光變應(yīng)原檢測動物模型的建立以及藥物干預(yù)光變態(tài)反應(yīng)的研究.pdf

1、光變應(yīng)原檢測動物模型的建立及藥物干預(yù)光變態(tài)反應(yīng)的實(shí)驗(yàn)和臨床研究(摘要)為了建立檢測光變應(yīng)原的動物模型，本文采用小鼠耳腫脹模型分別用于接觸性和系統(tǒng)性光變應(yīng)原的檢測；并在國內(nèi)首次建立了小鼠局部淋巴結(jié)檢測結(jié)合耳腫脹程度評價的方法檢測接觸性光變應(yīng)原。在接觸性光變態(tài)反應(yīng)小鼠耳腫脹模型中，使用環(huán)磷酰胺作為免疫佐劑，在誘導(dǎo)和激發(fā)過程中同時予UVA和UVB照射，并且設(shè)立了光毒反應(yīng)組、接觸性變態(tài)反應(yīng)組和溶媒輻射組進(jìn)行對照。f結(jié)果顯示，受試物6甲基香豆索僅

2、在光變態(tài)反應(yīng)組中予激發(fā)后24小時出現(xiàn)耳腫脹庋應(yīng)，耳組織學(xué)改變符合遲發(fā)型變態(tài)反應(yīng)(DTH)的病理特征，在其余三個對照組中均不出現(xiàn)耳腫脹反應(yīng)，證實(shí)為接觸性光變應(yīng)原。在小鼠局部淋巴結(jié)檢測中結(jié)合耳腫脹程度評價，即用受試物加UVA照射連續(xù)3天在小鼠耳背部進(jìn)行激發(fā)，最后一次照射結(jié)束后24小時切取引流的耳淋巴結(jié)，用淋巴細(xì)胞計數(shù)和淋巴細(xì)胞增殖率反映耳淋巴結(jié)的增殖情況；并同時測量小鼠耳腫度以反映皮膚的刺激反應(yīng)。用MTT比色法取代[aHITdR摻入法測定淋

3、巴細(xì)胞增殖率，避免了放射性污染。結(jié)果顯示，四氯水楊酰苯胺引起局部淋巴結(jié)增殖而不伴耳腫脹反應(yīng)，為光變應(yīng)原；8一甲氧補(bǔ)骨脂素同時引起局部淋巴結(jié)增殖和耳腫脹反應(yīng)，為光毒物質(zhì)；6甲基香豆素則既無局部淋巴結(jié)增殖又無耳腫脹反應(yīng)。本方法與接觸性光變態(tài)反應(yīng)小鼠耳腫脹模型相比，優(yōu)點(diǎn)在于可迅速鑒別較強(qiáng)的光變應(yīng)原和光毒物質(zhì)，但對識別較弱的光變應(yīng)原敏感性不足。進(jìn)一步測定局部淋巴結(jié)中細(xì)胞因子的分泌情況，結(jié)果顯示，預(yù)先致敏后再進(jìn)行激發(fā)，局部淋巴結(jié)中IL4的分泌量明

4、顯升高是光變應(yīng)原區(qū)別于光毒物質(zhì)的突出特點(diǎn)，有助于兩者間的鑒別，但仍無法識別較弱的光變應(yīng)原。在系統(tǒng)性光變態(tài)反應(yīng)小鼠耳腫脹模型中，使用環(huán)磷酰胺和福氏完全佐劑作為免疫佐劑，同時設(shè)立光毒反應(yīng)組和對照組。結(jié)果顯示，受試物對氨基苯磺酰胺僅在光變態(tài)反應(yīng)組中于激發(fā)后24小時出現(xiàn)耳腫脹反應(yīng)，耳組織學(xué)改變符合DTH的病理特征，在其余兩個對照組中均不出現(xiàn)耳腫脹反應(yīng)，證實(shí)為光變應(yīng)性藥物。利用已建立的接觸性和系統(tǒng)性光變態(tài)反應(yīng)小鼠耳腫脹模型，分別用羥基氯喹、雷公藤

5、這二種藥物進(jìn)行干預(yù)，結(jié)果顯示兩藥均可抑制兩種小鼠模型的耳腫脹反應(yīng)，減輕組織炎癥反應(yīng)；進(jìn)一步研究這二種藥物對接觸性光變態(tài)反應(yīng)小鼠模型的局部淋巴結(jié)中淋巴細(xì)胞增殖和細(xì)胞因子分泌情況的影響，結(jié)果顯示，兩藥均可引起淋巴結(jié)PhotoallergicReaction：theEstablishmentofAnimalModelsandStudiesontheHydroxychioroquineandTripterygiumwilfordiiTreatm

6、ent他bstract)Inordertoestablishanimalmodelsforthedetectionofphotoallergens，murineeat“swellingmodelwasemployedtoidentifycontactandsystemicphotoallergens，andamurinclymphnodeassay(LLNA)combinedwithmeasurementofprimaryearswel

7、lingwasnewlyestablishedtoinvestigatecontactphotoallergensInthecontactphotoallergicmurineearswellingmodel，cyclophosphamidewasusedasimmunoadjuvant，whilebothUVAandUVBirradiationwereemployedintheinductionandchallengephasePho

8、totoxicity，contactallergyandvehicle／radiationcontrolgroupswerealsoprovidedatthesametimeTheresultsshowedthatmiceinthephotoallergygrouppretreatedwith6methylcoumarinandUVirradiationdevelopedevidentearswellingat24haRerchalle

9、ngeandthehistopathologyWascharacteristicofadelayedtypehypersensitivity(DTH)，whilecontrolmiceshowedneaction，indicatingthat6methylcoumarinisakindofcontactphotoallergenInthisstudy，theroutineLLNAwascombinedwithprimaryearswel

10、lingmeasurementBriefly，testchemicalswereappliedtothedorsumofbothearsfollowedbyUVAirradiationonthreeconsecutivedaysTheauricularlymphnodesdrainingthecartissuewereexcised24hfollowingthelastirradiationLymphnodecellcountsandl

11、ymphocyteproliferativeratesweredetermined，indicatinglymphnodehyperplasia；whileearswellingwasassessedindicatingskinirritatingreactionsMTTcolorimetrywasusedinsteadof【3H]thymidineincorporationinlymphocyteproliferativerateme

12、asurementtoavoidradioactivityTheresultshowedthattetrachlorosalicylanilide，as廿ongphotoallergen，causedlymphnodehyperplasiaintheabsenceofearswelling，and8methoxypsoralen，aphototoxicsubstance，inducedlymphnodehyperplasiatogeth

13、erwithearswellingbutthemildphotoallergen6methylcoumarinshowedneitherlymphnodeactivationnorearswellingreactionsComparedwithcontactphotoallergiemurineearswellingmodel，LLNAcombinedwiththemeasurementofprimaryearswellingmakes

14、itpossibletodifferentiates仃ongphotoallergensfromphototoxiesubstancequickly，butisnotsensitiveenoughtodetectmildphotoallergensFurthermore，Weanalyzedcytokineprofilesinthedraininglymphnodeandrevealedthatphotoallergencauseda仃