刺參酸性粘多糖對誘發(fā)肝癌大鼠的免疫調(diào)節(jié)作用.pdf

1、2刺參酸性粘多糖對誘發(fā)肝癌大鼠的免疫調(diào)節(jié)作用摘要目的：通過建立誘發(fā)性大鼠肝癌動物模型，考察刺參酸性粘多糖(SJAMP)在誘發(fā)大鼠肝腫瘤過程中的作用，并從免疫功能角度研究其對肝腫瘤發(fā)生的影響。方法：將50只wistar大鼠隨機分為正常對照組(A組)、模型組(B組)、SAJMP低劑量組(175mg／Kg，C組)，SJAMP中劑量組(35mg／Kg，D組)，SJAMP高劑量組(70mg／Kg，E組)。除正常對照組(A組)外，其余各組灌胃2％o

2、DEN生理鹽水溶液以誘發(fā)肝腫瘤，SJAMP干預(yù)組(C組、D組、E組)同時灌胃給予SJAMP，飼養(yǎng)期間每周至少稱重一次，直至第16周；第16周用水合氯醛腹腔麻醉大鼠，腹主動脈取血，檢測ALT、AST等肝功指標(biāo)；無菌取脾、胸腺，立即稱重，計算脾指數(shù)和胸腺指數(shù)；分別計數(shù)肝臟上有一條直徑大于3mm和5mm的瘤結(jié)節(jié)，并測量最大瘤結(jié)節(jié)的長徑和短徑，計算抑瘤率；貼壁法分離純化M巾，用中性紅吞噬實驗檢測脾巨噬細胞吞噬功能；MTT法檢測脾巨噬細胞的殺傷功

3、能；MTT法檢測自然殺傷細胞活性：ELISA法檢測血清中TNF—a和IL一2的水平；流式細胞術(shù)分析大鼠外周血T淋巴細胞亞群。結(jié)果：①與正常對照組相比，誘癌實驗組大鼠體重增長較慢，至實驗第16周時，E組和B組大鼠體重有統(tǒng)計學(xué)差異(P005)；②誘癌實驗組ALT、AST和GGT均升高(P005)，其中模型組升高最為顯著，隨干預(yù)劑量增加，SJAMP干預(yù)組以上指標(biāo)呈逐漸下降趨勢，且與模型組比較有統(tǒng)計學(xué)意義(P001)；③模型組胸腺指數(shù)與正常組組

4、相比有所降低(P005)，但脾指數(shù)有所升高(P005)，與模型組相比，SJAMP干預(yù)組脾指數(shù)和胸腺指數(shù)明顯升高(PO05)，并呈現(xiàn)出劑量一效應(yīng)關(guān)系；④SJAMP可顯著抑制肝腫瘤的發(fā)生，減少癌結(jié)節(jié)的發(fā)生數(shù)量，SJAMP干預(yù)組大鼠大于3mm和大于5mm的結(jié)節(jié)數(shù)明顯少于模型組(P005)；最大結(jié)節(jié)的平均體積明顯小于模型組(P001)；⑤模型組和S2wP干預(yù)組巨噬細胞吞噬能力較正常對照組均增(P005)，SJAMP干預(yù)組增加更為明顯，其中D組和

5、E組與C組相比較，有統(tǒng)計學(xué)意義(P001)；⑥與正常對照組相比，模型組巨噬細胞的殺傷能力降低(PO01)，SJAMP干預(yù)組巨噬細胞的殺傷能力較模型組增加(P001)；⑦模型組自然殺傷細胞活性較正常對照組明顯降低(P001)，SJAMP干預(yù)可以提高誘癌大鼠的自然殺傷細胞活性(PO05)；⑧模型組和SJAMP干預(yù)組的血清TNF—a水平高于正常組(P001)，提示誘癌各組大鼠均可能存在肝臟損傷，SJAMP干預(yù)組中D組和E組與模型組相比較，差別

6、均具有統(tǒng)計學(xué)意義(P001)；⑨模型組大鼠血清IL2水平明顯降低于正常組(P001)，SJAMP干預(yù)組的IL一2TheEffectofStichopusJaponicusAcidMucopolysaccharideontheDevelopmentandImmuneFunctionofHepatocellularCarcinomaRatInducedbyDiethylnitrosamineAbstractObjectiveThrought

7、heestablishmentofratlivercancermodel，theroleofStichopusjaponicusacidmucopolysaccharide(SJAMP)inthedevelopmentofhepatocellularcarcinomainducedbydiethylnitrosamine(DEN)wasexploredinratsandthepossibleimmunologicalmechanisms

8、wasdeterminedMethods50wistarratswererandomlydividedintofivegroups：normalcontrolgroup(groupA)，modelgroup(groupB)，SAJMPlowdosegroup(175mg／Kg，groupC)，SJAMPmoderatedosegroup(35mg／Kg，groupD)，SJAMPhighdosegroup(70mg／Kg，groupE)

9、byweightInadditiontonormalcontrolgroup，alltheratsweregivenorally2‰DE]Nsalinesolutiontothe16thweektoinducehepatocelluarcarcinomameanwhiletheywereintervenedwimdifferentdosesofSJAMPduringtheprocessofcarcinogenesisAlltherats

10、wereweighedonceaweekduringthebreeding，andattheendofthe16thweek，a11theratswereanaesthetizedbyusingchloralhydratebloodsamplesweredrewfromtheabdominalaorta；spleenandthymuswereexcisedundersterileconditionsandweighed，spleenin

11、dexandthymusindexwerecalculatedThenumberofthenodulesmorethan3ramor5mmindiameterwerecounted，inadditionthesizeofthelargestnodulewerecomparedamonggroupsThetumorinhibitoryrateWascalculatedMacrophagocytes(Mf)werepurifiedviaat

12、tachmentculture，macrophagephagocytosisWasobservedwithphagocytosingneutralredmethod；spleenmacrophagekillingcapabilityandNK(Naturalkiller)cellactivityweremeasuredbyMTTmethod，thelevelsofserumTNFaandIL2weredetectedwithenzyme

13、linkedimmunosorbantassay(ELISA)；TlymphocytesubsetsinperipheralbloodwereanalyzedbyflowcytometryReSultsOComparedwithnormalcontrolgroup，theinducedhepaticcarcinomaratsgainweightsloweratthe16thweek，theweightsofratsofgroupEwas

14、obviouslyincreasedwhencomparedtomodelgroupB(PO05)；②TheserumALTAST，andGGToftheinducedhepaticcarcinomarats，especiallyinmodelgroup，increasedsignificantlywhencomparedtothoseofthenormalcontrolgroup，withtheincreaseoftheSJAMPin

15、terventiondose，theaboveindicatorsdecreasedgraduallyandthereweresignificantdifferencescomparedwithmodelgroup；③Thethymusindexofmodelgroupdecreased(PO05)，whilethespleenindexincreased(P005)whencomparedtonormalgroup；withtheim