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A system for screening agonists targeting β_2-adrenoceptor from Chinese medicinal herbs

Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2009年第4期10卷 243-250页

作者： Hui WANG Shi-you LI Chuan-ke ZHAO Xin ZENG Graduate School of the Chinese Academy of Sciences Beijing 100039 China Beijing Institute of Genomics Chinese Academy of Sciences Beijing 100029 China

In order to develop a model for screening the agonists of human β2-adrenoceptor from Chinese medicinal herbs extracts, we used a cell-based functional assay based on a common G protein-coupled receptor （GPCR） regulation mechanism and destabilized enhanced green fluorescent protein （d2EGFP） reporter gene technique. The positive cell clone was confirmed by real-time polymerase chain reaction （PCR） and imaging analysis. To assess the value of this model, we screened over 2000 high performance liquid chromatography （HPLC）-fractionated samples from the ethanol extracts of Chinese medicinal herbs. Six fractions （isolated from Panax japonicus, Veratrum nigrum, Phellodendron amurense, Fructus Aurantii lmmaturus, Chaenomeles speciosa, and Dictamnus dasycarpus） showed significant effects on active reporter gene expression, three of which （isolated from Phellodendron amurense, Fructus Aurantii lmmaturus, and Chaenomeles speciosa） were selected for further concentration response analysis and the half maximal effective concentration （EC1/2 max） values were 4.2, 2.7, and 4.8 μg/ml, respectively. Therefore, this reporter gene assay was suitable for screening β2-adrenoceptor agonists. The results suggest that the six herbal extracts are the possible agonists of β2-adrenoceptor.

关键词： β2-adrenoceptor Enhanced green fluorescent protein (EGFP) Chinese medicinal herbs Screening

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Evidence for a Non-β2-adrenoceptor Binding Site in Human Lung Tissue for a Subset of β2-adrenoceptor Agonists

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Pharmacology & Pharmacy 2014年第1期5卷 30-36页

作者： Robert J. Slack Respiratory TAU Biology GlaxoSmithKline Stevenage UK.

The aim of this study was to compare the binding profile of a range of β2-adrenoceptor (β2-AR) agonists and antagonists in human lung tissue. Radioligand saturation and competition binding experiments were performed by filtration with a β2-AR antagonist ([3H]propranolol) or agonist ([3H]vilanterol) radioligand and membrane fragments generated from lung parenchyma in the presence of 100 μM guanosine 5’-[β,γ-imido]triphosphate (Gpp(NH)p). In membranes prepared from human lung parenchyma, carmoterol, formoterol, ICI118551, propranolol and salbutamol resulted in inhibition of [3H]vilanterol binding to levels that were significantly different from indacaterol, salmeterol and vilanterol (ANOVA, Bonferroni post-test, P 0.05). Indacaterol, salmeterol and vilanterol resulted in full inhibition of [3H]propranolol binding to levels not significantly different from ICI118551 (ANOVA, Bonferroni post-test, P > 0.05). Indacaterol, salmeterol and vilanterol bind to an additional site in human lung parenchyma membranes that is distinct from the β2-AR.

关键词： β2-adrenoceptor Radioligand Binding Human Lung Tissue Binding Site

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Primary study on correlation betweenβ_2-adrenoceptor haplotypes and asthma in children of Han nationality in Chongqing

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Journal of Medical Colleges of PLA(China) 2001年第4期16卷 299-302页

作者：廖伟李为明赵聪敏奚敏艾友萍温恩懿 Department of Pediatrics Xinqiao Hospital Third Military Medical University Changing 400037 China

Objective:To investigatethecorrelationbetweenβ 2 -adrenergicreceptors(β 2 -AR)haplotypesandasthmaof Hannationalitychildrenin ***:PCRandrestrictionfragmentanalysiswereusedto study16,27lociof theβ 2 -ARpolymorphismin76unrelatedasthmaticchildrenandin100healthychildrenandadultsof Hannationali-ty as control.A statisticalanalysisof thecorrelationbetweenglycine(Gly)16allele,Gly16/glutamine(Gln)27haplotype andasthmaticclinicalstatuswas ***:Therewas no significantincreaseof thefrequencyof Gly16and Gln27alleleintheasthmaticgroupas comparedwiththecontrolgroup(P>0.05).Therewasa significantincreaseof the frequencyof Gly16alleleandGly16/Gln27haplotypein severeasthmaticcasesthanin themildandmoderateasthmatic ones(P2 -***16alleleandGly16/Gln27haplotypearepossiblycorrelatedwiththeseverityof theclinicalmanifestationsin thechildrenof HannationalityinChongqing.

关键词： asthma children β2-adrenoceptor haplotype

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Effect of overexpression of β2-adrenoceptor on myocellular contraction in rat cardiomyocytes injured by isoprenoline

Effect of overexpression of β2-adrenoceptor on myocellular ...

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中国生理学会第23届全国会员代表大会暨生理学学术大会

作者： Chuan-Ying XU,Xin-Wei JIANG,Ying WANG,Guang-Ming WANG,Li-Ping GAO,Ai-Ying LIU,Hong SUN~* Physiology Department of Xuzhou Medical College,Xuzhou 221002,China

Aim:To investigate whether overexpression ofβ-adrenoceptor(β-AR) could improve myocellular contraction in ventricular cardiomyocytes chronically stimulated by isoprenaline(ISO).Methods:We used an adenoviral vector carrying the sequence for humanβ-AR(Adv.β-AR) to increase the content ofβ-AR in isolated adult rat ventricular myocytes,and examined cardiomyocyte shortening amplitude with or without ISO,after pretreatment with CGP20712A(the selectiveβ-AR antagonist),or ICI118551(the selectiveβ-AR antagonist). Results:In normal cultured myocytes,ICI118551 only inhibited basal contraction but not ISO-stimulated contraction, whereas CGP20712A suppressed the ***,in myocytes cultured with ISO,ICI118551 inhibited both basal and ISO-stimulated contraction,and so did *** ofβ-AR augmented basal contraction in myocytes cultured for 24 h,but not ISO-stimulated contraction,and the effects were abolished by ***20712A down regulated both basal and ISO-stimulated contraction in myocytes overexpressedβ-AR. Overexpression ofβ-AR augmented the basal and ISO-stimulated contraction amplitude in myocytes cultured with ISO,and the effects were abolished by CGP20712A,but not ***:The results suggested that overexpression ofβ-AR improved the contraction of myocytes injured by *** protective effect was mediated by nativeβ-*** ofβ-adrenoceptor may be a potential gene therapy approach for treating heart failure induced by a long term high sympathetic stimulation.

关键词： β2-adrenoceptor cardiomyocyte heart failure contraction

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β1- and β2-adrenergic activations enhance excitatory synaptic transmission in layerⅤ/Ⅵpyramidal neurons of the medial prefrontal cortex of rats

β1- and β2-adrenergic activations enhance excitatory synap...

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中国神经科学学会第四次会员代表大会暨第八届全国学术会议

作者： Ze-Jun FENG,Chun-Lei ZHANG,Feng YI,Xue-Han ZHANG,Bao-Ming LI~* Institute of Neurobiology,Institutes of Brain Science,Fudan University,Shanghai 200032,China

＜正＞β-adrenoceptors（β-ARs） are widely distributed throughout the central nervous *** previous work showed thatβ-AR activation facilitates excitatory synaptic transmission in pyramidal cells of the medial prefrontal cortex （mPFC）.However,little is known about theβ1- andβ2-AR regulations of synaptic transmission in the *** present study investigatedβ1- andβ2-AR modulation of glutamate synaptic transmission in layer V/VI pyramidal cells of the rat *** results show that,（1） stimulation ofβ1-ARs does not affect the frequency or the amplitude of mEPSCs,and stimulation ofβ2-ARs increases the frequency but not amplitude of mEPSCs;（2） stimulation ofβ2-ARs suppresses the paired-pulse ratio of eEPSC,and stimulation ofβ1-ARs does not;and（3） stimulations ofβ1- andβ2-ARs both enhance the amplitude of eEPSC and non-MDA-R current and NMDA-induced *** together,these results suggest that plandβ2-AR activations both facilitate excitatory synaptic transmission in mPFC pyramidal cells.

关键词： β1-adrenoceptor β2-adrenoceptor excitatory synaptic transmission medial prefrontal cortex rat

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