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1、櫛桿菌多糖硫酸酯化及酯化前后生物活性比較探究摘要:目的以櫛桿菌多糖(APD)為研究對象,進(jìn)行硫酸酯化(SAPD),并測定其抗腫瘤活性和免疫調(diào)節(jié)活性。方法APD采用氯磺酸-吡啶法進(jìn)行酯化,溴化二苯偶氮鹽(MTT)比色法研究樣品在體外對腫瘤細(xì)胞株的抑制作用和促進(jìn)正常小鼠脾臟細(xì)胞增生的作用。結(jié)果紅外光譜證明SAPD已經(jīng)形成硫酸酯,其取代度為0.918。APD對腫瘤細(xì)胞株幾乎無細(xì)胞毒性,SAPD對A549肺癌細(xì)胞株的抑制率為31%。APD對ConA誘導(dǎo)的脾臟細(xì)胞增生具有促進(jìn)作用,但對LPS誘導(dǎo)的脾臟細(xì)胞增生無促進(jìn)作用。SAPD低
2、濃度時(shí)對ConA誘導(dǎo)的脾臟細(xì)胞增生有促進(jìn)作用,對LPS誘導(dǎo)的脾臟細(xì)胞增生無促進(jìn)作用。結(jié)論APD經(jīng)過硫酸酯化后抗腫瘤活性顯著增強(qiáng),對ConA誘導(dǎo)的脾臟細(xì)胞增生有一定的促進(jìn)作用。關(guān)鍵詞:櫛桿菌多糖;硫酸酯化;抗腫瘤;免疫調(diào)節(jié)中圖分類號:Q539文獻(xiàn)標(biāo)識碼:A文章編號:1672-979X(2007)01-0001-04SulfationofPolysaccharidefromArthrobactersp.andComparativeStudyonBioactivityofPolysaccharideandItsSulfated
3、DerivativeWANGFeng-qin,GENGYue10(CollegeofLifeScience,ShandongNormalUniversity,Jinan250014,China)Abstract:ObjectiveTostudythesulfationofthepolysaccharidefromArthrobactersp.(APD)anddeterminetheantitumorandimmunitymodulationactivitiesofitanditssulfatedderivative(SA
4、PD).MethodsAPDwaschemicallymodifiedbychlorosulfonicacid-pyridinemethod.MTTcolourimetrywasusedtoevaluatetheantitumoractivityagainsthumantumorcellsinvitro,aswellastheproliferation-enhancingeffectonspleenlymphocytesofnormalmice.ResultsTheIRspectrumshowedthatSAPDhadb
5、eenformed,thesubstituteddegreewas0.918.APDhadlittlecytotoxicactivitytowardsthecancercells,whileSAPDinhibitedthegrowthofA549lungcancercellsremarkablywith31%inhibitoryratio.ThoughAPDhadnoeffectontheproliferationofLPS-inducedspleencells,itcouldenhancetheproliferatio
6、nofConA-inducedspleencells.SAPDwithlowdosecouldpromotetheproliferationofConA-inducedspleencells,butalsohadnoeffectontheproliferationof10LPS-inducedspleencells.ConclusionTheantitumoractivityofAPDisremarkablyenhancedafteritwassulfated.APDcanalsopromotetheproliferat
7、ionofConA-inducedspleencells.Keywords:polysaccharidefromArthrobactersp;sulfation;antitumor;immunitymodulation多糖的硫酸酯化是多糖化學(xué)修飾研究中的一個(gè)熱點(diǎn),實(shí)驗(yàn)表明硫酸酯化多糖具有抗病毒、抗腫瘤、抗凝血等作用。本實(shí)驗(yàn)對櫛桿菌中的多糖進(jìn)行分離純化,并對其中一種組分(APD)進(jìn)行硫酸酯化,得到硫酸酯化多糖(SAPD),然后對APD和SAPD的抗腫瘤活性和免疫活性進(jìn)行了初步研究。1材料1.1APD櫛桿菌(Arthroba
8、ctersp.)發(fā)酵產(chǎn)物分離到的多糖,去蛋白質(zhì)后經(jīng)20%,40%,60%,80%乙醇分級沉淀得到多糖組分,選取80%乙醇沉淀(APD)作為實(shí)驗(yàn)材料。1.2試劑吡啶(山東化學(xué)研究所);氯磺酸(中國醫(yī)藥上?;瘜W(xué)試劑廠);明膠(上?;瘜W(xué)試劑公司);無水甲酰胺(天津廣成化學(xué)試劑公司);硫酸鉀、氯化鋇(天津天河化工廠);RPM