筛选靶向结合炎症相关蛋白5-LOX(5-lipoxygenase,5-脂氧合酶)的中药黄酮类天然产物,分析与5-LOX结合的黄酮类成分及其来源中药的共性规律。本研究借助Discovery Studio 2017 R2分子对接和药效团构建模块,结合SPR分子筛选实验,以及关联...筛选靶向结合炎症相关蛋白5-LOX(5-lipoxygenase,5-脂氧合酶)的中药黄酮类天然产物,分析与5-LOX结合的黄酮类成分及其来源中药的共性规律。本研究借助Discovery Studio 2017 R2分子对接和药效团构建模块,结合SPR分子筛选实验,以及关联网络构建的方法进行研究。研究结果显示,来源于17种中药的18个黄酮类小分子中有11个能够与5-LOX结合,并从分子对接以及药效团构建研究中发现其作用的3种方式和共性特征:(1)部分中药黄酮成分(如木犀草素等)通过结构中的B环与5-LOX在活性位点ASP243形成静电中心相结合;(2)部分中药黄酮成分(如芹菜素等)是通过结构中的A环与活性位点VAL520形成疏水键、与活性位点ASP243形成氢键与5-LOX结合;(3)杨梅苷等黄酮类成分由于极性较强,在没有形成疏水键的情况下,也是通过形成静电中心与5-LOX在活性位点ASP243产生相互作用。此外还发现靶向5-LOX的活性中药黄酮类化合物,大多来源于具有利湿、退黄等功效,性味甘苦寒的景天科中药中。本研究发现了部分靶向5-LOX的中药黄酮类成分及其作用方式和共性规律,为开发靶向5-LOX抗肿瘤新药提供思路和方法。展开更多
利用网络药理学和分子对接方法探究玫瑰花抗抑郁的物质基础及作用机制。通过中国知网收集抗抑郁单体和玫瑰花的化学成分,借助PubChem、SwissTargetPrediction数据库预测玫瑰花的抗抑郁活性成分及作用靶点,通过DAVID数据库进行通路富集分...利用网络药理学和分子对接方法探究玫瑰花抗抑郁的物质基础及作用机制。通过中国知网收集抗抑郁单体和玫瑰花的化学成分,借助PubChem、SwissTargetPrediction数据库预测玫瑰花的抗抑郁活性成分及作用靶点,通过DAVID数据库进行通路富集分析,运用Discovery Studio 2017软件对关键成分和靶点进行分子对接。收集抗抑郁单体117个,玫瑰花成分127个,筛选出玫瑰花与抗抑郁相关的核心成分17个,分子对接显示玫瑰花所含芹菜素、槲皮素、山柰酚、异丁香酚、异槲皮苷、矢车菊素-3-O-葡萄糖苷6个活性成分,可较好地与AKT1、PTGS2、ESR13个抑郁相关靶点结合。结果表明玫瑰花抗抑郁的物质基础为芹菜素、槲皮素、山柰酚等黄酮类化合物,主要通过影响HIF-1信号通路、神经活性配体-受体相互作用信号通路、催乳激素信号通路、多巴胺能神经突触信号通路发挥抗抑郁作用。该研究为抗抑郁新药研发提供了理论依据及新的研究思路。展开更多
Objective:To explore the potential mechanism of lysionotin in treating glioma.Methods:First,target prediction based on Bernoulli Naïve Bayes profiling and pathway enrichment was used to predict the biological act...Objective:To explore the potential mechanism of lysionotin in treating glioma.Methods:First,target prediction based on Bernoulli Naïve Bayes profiling and pathway enrichment was used to predict the biological activity of lysionotin.The binding between 5-lipoxygenase(5-LO)and lysionotin was detected by surface plasmon resonance(SPR)and molecular docking,and the inhibitory effects of lysionotin on 5-LO and proliferation of glioma were determined using enzyme inhibition assay in vitro and cell viability analysis,respectively.Furthermore,the pharmaceutical effect of lysionotin was explored by cell survival rate analysis and liquid chromatography with tandem mass spectrometry(LC-MS/MS).The protein expression,intracellular calcium ion concentration and cytoskeleton detection were revealed by Western blot,flow cytometry and fluorescence labeling,respectively.Results:Target prediction and pathway enrichment revealed that lysionotin inhibited 5-LO,a key enzyme involved in the arachidonic acid metabolism pathway,to inhibit the proliferation of glioma.Molecular docking results demonstrated that 5-LO can be binding to lysionotin through hydrogen bonds,forming bonds with His600,Gln557,Asn554,and His372.SPR analysis further confirmed the interaction between 5-LO and lysionotin.Furthermore,enzyme inhibition assay in vitro and cell survival rate analysis revealed that 50%inhibition concentration of lysionotin and the median effective concentration of lysionotin were 90 and 16.58µmol/L,respectively,and the results of LC-MS/MS showed that lysionotin inhibited the production of 5S-hydroperoxy-eicosatetraenoic acid(P<0.05),and moreover,the LC-MS/MS results indicated that lysionotin can enter glioma cells well(P<0.01)and inhibit their proliferation.Western blot analysis demonstrated that lysionotin can inhibit the expression of 5-LO(P<0.05)and downstream leukotriene B4 receptor(P<0.01).In addition,the results showed that lysionotin affected intracellular calcium ion concentration by inhibiting 5-LO to affect the cytoskeleton,as determined by flow cytometry and fluorescence labeling.Conclusion:Lysionotin binds to 5-LO could suppress glioma by inhibiting arachiodonic acid metabolism pathway.展开更多
Background In vitro chondrocyte expansion is a major challenge in cell-based therapy for human articular cartilage repair. Classical culture conditions usually use animal serum as a medium supplement, which raises a n...Background In vitro chondrocyte expansion is a major challenge in cell-based therapy for human articular cartilage repair. Classical culture conditions usually use animal serum as a medium supplement, which raises a number of undesirable questions. In the present study, two kinds of defined, serum-free media were developed to expand chondrocytes in monolayer culture for the purpose of cartilage tissue engineering. Methods Bovine chondrocytes were expanded in serum-free media supplemented with fibroblast growth factor-2 and platelet-derived growth factor or fibroblast growth factor-2 and insulin-like growth factor. Expansion culture in a conventional 10% fetal bovine serum （FBS） medium served as control. Fibronectin coating was used to help cell adhesion in serum-free medium. Next, in vitro three-dimensional pellet culture was used to evaluate the chondrocyte capacity. Cell pellets were expanded in different media to re-express the differentiated phenotype （re-differentiation） and to form cartilaginous tissue. The pellets were assessed by glycosaminoglycans contents, collagen II, collagen I and collagen X immunohistological staining. Results Chondrocytes cultured in serum-free media showed no proliferation difference than cells grown with 10% FBS medium. In addition, chondrocytes expanded in both serum-free media expressed more differentiated phenotypes at the end of monolayer culture, as indicated by higher gene expression ratios of collagen type II to collagen type I. Pellets derived from chondrocytes cultured in both serum-free media displayed comparable chondrogenic capacities to pellets from cells expanded in 10% FBS medium. Conclusion These findings provide alternative culture approaches for chondrocytes in vitro expansion, which may benefit the clinical use of autologous chondrocytes implantation.展开更多
Two novel substituted phenyl isoxazole benzoxazine formamide derivatives were designed and synthesized with substituted o-aminophenol,1,2-dibromoethane and different phenyl substituted isoxazole formyl chloride as the...Two novel substituted phenyl isoxazole benzoxazine formamide derivatives were designed and synthesized with substituted o-aminophenol,1,2-dibromoethane and different phenyl substituted isoxazole formyl chloride as the raw materials via microwave assistant synthesis.The target compounds were characterized by IR,^(1)H NMR,^(13)C NMR and HRMS.Both single-crystal structures were further determined by X-ray diffraction.3-(2’-Chloro-6’-fluoro-phenyl)-4-(2’,3’-dihydro-1’,4’-benzoxazine)-5-methyl-isoxazole formamide (4a) crystallizes in orthorhombic system,P2_(1) space group with a=8.9414(18),b=10.834(2),c=17.706(4)A,V=1715.1(6)A^(3),D_(c)=1.444 Mg/m^(3),Z=4,F(000)=768,μ(Mo Kα)=0.255 mm^(-1),R=0.0406 and wR=0.1171.3-Phenyl-4-(6-methyl-2’,3’-dihydro-1’,4’-benzoxazine)-5-methyl-isoxazole formamide (4b) is of triclinic system,space group PI with a=7.7659(16),b=8.3626(17),c=13.484(3)A,α=76.04(3)°,β=100.63(3)°,γ=82.01(3)°,V=841.6(3)A^(3),D_(c)=1.319 Mg/m^(3),Z=2,F(000)=352,μ(Mo Kα)=0.090 mm^(-1),R=0.0672 and wR=0.2671.Both crystals are packed through C–H···O hydrogen bonding interaction.There is C–H···F hydrogen bond between 4a molecules,and C–H···N between 4b molecules.Bioassay results showed that compounds 4a and 4b exhibited detoxification on maize and restored maize growth index.展开更多
文摘筛选靶向结合炎症相关蛋白5-LOX(5-lipoxygenase,5-脂氧合酶)的中药黄酮类天然产物,分析与5-LOX结合的黄酮类成分及其来源中药的共性规律。本研究借助Discovery Studio 2017 R2分子对接和药效团构建模块,结合SPR分子筛选实验,以及关联网络构建的方法进行研究。研究结果显示,来源于17种中药的18个黄酮类小分子中有11个能够与5-LOX结合,并从分子对接以及药效团构建研究中发现其作用的3种方式和共性特征:(1)部分中药黄酮成分(如木犀草素等)通过结构中的B环与5-LOX在活性位点ASP243形成静电中心相结合;(2)部分中药黄酮成分(如芹菜素等)是通过结构中的A环与活性位点VAL520形成疏水键、与活性位点ASP243形成氢键与5-LOX结合;(3)杨梅苷等黄酮类成分由于极性较强,在没有形成疏水键的情况下,也是通过形成静电中心与5-LOX在活性位点ASP243产生相互作用。此外还发现靶向5-LOX的活性中药黄酮类化合物,大多来源于具有利湿、退黄等功效,性味甘苦寒的景天科中药中。本研究发现了部分靶向5-LOX的中药黄酮类成分及其作用方式和共性规律,为开发靶向5-LOX抗肿瘤新药提供思路和方法。
文摘利用网络药理学和分子对接方法探究玫瑰花抗抑郁的物质基础及作用机制。通过中国知网收集抗抑郁单体和玫瑰花的化学成分,借助PubChem、SwissTargetPrediction数据库预测玫瑰花的抗抑郁活性成分及作用靶点,通过DAVID数据库进行通路富集分析,运用Discovery Studio 2017软件对关键成分和靶点进行分子对接。收集抗抑郁单体117个,玫瑰花成分127个,筛选出玫瑰花与抗抑郁相关的核心成分17个,分子对接显示玫瑰花所含芹菜素、槲皮素、山柰酚、异丁香酚、异槲皮苷、矢车菊素-3-O-葡萄糖苷6个活性成分,可较好地与AKT1、PTGS2、ESR13个抑郁相关靶点结合。结果表明玫瑰花抗抑郁的物质基础为芹菜素、槲皮素、山柰酚等黄酮类化合物,主要通过影响HIF-1信号通路、神经活性配体-受体相互作用信号通路、催乳激素信号通路、多巴胺能神经突触信号通路发挥抗抑郁作用。该研究为抗抑郁新药研发提供了理论依据及新的研究思路。
基金Supported by the Natural Science Foundation of China(No.81473369)the National Key Research and Development Program of China(No.2017YFC1702703)Jinan City Science and Technology SMES Innovation Ability Improvement Project,“Traditional Chinese Medicine Digital Humanities Youth Innovation Team”of College and University in Shandong Province(No.2023RW093)。
文摘Objective:To explore the potential mechanism of lysionotin in treating glioma.Methods:First,target prediction based on Bernoulli Naïve Bayes profiling and pathway enrichment was used to predict the biological activity of lysionotin.The binding between 5-lipoxygenase(5-LO)and lysionotin was detected by surface plasmon resonance(SPR)and molecular docking,and the inhibitory effects of lysionotin on 5-LO and proliferation of glioma were determined using enzyme inhibition assay in vitro and cell viability analysis,respectively.Furthermore,the pharmaceutical effect of lysionotin was explored by cell survival rate analysis and liquid chromatography with tandem mass spectrometry(LC-MS/MS).The protein expression,intracellular calcium ion concentration and cytoskeleton detection were revealed by Western blot,flow cytometry and fluorescence labeling,respectively.Results:Target prediction and pathway enrichment revealed that lysionotin inhibited 5-LO,a key enzyme involved in the arachidonic acid metabolism pathway,to inhibit the proliferation of glioma.Molecular docking results demonstrated that 5-LO can be binding to lysionotin through hydrogen bonds,forming bonds with His600,Gln557,Asn554,and His372.SPR analysis further confirmed the interaction between 5-LO and lysionotin.Furthermore,enzyme inhibition assay in vitro and cell survival rate analysis revealed that 50%inhibition concentration of lysionotin and the median effective concentration of lysionotin were 90 and 16.58µmol/L,respectively,and the results of LC-MS/MS showed that lysionotin inhibited the production of 5S-hydroperoxy-eicosatetraenoic acid(P<0.05),and moreover,the LC-MS/MS results indicated that lysionotin can enter glioma cells well(P<0.01)and inhibit their proliferation.Western blot analysis demonstrated that lysionotin can inhibit the expression of 5-LO(P<0.05)and downstream leukotriene B4 receptor(P<0.01).In addition,the results showed that lysionotin affected intracellular calcium ion concentration by inhibiting 5-LO to affect the cytoskeleton,as determined by flow cytometry and fluorescence labeling.Conclusion:Lysionotin binds to 5-LO could suppress glioma by inhibiting arachiodonic acid metabolism pathway.
文摘Background In vitro chondrocyte expansion is a major challenge in cell-based therapy for human articular cartilage repair. Classical culture conditions usually use animal serum as a medium supplement, which raises a number of undesirable questions. In the present study, two kinds of defined, serum-free media were developed to expand chondrocytes in monolayer culture for the purpose of cartilage tissue engineering. Methods Bovine chondrocytes were expanded in serum-free media supplemented with fibroblast growth factor-2 and platelet-derived growth factor or fibroblast growth factor-2 and insulin-like growth factor. Expansion culture in a conventional 10% fetal bovine serum （FBS） medium served as control. Fibronectin coating was used to help cell adhesion in serum-free medium. Next, in vitro three-dimensional pellet culture was used to evaluate the chondrocyte capacity. Cell pellets were expanded in different media to re-express the differentiated phenotype （re-differentiation） and to form cartilaginous tissue. The pellets were assessed by glycosaminoglycans contents, collagen II, collagen I and collagen X immunohistological staining. Results Chondrocytes cultured in serum-free media showed no proliferation difference than cells grown with 10% FBS medium. In addition, chondrocytes expanded in both serum-free media expressed more differentiated phenotypes at the end of monolayer culture, as indicated by higher gene expression ratios of collagen type II to collagen type I. Pellets derived from chondrocytes cultured in both serum-free media displayed comparable chondrogenic capacities to pellets from cells expanded in 10% FBS medium. Conclusion These findings provide alternative culture approaches for chondrocytes in vitro expansion, which may benefit the clinical use of autologous chondrocytes implantation.
基金the National Natural Science Foundation of China(No.31801784 and 31572042)Undergraduate SIPT program of Northeast Agricultural University(No.202010224112)。
文摘Two novel substituted phenyl isoxazole benzoxazine formamide derivatives were designed and synthesized with substituted o-aminophenol,1,2-dibromoethane and different phenyl substituted isoxazole formyl chloride as the raw materials via microwave assistant synthesis.The target compounds were characterized by IR,^(1)H NMR,^(13)C NMR and HRMS.Both single-crystal structures were further determined by X-ray diffraction.3-(2’-Chloro-6’-fluoro-phenyl)-4-(2’,3’-dihydro-1’,4’-benzoxazine)-5-methyl-isoxazole formamide (4a) crystallizes in orthorhombic system,P2_(1) space group with a=8.9414(18),b=10.834(2),c=17.706(4)A,V=1715.1(6)A^(3),D_(c)=1.444 Mg/m^(3),Z=4,F(000)=768,μ(Mo Kα)=0.255 mm^(-1),R=0.0406 and wR=0.1171.3-Phenyl-4-(6-methyl-2’,3’-dihydro-1’,4’-benzoxazine)-5-methyl-isoxazole formamide (4b) is of triclinic system,space group PI with a=7.7659(16),b=8.3626(17),c=13.484(3)A,α=76.04(3)°,β=100.63(3)°,γ=82.01(3)°,V=841.6(3)A^(3),D_(c)=1.319 Mg/m^(3),Z=2,F(000)=352,μ(Mo Kα)=0.090 mm^(-1),R=0.0672 and wR=0.2671.Both crystals are packed through C–H···O hydrogen bonding interaction.There is C–H···F hydrogen bond between 4a molecules,and C–H···N between 4b molecules.Bioassay results showed that compounds 4a and 4b exhibited detoxification on maize and restored maize growth index.
