資源描述:
《l.casei zhang pg對小鼠巨噬細(xì)胞吞噬功能的影響》由會(huì)員上傳分享,免費(fèi)在線閱讀,更多相關(guān)內(nèi)容在應(yīng)用文檔-天天文庫。
1、L.caseiZhangPG對小鼠巨噬細(xì)胞吞噬功能的影響【摘要】目的觀察干酪乳桿菌(L.caseiZhang)肽聚糖(peptidoglycan,PG)對小鼠腹腔巨噬細(xì)胞吞噬率、吞噬指數(shù)及巨噬細(xì)胞表面C3b受體的影響。方法應(yīng)用酵母菌體外吞噬法觀察巨噬細(xì)胞的吞噬功能;應(yīng)用伊紅染色法計(jì)數(shù)酵母細(xì)胞花環(huán)(Yc-花環(huán))形成率;應(yīng)用動(dòng)物實(shí)驗(yàn)法進(jìn)行小鼠毒性試驗(yàn)。結(jié)果L.caseiZhangPG組腹腔巨噬細(xì)胞吞噬率和吞噬指數(shù)分別為67.69±7.08和2.11±0.39,明顯高于對照組的31.22±4.18和
2、0.78±0.14,差異有顯著性。L.caseiZhangPG組巨噬細(xì)胞表面Yc-花環(huán)形成率為60.01±4.02,明顯高于對照組的35.07±5.71,差異有顯著性。毒性試驗(yàn)結(jié)果顯示,連續(xù)觀察一周L.caseiZhangPG組小鼠,小鼠均無聳毛、消瘦或食欲不振等中毒現(xiàn)象。結(jié)論L.caseiZhangPG對巨噬細(xì)胞有一定的激活作用,能提高其吞噬功能,且安全可靠?!娟P(guān)鍵詞】干酪乳桿菌肽聚糖巨噬細(xì)胞C3b受體 EFFECTOFL.CASEIZHANGPEPIDOGLYCANONPHAGOCYTOS
3、ISOFMICROPHAGEINMICE ShuangJie1,YongSheng1,ZhangQijin2 (1.Dept.ofpathogenbiology,QinghaiUniversityMedicalSchool; 2.CollegeofVeterinaryScienceandMedicine,InnerMongoliaAgriculturalUniversity) AbstractObjective ToinvestigatetheeffectsofL.caseiZhangpeptidoglycanontheph
4、agocytosisrate,phagocytosisindexandsurfaceC3breceptorofmicrophageinmice.MethodsThephagocyticfunctionofmicrophageingratealice.ResultsThephagocytosisrateandindexofperitonealmicrophagestimulatedbyL.caseiZhangpeptidoglycann;7.08and2.11±0.39,respectivelyandsignificant
5、lyhigherthanthoseincontrol(31.22±4.18and0.78±0.14).TheYcrosetteformationinsurfaceofmacrophageinL.caseiZhangpeptidoglycangroupn;4.02,significantlyhigherthanthatofcontrolgroup(35.07±5.71),andshoaciatedandhavingadelicateappetite,etc.andallofthemsurvive
6、d.ConclusionL.caseiZhangpeptidoglycancanactivatethemicrophing.ageandpromotethefunctionofitsphagocytosisandsafeforus. Key,10min)收集菌體;將菌體懸浮于蒸餾水中,超聲處理(20min,3次),上清液差速離心(1000rpm,15min;10000rpm,20min)收集粗細(xì)胞壁;取適量粗細(xì)胞壁加SDS溶液離心(10000rpm,20min),收集粗肽聚糖;將粗肽聚糖洗凈后溶于胰蛋白酶緩沖液(pH7.5)中,37
7、℃水浴振蕩20h,離心(10000rpm,20min)獲得酶處理粗細(xì)胞壁,粗細(xì)胞壁用無水乙醇脫水;最后懸浮在三氯乙酸溶液(10g/dL)中去除共價(jià)結(jié)合的磷壁酸,離心(10000rpm,20min)后將沉淀物用蒸餾水洗滌、用無水乙醇脫水干燥,保存。具體步驟見 1.2.4.5Yc-花環(huán)試驗(yàn)方法:將Yc懸液滴加于單層巨噬細(xì)胞上,平置(4℃)1h,繼用緩沖液沖洗玻片,至無酵母顆粒沖下為止。加2%戊二醛固定、HE染色后,用顯微鏡計(jì)數(shù)細(xì)胞。以一個(gè)巨噬細(xì)胞周圍附有6個(gè)或6個(gè)以上酵母菌者計(jì)作一個(gè)花環(huán),隨機(jī)觀察200個(gè)巨噬細(xì)胞,計(jì)算花環(huán)形成率?! ?
8、.2.5實(shí)驗(yàn)組小鼠急性毒性試驗(yàn) 以每只1、2、4mg三個(gè)梯度劑量的L.caseiZhangPG對小鼠行腹腔注射,每個(gè)劑量組注射6只小鼠,觀察1周?! ?.2.6統(tǒng)計(jì)學(xué)處理方法 采用SPSS11.0軟件對各試驗(yàn)數(shù)據(jù)進(jìn)行