClassified based on available data. For more details, see section 2Specific target organ toxicity - repeated exposureClassified based on available data. For more details, see section 2Aspiration hazardClassified based on available data. For more details, see section 212. ECOLOGICAL INFORMATION12.1 ToxicityNo data available.12.2 Persistence and degradabilityNo data available.12.3 Bioaccumlative potentialNo data available.12.4 Mobility in soilNo data available.12.5 Results of PBT and vPvB assessmentPBT/vPvB assessment unavailable as chemical safety assessment not required or not conducted.12.6 Other adverse effectsNo data available.13. DISPOSAL CONSIDERATIONS13.1 Waste treatment methodsProductDispose substance in accordance with prevailing country, federal, state and local regulations.Contaminated packagingConduct recycling or disposal in accordance with prevailing country, federal, state and local regulations.14. TRANSPORT INFORMATIONDOT (US)This substance is considered to be non-hazardous for transport.IMDGThis substance is considered to be non-hazardous for transport.IATAThis substance is considered to be non-hazardous for transport.15. REGULATORY INFORMATIONSARA 302 Components:No chemicals in this material are subject to the reporting requirements of SARA Title III, Section 302.SARA 313 Components:This material does not contain any chemical components with known CAS numbers that exceed the threshold (De Minimis) reporting levels established by SARA Title III, Section 313.SARA 311/312 Hazards:No SARA Hazards.Massachusetts Right To Know Components:No components are subject to the Massachusetts Right to Know Act.Pennsylvania Right To Know Components:No components are subject to the Pennsylvania Right to Know Act.New Jersey Right To Know Components:No components are subject to the New Jersey Right to Know Act.California Prop. 65 Components:This product does not contain any chemicals known to State of California to cause cancer, birth defects, or anyother reproductive harm.16. OTHER INFORMATIONCopyright 2017 MedChemExpress. The above information is correct to the best of our present knowledge but does not purport to be all inclusive and should be used only as a guide. The product is for research use only and for experienced personnel. It must only be handled by suitably qualified experienced scientists in appropriately equipped and authorized facilities. The burden of safe use of this material rests entirely with the user. MedChemExpress disclaims all liability for any damage resulting from handling or from contact with this product.Caution: Product has not been fully validated for medical applications. For research use only.Tel: 609-228-6898 Fax: 609-228-5909 E-mail: tech@Address: 1 Deer Park Dr, Suite Q, Monmouth Junction, NJ 08852, USA。

西红花提取物调控免疫细胞，提高程序性死亡受体-1抑制剂治疗肺腺癌效果的实验研究作者：李诗颖李存雅张雪钟薏来源：《上海医药》2024年第01期摘要目的：多项研究提示，西红花提取物能影响肿瘤的发展进程。

本实验探究西红花提取物在肺腺癌小鼠模型中对肿瘤免疫微环境和免疫治疗的影响，为西红花提取物抗肿瘤研究提供更多基础性数据。

方法：构建Lewis肺癌细胞和萤光素酶稳定结合的小鼠皮下瘤模型，观察西红花提取物对小鼠皮下瘤和肿瘤免疫微环境的影响：运用活体成像技术跟踪肿瘤生长情况；运用流式细胞技术检测小鼠CD4+、CD8+ T细胞的数量及占比；运用反转录-聚合酶链式反应技术检测程序性死亡受体配体1、含有T细胞免疫球蛋白和黏蛋白结构域的蛋白3（T cell immunoglobulin and mucin domaincontaining protein 3， TIM3）、淋巴细胞活化基因-3（lymphocyte-activation gene-3， LAG3）、具有免疫球蛋白和ITIM结构域的T细胞免疫受体（T cell immunoreceptor with immunoglobulin and ITIM domain， TIGIT）、胸腺细胞选择相关的高迁移率族蛋白（thymocyte selection-associated high mobility group box， TOX）1、TOX2、TOX3基因的mRNA表达情况。

结果：与对照组相比，给予西红花提取物能一定程度地抑制小鼠皮下瘤的生长（P关键词西红花免疫微环境肺腺癌免疫治疗中图分类号：R965； R282.71 文献标志码：A 文章编号：1006-1533（2024）01-0003-09引用本文李诗颖，李存雅，张雪，等. 西红花提取物调控免疫细胞，提高程序性死亡受体-1抑制剂治疗肺腺癌效果的实验研究[J]. 上海医药， 2024， 45（1）： 3-11； 28.基金项目：上海市2022年度“科技创新行动计划”医学创新研究专项项目（22Y31920104）；上海市虹口区第二轮“国医强优”三年行动计划（2022—2024年）中西医结合重点专科、薄弱专科建设项目（HKGYQYXM-2022-10）；上海市2021年度“科技创新行动计划”扬帆计划项目（21YF444400）；上海市2022年度“科技创新行动计划”启明星培育（扬帆专项）项目（22YF1444900）；山东省乡村振兴基金会张秀兰慈善基金项目Experimental study of saffron extracts to modulate immune cells to improve the efficacy of a programmed death-1 inhibitor in the treatment of lung adenocarcinomaLI Shiying1， LI Cunya1， ZHANG Xue2， ZHONG Yi1（1. Department of Oncology， Shanghai TCM-Integrated Hospital， Shanghai University of Traditional Chinese Medicine， Shanghai 200082， China； 2. Shanghai Traditional Chinese Medicine Co.， Ltd.， Shanghai 200082， China）ABSTRACT Objective： A number of studies have shown that saffron extracts can affect the development of tumor. This study explored the effect of saffron extract on tumor immune microenvironment and immunotherapy in a mouse model of lung adenocarcinoma so as to provide more basic data for the anti-tumor research of saffron extracts. Methods： The transplanted tumor model of Lewis lung carcinoma-luciferase in mice was established to detect the effect of saffron extracts on the transplanted tumor in vivo. At the same time， the tumor growth was tracked by in vivo imaging technique. The number and proportion of CD4+ and CD8+ T cells were determined by flow cytometry. The mRNA levels of programmed death-ligand 1， T cell immunoglobulin and mucin domain-containing protein 3 （TIM3）， lymphocyte-activation gene-3 （LAG3）， T cell immunoreceptor with immunoglobulin and ITIM domain （TIGIT）， thymocyte selection-associated high mobility group box （TOX） 1， TOX2 and TOX3 were detected by reverse transcription-polymerase chain reaction （RT-PCR） and immunohistochemical techniques to verify the effect of saffron extracts on the regulation of tumor immune microenvironment. Results：Compared with the control group， the administration of saffron extracts could inhibit the growth of subcutaneous tumor in mice to a certain extent， and the number and proportion of CD4+ and CD8+ T cells were increased （PKEY WORDS saffron； immune microenvironment； lung adenocarcinoma； immunotherapy肿瘤是一类恶性疾病，2018年全球肿瘤死亡病例数达约960万人，较2008年增加26.3%，其中男性肿瘤死亡病例数增加最多的是肺癌，增加了23.4万人[1-2]。

Inhibitors, Agonists, Screening LibrariesSafety Data Sheet Revision Date:Nov.-23-2018Print Date:Nov.-23-20181. PRODUCT AND COMPANY IDENTIFICATION1.1 Product identifierProduct name :Gelucire 14/44Catalog No. :HY-Y1892CAS No. :121548-04-71.2 Relevant identified uses of the substance or mixture and uses advised againstIdentified uses :Laboratory chemicals, manufacture of substances.1.3 Details of the supplier of the safety data sheetCompany:MedChemExpress USATel:609-228-6898Fax:609-228-5909E-mail:sales@1.4 Emergency telephone numberEmergency Phone #:609-228-68982. HAZARDS IDENTIFICATION2.1 Classification of the substance or mixtureNot a hazardous substance or mixture.2.2 GHS Label elements, including precautionary statementsNot a hazardous substance or mixture.2.3 Other hazardsNone.3. COMPOSITION/INFORMATION ON INGREDIENTS3.1 SubstancesSynonyms:NoneFormula:N/AMolecular Weight:N/ACAS No. :121548-04-74. FIRST AID MEASURES4.1 Description of first aid measuresEye contactRemove any contact lenses, locate eye-wash station, and flush eyes immediately with large amounts of water. Separate eyelids with fingers to ensure adequate flushing. Promptly call a physician.Skin contactRinse skin thoroughly with large amounts of water. Remove contaminated clothing and shoes and call a physician.InhalationImmediately relocate self or casualty to fresh air. If breathing is difficult, give cardiopulmonary resuscitation (CPR). Avoid mouth-to-mouth resuscitation.IngestionWash out mouth with water; Do NOT induce vomiting; call a physician.4.2 Most important symptoms and effects, both acute and delayedThe most important known symptoms and effects are described in the labelling (see section 2.2).4.3 Indication of any immediate medical attention and special treatment neededTreat symptomatically.5. FIRE FIGHTING MEASURES5.1 Extinguishing mediaSuitable extinguishing mediaUse water spray, dry chemical, foam, and carbon dioxide fire extinguisher.5.2 Special hazards arising from the substance or mixtureDuring combustion, may emit irritant fumes.5.3 Advice for firefightersWear self-contained breathing apparatus and protective clothing.6. ACCIDENTAL RELEASE MEASURES6.1 Personal precautions, protective equipment and emergency proceduresUse full personal protective equipment. Avoid breathing vapors, mist, dust or gas. Ensure adequate ventilation. Evacuate personnel to safe areas.Refer to protective measures listed in sections 8.6.2 Environmental precautionsTry to prevent further leakage or spillage. Keep the product away from drains or water courses.6.3 Methods and materials for containment and cleaning upAbsorb solutions with finely-powdered liquid-binding material (diatomite, universal binders); Decontaminate surfaces and equipment by scrubbing with alcohol; Dispose of contaminated material according to Section 13.7. HANDLING AND STORAGE7.1 Precautions for safe handlingAvoid inhalation, contact with eyes and skin. Avoid dust and aerosol formation. Use only in areas with appropriate exhaust ventilation.7.2 Conditions for safe storage, including any incompatibilitiesKeep container tightly sealed in cool, well-ventilated area. Keep away from direct sunlight and sources of ignition.Recommended storage temperature:Pure form-20°C 3 years4°C 2 yearsIn solvent-80°C 6 months-20°C 1 monthShipping at room temperature if less than 2 weeks.7.3 Specific end use(s)No data available.8. EXPOSURE CONTROLS/PERSONAL PROTECTION8.1 Control parametersComponents with workplace control parametersThis product contains no substances with occupational exposure limit values.8.2 Exposure controlsEngineering controlsEnsure adequate ventilation. Provide accessible safety shower and eye wash station.Personal protective equipmentEye protection Safety goggles with side-shields.Hand protection Protective gloves.Skin and body protection Impervious clothing.Respiratory protection Suitable respirator.Environmental exposure controls Keep the product away from drains, water courses or the soil. Cleanspillages in a safe way as soon as possible.9. PHYSICAL AND CHEMICAL PROPERTIES9.1 Information on basic physical and chemical propertiesAppearance White to off-white (Oil)Odor No data availableOdor threshold No data availablepH No data availableMelting/freezing point No data availableBoiling point/range No data availableFlash point No data availableEvaporation rate No data availableFlammability (solid, gas)No data availableUpper/lower flammability or explosive limits No data availableVapor pressure No data availableVapor density No data availableRelative density No data availableWater Solubility No data availablePartition coefficient No data availableAuto-ignition temperature No data availableDecomposition temperature No data availableViscosity No data availableExplosive properties No data availableOxidizing properties No data available9.2 Other safety informationNo data available.10. STABILITY AND REACTIVITY10.1 ReactivityNo data available.10.2 Chemical stabilityStable under recommended storage conditions.10.3 Possibility of hazardous reactionsNo data available.10.4 Conditions to avoidNo data available.10.5 Incompatible materialsStrong acids/alkalis, strong oxidising/reducing agents.10.6 Hazardous decomposition productsUnder fire conditions, may decompose and emit toxic fumes.Other decomposition products - no data available.11.TOXICOLOGICAL INFORMATION11.1 Information on toxicological effectsAcute toxicityClassified based on available data. For more details, see section 2Skin corrosion/irritationClassified based on available data. For more details, see section 2Serious eye damage/irritationClassified based on available data. For more details, see section 2Respiratory or skin sensitizationClassified based on available data. For more details, see section 2Germ cell mutagenicityClassified based on available data. For more details, see section 2CarcinogenicityIARC: No component of this product present at a level equal to or greater than 0.1% is identified as probable, possible or confirmed human carcinogen by IARC.ACGIH: No component of this product present at a level equal to or greater than 0.1% is identified as a potential or confirmed carcinogen by ACGIH.NTP: No component of this product present at a level equal to or greater than 0.1% is identified as a anticipated or confirmed carcinogen by NTP.OSHA: No component of this product present at a level equal to or greater than 0.1% is identified as a potential or confirmed carcinogen by OSHA.Reproductive toxicityClassified based on available data. For more details, see section 2Specific target organ toxicity - single exposureClassified based on available data. For more details, see section 2Specific target organ toxicity - repeated exposureClassified based on available data. For more details, see section 2Aspiration hazardClassified based on available data. For more details, see section 212. ECOLOGICAL INFORMATION12.1 ToxicityNo data available.12.2 Persistence and degradabilityNo data available.12.3 Bioaccumlative potentialNo data available.12.4 Mobility in soilNo data available.12.5 Results of PBT and vPvB assessmentPBT/vPvB assessment unavailable as chemical safety assessment not required or not conducted.12.6 Other adverse effectsNo data available.13. DISPOSAL CONSIDERATIONS13.1 Waste treatment methodsProductDispose substance in accordance with prevailing country, federal, state and local regulations.Contaminated packagingConduct recycling or disposal in accordance with prevailing country, federal, state and local regulations.14. TRANSPORT INFORMATIONDOT (US)This substance is considered to be non-hazardous for transport.IMDGThis substance is considered to be non-hazardous for transport.IATAThis substance is considered to be non-hazardous for transport.15. REGULATORY INFORMATIONSARA 302 Components:No chemicals in this material are subject to the reporting requirements of SARA Title III, Section 302.SARA 313 Components:This material does not contain any chemical components with known CAS numbers that exceed the threshold (De Minimis) reporting levels established by SARA Title III, Section 313.SARA 311/312 Hazards:No SARA Hazards.Massachusetts Right To Know Components:No components are subject to the Massachusetts Right to Know Act.Pennsylvania Right To Know Components:No components are subject to the Pennsylvania Right to Know Act.New Jersey Right To Know Components:No components are subject to the New Jersey Right to Know Act.California Prop. 65 Components:This product does not contain any chemicals known to State of California to cause cancer, birth defects, or anyother reproductive harm.16. OTHER INFORMATIONCopyright 2018 MedChemExpress. The above information is correct to the best of our present knowledge but does not purport to be all inclusive and should be used only as a guide. The product is for research use only and for experienced personnel. It must only be handled by suitably qualified experienced scientists in appropriately equipped and authorized facilities. The burden of safe use of this material rests entirely with the user. MedChemExpress disclaims all liability for any damage resulting from handling or from contact with this product.Caution: Product has not been fully validated for medical applications. For research use only.Tel: 609-228-6898 Fax: 609-228-5909 E-mail: tech@Address: 1 Deer Park Dr, Suite Q, Monmouth Junction, NJ 08852, USA。

Inhibitors, Agonists, Screening Libraries Data SheetBIOLOGICAL ACTIVITY:AR–A014418 is a selective and effective GSK3β inhibitor with an IC 50 value of 104 nM, and has no significant inhibition on 26 other kinases.IC50 & Target: IC50: 104 nM (GSK3β)In Vitro: AR–A014418 inhibits tau phosphorylation at a GSK3–specific site (Ser–396) in 3T3 fibroblasts expressing human four–repeat tau protein with IC 50 of 2.7 μM, and protects cultured N2A cells from death induced by blocking PI3K/PKB pathway. In hippocampal slices, AR–A014418 inhibits neurodegeneration mediated by beta–amyloid peptide [1]. While in NGP and SH–5Y–SY cells,AR–A014418 reduces neuroendocrine markers and suppresses neuroblastoma cell growth [2].In Vivo: In ALS mouse model with the G93A mutant human SOD1, AR–A014418 (0–4 mg/kg, i.p.) delays the onset of symptoms,improves motor activity, slows down disease progression, and postpons the endpoint of the disease [3]. In addition, AR–A014418produces inhibition effect on acetic acid– and formalin–induced nociception in mice by modulating NMDA and metabotropic receptor signaling as well as TNF–α and IL–1β transmission in the spinal cord [4].PROTOCOL (Extracted from published papers and Only for reference)Kinase Assay:[1]The competition experiments are carried out in duplicate with 10 concentrations of the inhibitor inclear–bottomed microtiter plates. The biotinylated peptide substrate, biotin–AAEELDSRAGS(PO3H2)PQL, is added at a final concentration of 2 μM in an assay buffer containing 6 milliunits of recombinant human GSK3 (equal mix of both α and β), 12mM MOPS, pH 7.0, 0.3 mM EDTA, 0.01% β–mercaptoethanol, 0.004% Brij 35, 0.5% glycerol, and 0.5 μg of bovine serumalbumin/25 μL and preincubated for 10–15 min. The reaction is initiated by the addition of 0.04 μCi of [γ–33P]ATP and unlabeled ATP in 50 mM Mg(Ac)2 to a final concentration of 1 μM ATP and assay volume of 25 μL. Blank controls without peptide substrate are used.After incubation for 20 min at room temperature, each reaction is terminated by the addition of 25 μL of stop solution containing 5mM EDTA, 50 μM ATP, 0.1% Triton X–100, and 0.25 mg of streptavidin–coated SPA beads corresponding to appr 35 pmol of binding capacity. After 6 h the radioactivity is determined in a liquid scintillation counter. Inhibition curves are analyzed by non–linear regression using GraphPad Prism.Cell Assay: AR–A014418 is dissolved in DMSO.[1]Cell viability is assessed by calcein/propidium iodide uptake. Calcein AM is taken up and cleaved by esterases present within living cells, yielding yellowish–green fluorescence, whereas PI is only taken up by dead cells,which become orange–red fluorescent. In brief, N2A cells are cultured for 2 days in vitro and then treated with 50 μM LY–294002 in the presence of AR–A014418 or vehicle (DMSO) for 24 h. Subsequently, N2A cells are incubated for 30 min with 2 μM PI and 1 μM calcein–AM. The cultures are then rinsed three times with Hanks' buffered saline solution containing 2 mM CaCl 2, and the cells are visualized by fluorescence microscopy using a Zeiss Axiovert 135 microscope. Three fields (selected at random) are analyzed per well (appr 300 cells/field) in at least three different experiments. Cell death is expressed as percentage of PI–positive cells from the total number of cells. In every experiment, specific cell death is obtained after subtracting the number of dead cells present inProduct Name:AR–A014418Cat. No.:HY-10512CAS No.:487021-52-3Molecular Formula:C 12H 12N 4O 4S Molecular Weight:308.31Target:GSK–3; GSK–3Pathway:Stem Cell/Wnt; PI3K/Akt/mTOR Solubility:10 mM in DMSOvehicle–treated cultures.Animal Administration: AR–A014418 is formulated in normal saline.[3]First, to examine the effects of GSK–3 inhibition on the clinical symptoms, life span, and motor behavior function of ALS, 56 Tg mice are divided into four groups. In each group, 0.5 mL of normal saline is mixed with either 0 μg (control group), 1 μg (group A), 2 μg (group B) or 4 μg (group C) of AR–A014418 per gram of mouse, and injected intraperitoneally into 14 animals per group 5 days a week beginning 60 days after birth. The mice are sacrificed at the endpoint described below.References:[1]. Bhat R, Xue Y, Berg S, Structural insights and biological effects of glycogen synthase kinase 3–specific inhibitor AR–A014418. J Biol Chem. 2003 Nov 14; 278(46):45937–45.[2]. Carter YM, et al. Specific glycogen synthase kinase–3 inhibition reduces neuroendocrine markers and suppresses neuroblastoma cell growth. Cancer Biol Ther. 2014 May;15(5):510–5.[3]. Koh SH, et al. Inhibition of glycogen synthase kinase–3 suppresses the onset of symptoms and disease progression of G93A–SOD1 mouse model of ALS. Exp Neurol. 2007 Jun;205(2):336–46.[4]. Martins DF, et al. The antinociceptive effects of AR–A014418, a selective inhibitor of glycogen synthase kinase–3 beta, in mice. J Pain. 2011 Mar;12(3):315–22.Caution: Product has not been fully validated for medical applications. For research use only.Tel: 609-228-6898 Fax: 609-228-5909 E-mail: tech@Address: 1 Deer Park Dr, Suite Q, Monmouth Junction, NJ 08852, USA。

甲基百里香酚蓝络合剂-亚硝酸根络合吸附波的研究及应用第33卷2005年5月分析化学(FENXIHUAXUE)研究简报ChineseJournalofAnalyticalChemistry第5期675～679钴(Ⅱ).甲基百里香酚蓝络合剂.亚硝酸根络合吸附波的研究及应用杨丽珠2陈大茴刘传银2刘炜2陆光汉(温州医学院药学院,温州325027)(华中师范大学化学学院,武汉430079)摘要在pH8.5,0.04mol/LNH3?HO-NHC1缓冲溶液中,钴-甲基百里香酚蓝络合剂(MTB)在NaNO,存在下,于一1.23V(埘.SCE)产生一尖锐,灵敏的二次导数极谱波,峰电流与钴(Ⅱ)浓度在5.0×10～～2.0×10mol/L范围内呈线性关系;检出限为1.0×10～mol/L.研究了该波的性质及电极反应机理,证明该波为络合吸附波.峰电流由中心离子钴(Ⅱ)还原产生.络合物在汞电极上的饱和吸附量为5.18×10-9mol/cm,符合Frumkin等温式.测得吸附系数卢=6.86×10,自由能△=33.30kJ/tool;电子转移数,l=2.不可逆吸附的转移系数Ot=0.58,反应速率常数k.=2.26/s.方法用于VB.和模拟样中痕量钻的测定,结果满意.关键词钻,甲基百里香酚蓝络合剂,亚硝酸根,络合吸附波1引言钴是人体必需的微量元素,近年研究表明,钴与人体健康存在着密切的关系.心血管疾病与患者体内的微量元素钴长期缺乏有关,而治疗心血管疾病的常用中草药黄芪,玉竹等钴含量均较高.目前,测定钴的方法有分光光度法,原子吸收光谱法,ICP—AES,化学发光光度法,荧光分析法,等离子体质谱法及电化学分析方法等_6J.甲基百里香酚蓝络合剂用于Ca",Mg2离子的络合滴定分析早有报道,但用于极谱测钴离子的报道不多.本实验研究了该络合吸附波用于钴离子测定的最佳实验条件和电极反应机理,并用该方法测定了来VB:和模拟样中钴的含量.2实验部分2.1仪器与试剂JP-2,JP-303型示波极谱仪(成都仪器厂);三电极体系:滴汞电极为工作电极,饱和甘汞电极为参比电极,铂片为辅助电极;BASCV-50W电化学系统(美国BAS公司);883型笔录式记录极谱仪(上海分析仪器厂).甲基百里香酚蓝络合剂溶液:1.0×10mol/L;Co标准溶液:1000mg/L(北京冶金研究所),用时稀释至所需浓度;4.0mol/LNaNO2溶液(储于冰箱);O.2mol/L的NH3?H2O —NHC1缓冲溶液(pH8.5).所用试剂均为分析纯,实验用水为二次蒸馏水.2.2实验方法在10mL的比色管中,依次加人2mLNH,?H2O—NHC1缓冲液(pH8.5),4.00mol/LNaNO2溶液5.00mL,适量的Co溶液和5.0X10一mol/LMTB0.5mL,用水定容至10mL.摇匀静置30min后进行极谱测定,起始电位一0.60V,记录一1.23V处二次导数极谱波峰电流.3结果与讨论3.1最佳条件选择实验了NaAc—HAc,BR,PBS,六次甲基四胺等底液,只有在NH,?H:O—NHC1缓冲液中波形好,灵敏度高.反复实验表明,pH=8.5,NH,?H2O—NHC1浓度在0.04～0.075mol/L之间时,其峰的形状好,峰电流大且稳定.实验选择pH=8.5,0.04mol/LNH3?H2O—NHC1缓冲液.MTB浓度为2.0×102004-O3-O1收稿;2004-06-09接受676分析化学第33卷mol/L时,峰电流达最大值.随着NaNO浓度的增加,峰电流增加,当NaNO的浓度到达2.5mol/L峰电流最大,但稳定性不好,实验选择2.0mol/L的NaNO.实验表明,反应开始时,峰电流随时间的增加而增加.反应30min后基本稳定,且被测溶液的峰电流稳定时间在4h以上.3.2线性范围与检出限在上述选定条件下,二阶导数峰电流与Co浓度在5.0×10一-2.0×10mol/L之间呈线性关系,线性回归方程为i.(nA)=一110.53+36.21×10c(mol/L),相关系数r=0.9998,对空白溶液进行11次测定,根据3倍法公式3o'/K,求得检出限为1.0×10一mol/L.RSD为1.4%O3.3干扰物质的影响相对于5.0x10mol/LCon,控制误差在±10%以内干扰离子情况为:K,NH,Na可以大量存在,1000倍的C1一,NO;,sO:一,Ba不干扰(未达最大量),800倍的cu",zn不干扰,600倍的F一,400倍的Pb¨,Ag,100倍的ca",Al¨,4O倍的Mg",10倍的sn",Ni",2倍的ca¨,As.对测co不干扰,相同浓度的Fe¨,Fe干扰实验的测定,加入3×10-4mol/L的r可以掩蔽5倍的Fe离子.3.4络合物的组成和稳定常数的测定取CMTB+cc02+=5×10I¨mol/L,改变cMTB和Cco2+,用连续变换测得Co与MTB的组成比为1:1.同时固定C0n浓度,改变MTB浓度,根据文献[7]方法,当厅=1时直线的相关系数为,=0.9987,I1.=2或3时均为弯曲的曲线,故Co:MTB=1:1(图1).但Co的配位数为6,根据文献[8],NO;和底液中的NH;也参加了配位,生成一种混配活性络合物.再根据截距A=0.5512,=1.814×1OnA,斜率B=4.123×10-1.,利用公式1/.=1/一+1/(,oi[R]),求得=1.34×10.3.5富集电位和富集时间的影响实验发现,富集电位越正,峰电流越大,但富集电位正于一0.30V时,峰电流变化不大,峰形不太好,所以实验选择富集电位为一0.30V.峰电流随着富集时间的增长而增加,当浓度达到3.5×10mol/L时,峰电流达到极限,表明此时已到达吸附平衡.浓度愈大,达到吸附平衡的时间愈短.当富集时间达到90S时峰电流趋于平衡,峰电流不再增加.图2是络合物的循环伏安图,正扫时有一还原波出现,而反扫时无相应的氧化波,说明该波为不可逆波.连续扫描后峰电流逐渐下降.以上事实表明,该波具有吸附波的性质.3.6扫描速率的影响rCMTB)../10(mol/L)'.图1直线法测络合比Fig.1DeterminationofthecompositionofcomplexbymethodoflinepH:8.5,0.04mol/LNH3?H20?NH4C1+2.0md/LNaN02+1.0×10一tool/LCo2,a.1/ip—l/.lh,柚I(mls);b.1/i.一l/(.m).在5O～4oomV/s范围内改变扫描速率测定络合物峰电流和峰电位E.,发现峰电流随着扫速的增加而增大,由Randles—Sevcik公式可得:iP:2.69×10.DtJAc.以P—tJ作图,若为直线则说明反应物受扩散控制,若曲线上翘则说明反应物吸附.本实验所得曲线偏离直线向上弯曲.另外,实验发现,峰电位E随tJ增大而负移,E与lnv的关系为E=一1.289—0.022lnv,r=0.9998,再次证明了体系的不可逆的吸附性质.3.7直流极谱图在883型直流极谱仪上绘制极谱图,从图中可以看出,当溶液中不加co时或不加MTB时无峰出现;加入co"后,于一1.20V左右出现一个峰形极谱波,且波高随着co的浓度的增加而增大.改变汞柱高度测量其峰高的变化,发现峰高并不随汞柱高度的变化而变化.说明该波为典型的催化波].3.8吸附量和吸附等温线的测定记录不同浓度的络合物达到平衡时单扫伏安曲线.根据公式F=Q/nFA[1叫(悬汞电极的面积为第5期杨丽珠等:钴(Ⅱ)一甲基百里香酚蓝络合剂一亚硝酸根络合吸附波的研究及应用0.0163cm,F为法拉第常数),可求得不同浓度的平衡吸附量厂.实验发现,当c.>6.0×10mol/L以后,厂值基本恒定,说明已达到饱和吸附,通过计算饱和吸附量F.=5.18×10molJcm.则未饱和吸附时悬汞电极上覆盖度0=厂.根据该公式可计算不同浓度下的覆盖度0,作6}-c图,可得吸附等温曲线(图3曲线a).假定曲线a符合Langmuir等温式,0=/(1+),式中为吸附系数,c为吸附物的浓度.计算其等温曲线b,曲线b与a 偏离甚大,显然不是Langmuir型吸附.若符合Frumkin吸附等温式:o/(1一)=/3ce2~~(y为吸附因子),可以得曲线c,当c<3.5×10mol/L时曲线a和c符合较好.表明此条件下的吸附符合Frumkin等温式.求得吸附系数=6.86×10,吸附因子y=0. 229,由l=AG/RT得自由能AC,=33.30kJ/tool.由于值较大,说明吸附强烈.吸附因子为正,说明电极表面被吸附物有相互吸引作用¨.3.9电极反应电子数万,转移系数和电极反应速率常数.的测定二次导数极谱峰半峰宽28mV,根据公式=47.7/n【12],可求得n=1.70,该电极反应的电子转移数为2.由E.-lnv的直线关系斜率dE./dlnv=一RT/(鲫F)=一0.022(t=25oC)求得鲫=1.16,已测得//,=2,转移系数=0.58.确定循环伏安法扫速为50～350mV/s,作E.和Ep-lnv曲线,根据E.=E+(RI'/~nF)ln(R1'/anF)一(Rl'/anF)lnv¨引,作E.曲线,延长使口=0时,求得E=一1.223V,根据Ep-lnT)曲线的线性方程:E.=一1.289—0.0221nv(r=0.9998),可知当lnv 时,E.=E+(RT/o.nF)In(R鲫F).=一1.289V,代人斜率可求得表观反应速率常数.=2.26/s.3.10电极反应机理讨论为了研究电极反应机理,进行恒电位电解:按实验方法配制成5.0×10mol/LC02溶液40rnL,用大面积汞电极作阴极,在一1.35V处电解6h后,测得络合物的峰电流降低.将此溶液分图2络合物的循环伏安图Fig.2Cyclicvo]tammogramsofcomplexa.2.5×10～mol/LNTB0.04mol/LNH3?H20一NH4C1(pH=8.5)+2.0mol/LNaN02;b.a+1.0×10一'mol/LCo2':c.a+5.0×10'mol/LCo':d.a+1.0×10mol/LCo".富集(enrich)t=90s,静止(stand)t=108,口=101)mV/s.图3吸附等温曲线Fig.3Ad~rpdonisothermClll~e8a.实验值(experimentalcurve);b.I.angmmr公式计算值(calculatedfromequtionI.ang-mir);c.Fnm~kin公式计算值(calcI1laled￡romequtlonFnJmkln)o成3份:其中1份加入MTB溶液后,峰电流不变,第2份加入亚硝酸根溶液峰电流也不变,第3份加人Co峰电流增加,由此证明峰电流是由络合物的中心离子Co还原产生.而且,汞电极表面无氢气逸出,说明极谱波不是氢催化波.结合直流极谱的峰形,电毛细管曲线降低等实验事实,认为该极谱波为一吸附性的催化波.NO;的存在催化了电极反应,综合以上实验,可推测电极过程如下:Co(Ⅱ)+MTB—[CO(Ⅱ)一MTB][Co(U)一MTB]寻一[Co(U)]一MTB]8d[Co(U)一MTB]d+2e—}Co(0)+.MTB 1................................................................一JNo;3.11样品分析3.11.1维生素Bn中钴的测定取一支维生素Bl2针剂溶液(0.5mg/支,武汉制药厂,批号030816)于烧杯中,加入混合酸(HNO,:HCIO=3:1)5mL,加热分解其中的有机物,蒸发至近干,稍冷,重复2～3次,加入l0滴H0,蒸发至接近无色,加少量水并用NaOtt溶液调节至中性,加水定容至100mL.移取一定量上述试液,按实验方法配制溶液,进行极谱测定.结果见表1.与原子吸收光谱法比较,数据相符.678分析化学第33卷3.11.2合金模拟样中钴的测定移取～定量合金模拟样品溶液于10mL比色管中,在样品中加人3×10.mol/LF一来掩蔽Fe离子的干扰.以下操作同实验方法,按校准曲线进行定量,结果见表2.表2模拟样中钻的测定结果(n=6)Table2Determinationresultsofcobaltinsimulatedsamples(,l=6)合金模拟样溶液组成(thecompositionofsimulatedalloysample):1.Co1.0,Rh1.0,Pd1.0,Pt4.8,Cu2.0,Fe1.1,Ni7. 2;2.Co2.0,Rh1.0,Pd2.0,Pt5.0,Cu1.3.Fe1.1,Ni4.0(mg/L)References1LiuBin(刘彬),ZhangGuang(张光),ZhangXiaollng(张小玲).Chinese上Ana1.Chem.(分析化学),1995,23(8):9812ZhangMinghao(张明浩),ZhouChunshan(周春山),LiZhihong(李志红).ChineseAna1.Chem.(分析化学),20OO.28(8):997～10013ZhangShuyun(张淑云).AppliedChemistry(应用化学),1988.5(1):93-964WangY an(王艳),ZhouChunshan(周春山),JiangXinyu(蒋新宇).PhysicalTestingandChemicalAnalysis.尸B,t日:ChemicalAnalysis(理化检验-化学分册),2001.37(8):343～3455ZouY ongraing(邹永明),DingZhongtao(丁中涛),WangBingchang(王秉常).AnalyticalLaboratory(分析试验室),1995,14(2):26～286SafariA,ShamsE.Talanta,2000.51:1117～11237LiNanqiang(李南强),GaoXiaoxia(高小霞).Chinese上AnaloChem.(分析化学),1973,1(14):40～448NiY arning(倪亚明),LiLing(李玲),GaoXiaoxia(高小霞).ActaChimlcaSinica(化学),1988.46:651～6569GaoXiaoxia(高小霞).PolarographicCatalyticWave(极谱催化波).Bering(北京):SciencePress(科学出版社).1991:266～26710BandAJ,FaulknerLR.ElectrochemicalMethods.NewY ork:AcademicPress.1979:83～8511LiNanqiang(李南强),ZhaoY annan(赵燕南),GaoXiaoxia(高小霞).ActaChimicaSinica(化学),1984,42:1057～106212ZhangZuxun(张祖训),TuYifeng(屠一锋).Chem.ChineseUniversities(高等学校化学),1985.6(5):404～4JD813LavironE.Electroana1.Chem..1974,52:355～393 ComplexAdsorptiveWaveofco(n)-MethylThymolBlue-NO;andItsApplicationY angLizhu'_,ChenDahui,LiuChuanyin2,LiuWei.LuGuanghan'(SchoolP,WenzhouMedicalCollege,Wenzhou325027)(CollegeofChemistry,CentralChinaNormalUnivers~y,Wuhan430079)AbstractIn0.04mol/LNH,.H2O—NH4C1buffersolution,atpH8.5,asensitivesecondderivativepolar- ographicwaveofthecomplexofCo(1)一methylthymolblue(MTB)appearsat一1.23V(vs.SCE)inpresenceofNO;.Thepeakcu. rrentisproportionaltocobaltconcentrationintherangeof5.0×10～～2.0×10一mol/L,thedetectionlimitis1.0×l0一mol/L.Severalelectrochemicalmethodshavebeenusedtostudythe第5期杨丽珠等:钻(Ⅱ)?甲基百里香酚蓝络合剂.亚硝酸根络合吸附波的研究及应用679 propertiesofthepolarographicwaveandthemechanismofelectrode.Itisprovedthatthewave isacomplexadsorptiveone.epeakcurrentisproducedbythereductionofCo(n).Thesaturatedadsorption amountofthecomplexatHgelectrodeis5.18×10一moL/cmandtheadsorptionobeysFrumkinadsorptionisotherm. eadsorptioncoefficient(/3)is6.86×10andAGis33.30kJ/mo1.enumberofelectronstransferred(凡)is2andthetransfercoefficient()oftheirreversibleadsorptivesystemis0.58andtherateco nstantofapparentelectrodereaction(k.)is2.26S～.ismethodhasbeensuccessfullyappliedf0rthedet erminationoftracecobaltinvitaminB12andartificialsamples.KeywordsCobalt,methylthymolbluecomplexone,nitrite,complexadsorptivewave (Received1March2004;accepted9June2004)<<<<<<<<<<<<<<<<<<<<< <<<<<<<<<<<<<<<<<<<<< <<<<<<<<<<<<<<<<<<<<< <<<<<<<<<<<<<<<<<<<<< <<<<《分析化学》入选2004年度"CA千种表"根据美国化学会出版的{ChemicalAbstractsServiceSourceIndexQuartedyNo.4)统计结果.2004年度我国(包括台湾地区)有94种科技期刊进入"CA千种表".《分析化学)位居其中,现将入选期刊(前4o名)列出如下:CA千种表CA千种表序号刊名序号刊名名次名次1高等学校化学13721无机化学3972世界胃肠病学杂志(英文版)19122中国塑料3983第四军医大学2Q523中国药理(英文版)4204分析化学21724中国有色金属4465化学23625物理化学4596第三军医大学25526中国病理生理杂志4677光谱学与光谱分析26627功能材料4748中国化学快报(英文版)28628中国生物工程杂志4809世界华人消化杂志28729石油化工48610钢铁29330化学通报494l1中国给水排水29431精细化工5O212化工29632硅酸盐5O913物理3O233分析试验室53514高分子材料科学与工程34134细胞与分子免疫学杂志54015科学通报(英文版)34535中国有色金属学会会刊(英文版)54416环境污染治理技术与设备34836第一军医大学54517中国化学(英文版)35537稀有金属材料与工程55618有机化学38138中国化学会会志(英文版,台北)56819光谱实验室38239中华医学杂志(英文版)57620应用化学39140色谱579厦门大学化学化工学院资料室黄秀菁提供。

全反式维甲酸影响JAK2/STAT3信号通路抑制肺纤维化EMT发布时间：2021-09-02T10:31:02.507Z 来源：《中国结合医学杂志》2021年2期作者：贾闪闪侯婷婷娄晓月苗薇薇刘巨源[导读] 目的探讨全反式维甲酸贾闪闪侯婷婷娄晓月苗薇薇刘巨源新乡医学院三全学院，河南省新乡453003摘要：目的探讨全反式维甲酸（ATRA）对肺泡上皮细胞（AEC）向间质细胞转化（EMT）的影响及分子机制。

方法分离大鼠的AEC II进行原代细胞培养，取5~7代细胞分为正常对照组、5μg/L TGF-β1诱导组、10μmol/L ATRA组、5μg/L TGF-β1诱导+0.1μmol/L ATRA组、5μg/L TGF-β1诱导+1μmol/L ATRA组、5μg/L TGF-β1诱导+10 μmol/L ATRA组，37℃ 5% CO2条件下培养24h，RT-PCR方法检测细胞中Collagen III、α-SMA、JAK2、STAT3和SOCS3 mRNA水平，37℃ 5% CO2条件下培养72h，Western Blot检测细胞中α-SMA、JAK2、STAT3和SOCS3蛋白表达水平。

结果和空白组相比，TGF诱导组的α-SMA mRNA和蛋白表达水平均升高，Collagen III、JAK2、STAT3 mRNA水平显著升高，SOCS mRNA水平显著降低，JAK2、STAT3、pSTAT3蛋白水平均显著升高，差异有统计学意义（P<0.05）；和TGF诱导组相比，TGF-β1诱导+10 μmol/L ATRA组和TGF-β1诱导+1 μmol/L ATRA组的α-SMA mRNA和蛋白表达水平均降低，Collagen III、JAK、STAT3 mRNA水平显著降低，SOCS mRNA水平显著升高，JAK2、STAT3、pSTAT3蛋白水平显著降低，差异有统计学意义（P<0.05）。

LC-MS检测西他沙星原料中基因毒性杂质的含量石莹1宋雪洁3李浩冬2路显锋2*1药物研究院分析所，扬子江药业集团，泰州2253212药物制剂新技术国家重点实验室，扬子江药业集团，泰州2253213质量管理部，扬子江药业集团，泰州225321摘要建立了LC-MS 法测定西他沙星中基因毒性杂质对甲苯磺酸甲酯和对甲苯磺酸乙酯含量的方法。

方法:采用Agilent Poroshell 120 EC-C18色谱柱；流动相为纯水（0.1%甲酸）:甲醇（V/V）=60:40；稀释剂为乙腈（0.1%甲酸）:纯水（V/V）=50:10；柱温为40℃；进样体积为5µl；流速为0.4ml/min；采用正离子模式进行扫描。

对甲苯磺酸甲酯测定浓度在0.76ng/ml~15.27ng/ml范围内，线性关系良好；对甲苯磺酸乙酯测定浓度在0.75ng/ml~15.01ng/ml范围内，线性关系良好。

对甲苯磺酸甲酯的定量限为0.0038ng；对甲苯磺酸乙酯的定量限为0.0038ng。

杂质回收率在限度浓度80%、100%和160%三个浓度水平均在90~110%之间，该方法准确度良好。

该方法适用于西他沙星原料中对甲苯磺酸甲酯和对甲苯磺酸乙酯的检测。

西他沙星(sitafloxacin)是日本第一制药有限公司继左氧氟沙星后开发出的一种强力广谱新氟喹诺酮类抗菌剂，该药对革兰氏阳性球菌，革兰氏阴性菌以及厌氧菌的抗菌活性是左氧氟沙星的4～32倍，同时对肺炎球菌DNA 促旋酶和拓扑同功酶有双重抑制作用。

临床表现有极广的抗菌谱，特别是对呼吸道的病菌有极强的抗菌活性。

因西他沙星的一个起始物料为对甲苯磺酸盐，在后续反应中对甲苯磺酸若有残留，可能会与溶剂甲醇、乙醇反应生成具有基因毒性的杂质—对甲苯磺酸甲酯和对甲苯磺酸乙酯，故采用LC-MS法对产品中的对甲苯磺酸甲酯/乙酯进行控制。

1、实验部分1.1仪器与试药Agilent 1200液相色谱仪(美国安捷伦公司)；Agilent 6460三重串联四极杆质谱仪(美国安捷伦公司)；XP205型电子天平(瑞士梅特勒托利多公司)。

泽泻醇在小胶质细胞中对基质金属蛋白酶3与一氧化氮的抑制作用刘瑜【摘要】This paper investigates the inhibitory effect and mechanism of alismol on neuroinflamination in the activated BV2 microglial cells which are stimulated by lipopolysaccharides(LPS). NO was measured by using Griess reagent. RT-PCR and Western blot are used to analyse ERK, JNK, Akt, and MMP3 . Alismol can significantly inhibit LPS-induced NO production and the MMP3 expression. The mechanism is involved to its inhibition of PI3K/Akt pathway.%利用脂多糖(LPS)刺激小鼠小胶质细胞BV2,研究泽泻醇对炎症相关分子的抑制及机制.Griess法测定一氧化氮(NO)浓度,RT-PCR和Western blot法检测细胞外调节蛋白激酶(ERK)、p38、c-Jun氨基末端激酶(JNK)、蛋白激酶B(Akt)、基质金属蛋白酶3(MMP3)的变化.研究结果表明,泽泻醇不仅对LPS刺激小胶质细胞产生的NO有明显抑制作用,还能在mRNA与蛋白质水平抑制MMP3的表达,这种抑制与其对PI3K/Akt通路的干预相关.阐述了泽泻醇对小胶质细胞的抑制与PI3K/Akt通路的相关机制.【期刊名称】《实验技术与管理》【年(卷),期】2012(029)010【总页数】4页(P47-50)【关键词】小胶质细胞;泽泻醇;一氧化氮;基质金属蛋白酶3【作者】刘瑜【作者单位】南开大学医学院,天津 300071【正文语种】中文【中图分类】R914Abstract：This paper investigates the inhibitory effect and mechanism of alismol on neuroinflammation in the activated BV2microglial cells whichare stimulated by lipopolysaccharides（LPS）.NO was measured by using Griess reagent.RT-PCR and Western blot are used to analyse ERK，JNK，Akt，and MMP3 .Alismol can significantly inhibit LPS-induced NO production and the MMP3expression.The mechanism is involved to its inhibition of PI3K/Akt pathway.Key words：microglia；alismol；NO；MMP3小胶质细胞是中枢神经系统内的免疫细胞，长期激活而形成中枢神经系统的慢性炎症，其释放的大量氧自由基、炎症介质细胞因子以及基质金属蛋白酶（matrix metalloproteinases，MMP）是神经元损伤的重要原因之一，也是阿尔茨海默病（Alzheimer’s disease，AD）与帕金森病（Parkinson’s disease，PD）等许多中枢神经退行性疾病发生与发展的重要因素之一［1-2］。

Inhibitors, Agonists, Screening Libraries Data SheetBIOLOGICAL ACTIVITY:Alda–1 is a potent ALDH2 agonist, which significantly improves ALDH2 activity.In Vivo: Alda–1 treatment results in a significant decrease of 4–HNE–protein content in the plasma of apoE -/- mice. Alda–1administration leads to a slight increase in gene expression related to neurogenesis (Nog ), mitochondrial biogenesis (CYTB , ND1),and apoptosis (Bax , Gsk3b ) in the Hp of apoE -/- mice. Alda–1 administration leads to 2 and 10 differentially expressed proteins in theFCx and Hp of apoE -/- mice, respectively [1]. Alda–1 (1.5 mg/kg, b.w., i.p.) administration significantly increases the climbing time,tends to reduce the immobility time and increases the swimming time of the prenatally stressed rats in the forced swim test.Moreover, treatment of prenatally stressed rats with Alda–1 significantly increases number of entries into the open arms of the maze and the time spent therein, as assessed by elevated plus–maze test [2]. Alda–1 (8.5 mg/kg, i.p.) with glucose significantly lowers 4–HNE and FJB–positive cells in the cerebral cortex of Alda–1–treated rats than in DMSO–treated rats 24 h after glucose administration [3]. Alda–1 (10 mg/kg per day) treatment prevents aldehydic overload, mitochondrial dysfunction and improves ventricular function in post–MI cardiomyopathy rats [4].PROTOCOL (Extracted from published papers and Only for reference)Cell Assay:[2]Spleen cells (4×106 cells/mL) are stimulated by optimal concentrations of concanavalin A (Con A; 2.5μg/mL and 0.6 μg/mL) and lipopolysaccharide (LPS, 5 μg/mL) and are incubated in 96–well plates at final volume of 0.2mL for 72 h. Cell proliferation is determined by adding 0.5 μCi of [3H]–thymidine per well at 16 h before the end of the incubation.The cultures are harvested with an automatic cell harvester, and [3H] thymidine incorporation is assessed using a liquid scintillationcounter.Animal Administration: Alda–1 is dissolved in 1 mL/kg b.w. DMSO/water 50/50.[2]After behavioral verification at three months of age,the animals are divided into the following four groups: control, control + Alda–1, prenatally stressed and prenatally stressed + Alda–1(6 animals per group). Alda–1 injections are given intraperitoneally (i.p.) once daily at a dose of 1.5 mg/kg b.w. (dissolved in 1 mL/kg b.w. DMSO/water 50/50) for 14 days. At the same time, the control and prenatally stressed rats receive 1 mL/kg b.w. DMSO/water 50/50. The injections of Alda–1 and vehicle are given between 10 a.m and 11 a.m. In the last five days of Alda–1 treatment the behavioral parameters in the elevated plus maze test and then in the forced swim test are measured.References:[1]. Stachowicz A, et al. Proteomic Analysis of Mitochondria–Enriched Fraction Isolated from the Frontal Cortex and Hippocampus of Apolipoprotein E Knockout Mice Treated with Alda–1, an Activator of Mitochondrial Aldehyde Dehydrogenase (ALDH2). Int J Mol Sci.[2]. Stachowicz A, et al. The impact of mitochondrial aldehyde dehydrogenase (ALDH2) activation by Alda–1 on the behavioral and biochemical disturbances in animal model of depression. Brain Behav Immun. 2016 Jan;51:144–53.Product Name:Alda–1Cat. No.:HY-18936CAS No.:349438-38-6Molecular Formula:C 15H 11Cl 2NO 3Molecular Weight:324.16Target:Aldehyde Dehydrogenase (ALDH)Pathway:Metabolic Enzyme/Protease Solubility:DMSO: ≥ 51 mg/mL[3]. Ikeda T, et al. Effects of Alda–1, an Aldehyde Dehydrogenase–2 Agonist, on Hypoglycemic Neuronal Death. PLoS One. 2015 Jun 17;10(6):e0128844.[4]. Gomes KM, et al. Aldehydic load and aldehyde dehydrogenase 2 profile during the progression of post–myocardial infarction cardiomyopathy: benefits of Alda–1. Int J Cardiol. 2015 Jan 20;179:129–138.Caution: Product has not been fully validated for medical applications. For research use only.Tel: 609-228-6898 Fax: 609-228-5909 E-mail: tech@Address: 1 Deer Park Dr, Suite Q, Monmouth Junction, NJ 08852, USA。

=====================================================================Acq. Operator : Li Shan(LCMS-02) Seq. Line : 62Acq. Instrument : HY-LCMS-02 Location : P1-C-05Injection Date : 7/1/2015 5:33:46 PM Inj : 1Inj Volume : 3.000 µlAcq. Method : D:\AGLIENT 1260\DATA\20150701-1\20150701-1 2015-07-01 12-16-29\100-1000MS+ 3MIN-1.5_(0.02%FA).MLast changed : 7/1/2015 12:16:29 PM by Li Shan(LCMS-02)Analysis Method : D:\AGLIENT 1260\METHOD\100-1500MS+7MIN_1.5(0.02%FA).M Last changed : 7/2/2015 1:12:31 PM by Li Shan(LCMS-02) (modified after loading)Method Info : Postive,MS:100-1000,Column ID:A-RP-85,50℃Catalog No : HY-12832 Batch#16920 A-RP-132Additional Info : Peak(s) manually integratedmin0.511.52 2.53mAU 02505007501000125015001750 DAD1 B, Sig=214,4 Ref=off (D:\AGLIENT...TA\20150701-1\20150701-1 2015-07-01 12-16-29\BIZ2015-701-DJL16.D)1.7902.0042.130===================================================================== Area Percent Report =====================================================================Sorted By : Signal Multiplier : 1.0000Dilution : 1.0000Do not use Multiplier & Dilution Factor with ISTDsSignal 1: DAD1 B, Sig=214,4 Ref=offPeak RetTime Type Width Area Height Area # [min] [min] [mAU*s] [mAU] %----|-------|----|-------|----------|----------|--------| 1 1.790 MM 0.0989 2.54602 4.29243e-1 0.0254 2 2.004 MM 0.0610 4.59333 1.25517 0.0458 3 2.130 MM 0.0836 1.00147e4 1997.11499 99.9288Totals : 1.00218e4 1998.79940===================================================================== *** End of Report ***=====================================================================Acq. Operator : Li Shan(LCMS-02) Seq. Line : 62Acq. Instrument : HY-LCMS-02 Location : P1-C-05Injection Date : 7/1/2015 5:33:46 PM Inj : 1Inj Volume : 3.000 µlAcq. Method : D:\AGLIENT 1260\DATA\20150701-1\20150701-1 2015-07-01 12-16-29\100-1000MS+ 3MIN-1.5_(0.02%FA).MLast changed : 7/1/2015 12:16:29 PM by Li Shan(LCMS-02)Analysis Method : D:\AGLIENT 1260\METHOD\100-1500MS+7MIN_1.5(0.02%FA).M Last changed : 7/2/2015 1:13:35 PM by Li Shan(LCMS-02) (modified after loading)Method Info : Postive,MS:100-1000,Column ID:A-RP-85,50℃Catalog No : HY-12832 Batch#16920 A-RP-132Additional Info : Peak(s) manually integratedmin0.511.522.5350000100000150000200000250000300000 MSD1 TIC, MS File (D:\AGLIENT 1260\DATA\20150701-1\20150701-1 2015-07-01 12-16-29\BIZ2015-701-DJL16.D) ES-API, Pos2.140MS Signal: MSD1 TIC, MS File, ES-API, Pos, Scan, Frag: 50 Spectra averaged over upper half of peaks. Noise Cutoff: 1000 counts.Reportable Ion Abundance: > 10%.Retention Mol. Weight Time (MS) MS Area or Ion2.140 2099302 349.00 I 347.95 I 347.00 Im/z10015020025030035040045050020406080100*MSD1 SPC, time=2.108:2.181 of D:\AGLIENT 1260\DATA\20150701-1\20150701-1 2015-07-01 12-16-29\BIZ2015-701-DJL16.D ES Max: 159731446.2348.0349.0 347.0*** End of Report ***。