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AstragalosideⅣPrevents the Mitochondrial Permeability Transition Pore Opening by Inhibiting Glycogen Synthase Kinase 3β via the NO/cGMP/PKG Pathway in Cardiac Cells

AstragalosideⅣPrevents the Mitochondrial Permeability Trans...

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中国生理学会心血管生理学术研讨会

作者： yonggui he~(1,2),yuxi xie~2,huan zheng~2,yidong zhang~2,jing guo~2,jinkun xi~(2*) 1.department of internal medicine 2.heart Institute,hebei United University Tangshan 063000

Aim:AstragalosideⅣ,a major ingredient of traditional Chinese herb Radix Astragali,has been widely used for treatments of cardiovascular diseases in China,but the exact cellular and molecular mechanisms underlying its action remain *** aimed to test if astragalosideⅣmodulates mitochondrial permeability transition pore(mPTP) opening through glycogen synthase kinase-3β(GSK-3β) in H9c2 cardiac *** more,the molecular mechanism was studied,by which Astragaloside IV inhibits GSK-3βactivity in this ***:Rat heart tissue-derived H9c2 cells were purchased from *** cardiac H9c2 cells were exposed to HO(600μM) for 20 min to induce mitochondrial oxidant *** dyes including tetramethylrhodamine ethyl ester(TMRE,red) and 4-amino-5-methylamino-2’, 7’-difluorofluorescein(DAF-FM,green) diacetate were used to image mitochondrial membrane potential(Δψm) and nitric oxide(NO),*** images were obtained with laser-scanning confocal *** red fluorescence was excited with a 543 nm line of he-Ne laser line and imaged through a 560 nm long-path *** green fluorescence was excited at 488 nm and imaged through a 525 nm long-path *** cell viability was assessed by propidium iodide fluorometry using a fluorescence *** in 12-well plates coated with laminin were incubated in standard Tyrode solution,and fluorescence intensity was measured at the excitation and emission wavelengths of 540 and 590 nm,respectively.H9c2 cells were exposed to astragalosideⅣfor 20 min,the GSK-3β(Ser),Akt(Ser),and VASP(Ser) activities were determined by measuring their phosphorylation statuses with Western blot. Results:Cardiac cells treated with different dose of astragalosideⅣ(30μM-80μM),and 50μM astragalosideⅣsignificantly increased GSK-3βphosphorylation at Ser(121.1±2.7%) and prevented oxidative stress-induced loss of mitochondrial membrane potential(MMP) in H9c2 cardiac cells(89.7±4.3%vs.49.7±6.3%in control).The GSK-3βin

关键词： AstragalosideⅣ mPTP GSK-3β cGMP/PKG NO PI3K/Akt

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