简介:目的:探讨银杏内酯B对谷氨酸诱导的人神经母细胞瘤(SH-SY5Y)细胞凋亡的保护作用及其机制。方法:用2mmol/L谷氨酸诱导SH-SY5Y细胞凋亡建立模型,以不同浓度的银杏内酯B(10、50和250μmol/L)进行保护,MTT比色法检测细胞存活率;Hoechst33258染色法观察药物作用后细胞凋亡的形态学变化;流式细胞仪检测细胞凋亡率;Fura-2/AM双波长荧光法测定细胞内钙离子浓度;Westernblot检测线粒体和胞浆中CytC蛋白的表达。结果:2mmol/L谷氨酸作用SH-SY5Y细胞24h后细胞存活率为0.721±0.013,与正常组相比明显降低;细胞核固缩裂解,呈凋亡特征性改变;细胞凋亡率为(13.27±0.08)%、细胞内钙离子浓度为359±11nmol/L,均明显升高。与凋亡组相比,银杏内酯B(10、50、250)μmol/L保护组细胞存活率分别为0.768±0.014、0.819±0.013、0.897±0.010,均明显升高;细胞核核固缩裂解减少,形态有所改善;细胞凋亡率分别为11.33±0.57、7.02±0.47、5.85±0.24;细胞内钙离子浓度为302±9nmol/L、243±9nmol/L、186±8nmol/L,均明显降低;Westernblot结果显示,与凋亡组相比银杏内酯B保护组可以显著减少细胞线粒体以及胞浆中CytC蛋白的含量。结论:银杏内酯B对谷氨酸诱导的SH-SY5Y细胞凋亡具有保护作用,其机制可能与银杏内酯B降低了细胞内钙离子的浓度及阻断了CytC从线粒体释放入胞浆中有关。
简介:Houttuyniacordatapolysaccharide(HCP)isextractedfromHouttuyniacordata,akeytraditionalChinesemedicine.ThestudywastoinvestigatetheeffectsofHCPonintestinalbarrierandmicrobiotainH1N1virusinfectedmice.MicewereinfectedwithH1N1virusandorallyadministratedHCPatadosageof40mg(kg^-1(d^-1.H1N1infectioncausedpulmonaryandintestinalinjuryandgutmicrobiotaimbalance.HCPsignificantlysuppressedtheexpressionofhypoxiainduciblefactor-1αanddecreasedmucosubstancesingobletcells,butrestoredthelevelofzonulaoccludens-1inintestine.HCPalsoreversedthecompositionchangeofintestinalmicrobiotacausedbyH1N1infection,withsignificantlyreducedrelativeabundancesofVibrioandBacillus,thepathogenicbacterialgenera.Furthermore,HCPrebalancedthegutmicrobiotaandrestoredtheintestinalhomeostasistosomedegree.TheinhibitionofinflammationwasassociatedwiththereducedlevelofToll-likereceptorsandinterleukin-1βinintestine,aswellastheincreasedproductionofinterleukin-10.OraladministrationofHCPalleviatedlunginjuryandintestinaldysfunctioncausedbyH1N1infection.HCPmaygainsystemictreatmentbylocalactingonintestineandmicrobiota.Thisstudyprovedthehigh-valueapplicationofHCP.
简介:Thepresentstudywasdesignedtoinvestigatetheanti-sepsiseffectsofphyscion8-O-β-glucopyranoside(POG)isolatedfromRumexjaponicasandexploreitspossiblepharmacologicalmechanisms.POGwasextractedfromR.japonicasbybioactivity-guidedisolationwiththeanti-sepsisagents.SurvivalanalysisinsepticmouseinducedbyLPSandheat-killedEscherichiacoliwereusedtoevaluatetheprotectiveeffectofPOG(40mg·kg~(-1),i.p.)onsepsis.CytokinesincludingTNF-α,IL-1βandIL-6inRAW264.7cellsinducedbyLPS(100ng·mL~(-1))weredeterminedbyELISA.Inaddition,theproteinsexpressionsofTLR2andTLR4weredeterminedbyWesternblottingassay.OurresultsdemonstratedthatPOG(40mg·kg~(-1),i.p.)possessedsignificantprotectiveactivityontheendotoxemicmice.ThePOGtreatment(20,40,and80μg·mL~(-1))significantlydecreasedtheTNF-α,IL-1βandIL-6inducedbyLPS(P<0.01)inaconcentration-dependentmanner.Furthermore,theTLR4andTLR2proteinswerealsodown-regulatedbyPOGat20(P<0.01),40(P<0.01),and80μg·mL~(-1)(P<0.01).ThepresentstudydemonstratedthatthePOGextractedfromR.japonicaspossessedsignificantanti-sepsiseffectonendotoxemicmice,andcanbedevelopedasanoveldrugfortreatingsepsisinthefuture.
简介:目的:建立同时测定不同产地半枝莲中野黄芩苷、黄芩苷、野黄芩素、异野黄芩素、木犀草素、芹菜素、黄芩素和汉黄芩素含量的高效液相色谱法。方法:采用AgilentZORBAXSB-C18色谱柱(250mm×4.6mm,5μm),以乙腈-0.1%甲酸水为流动相,梯度洗脱,检测波长:350nm,流速:1.0mL·min^-1,柱温:25℃。结果:半枝莲药材中野黄芩苷、黄芩苷、野黄芩素、异野黄芩素、木犀草素、芹菜素、黄芩素和汉黄芩素分别在7.23~72.32、3.28~32.76、2.35~23.53、2.54~25.37、1.77~17.65、2.26~22.57、1.95~19.48、3.52~35.24μg呈良好线性关系,相关系数r在0.9983~0.9999,平均加样回收率为98.9%~99.9%。结论:该方法简单、快速,为半枝莲的质量控制提供了参考。
简介:Inthepresentstudy,aseriesofnovelnitricoxide-hydrogensulfidereleasingderivativesof(S)-3-n-butylphthalide((S)-NBP)weredesigned,synthesized,andevaluatedaspotentialantiplateletagents.CompoundNOSH-NBP-5displayedthestrongestactivityininhibitingthearachidonicacid(AA)-andadenosinediphosphate(ADP)-inducedplateletaggregationinvitro,with3.8-and7.0-foldmoreeffectivenessthan(S)-NBP,respectively.Furthermore,NOSH-NBP-5couldreleasemoderatelevelsofNOandH2S,whichwouldbebeneficialinimprovingcardiovascularandcerebralcirculation.Moreover,NOSH-NBP-5couldrelease(S)-NBPwhenincubatedwithratbrainhomogenate.Inconclusion,thesefindingsmayprovidenewinsightsintothedevelopmentofnovelantiplateletagentsforthetreatmentofthrombosis-relatedischemicstroke.