清開(kāi)靈眼用凝膠對(duì)實(shí)驗(yàn)性自身免疫性葡萄膜炎大鼠Th1、Th2細(xì)胞的影響的實(shí)驗(yàn)研究.pdf

1、提要目的：目的：利用光感受器間維生素A類結(jié)合蛋白(interphoteceptretinoidbindingproteinIRBP)的致葡萄膜視網(wǎng)膜炎活性建立實(shí)驗(yàn)性自身免疫性葡萄膜炎（ExperimentalAutoimmuneUveitisEAU）模型，應(yīng)用清開(kāi)靈眼用凝膠對(duì)其干預(yù)，探討清開(kāi)靈眼用凝膠治療實(shí)驗(yàn)性自身免疫性葡萄膜炎大鼠的可行性及對(duì)其Th1、Th2細(xì)胞的影響方法：方法：1、觀察清開(kāi)靈眼用凝膠對(duì)EAU大鼠的作用并與地塞米松點(diǎn)眼

2、組進(jìn)行比較，觀察其療效，為其進(jìn)一步應(yīng)用于臨床葡萄膜炎的防治奠定基礎(chǔ)。將健康無(wú)眼病的清潔級(jí)雌性Lewis大鼠隨機(jī)分為三組，每組36只均將200μl含有100μgIRBP11771191及完全弗氏佐劑的混合液，充分乳化后在Lewis大鼠雙側(cè)后足墊皮下、尾根部及軀干皮下均勻注射6點(diǎn)進(jìn)行免疫建立動(dòng)物模型。A組為模型對(duì)照組，免疫后均無(wú)藥物點(diǎn)眼，B組為清開(kāi)靈眼用凝膠點(diǎn)眼組，免疫后第1天起予清開(kāi)靈眼用凝膠點(diǎn)眼，3次日，C組為地塞米松點(diǎn)眼組，免疫后第1

3、天起予0.1%地塞米松點(diǎn)眼，3次日。免疫后第1天，3天，5天，7天，9天，11天，13天，15天，17天對(duì)動(dòng)物模型應(yīng)用裂隙燈觀察眼前節(jié)炎癥反應(yīng)，記錄發(fā)病程度，進(jìn)行炎癥分級(jí)并于第9天處死大鼠36只組，行眼部病理檢查，進(jìn)行病理分級(jí)。2、清開(kāi)靈眼用凝膠對(duì)實(shí)驗(yàn)性自身免疫性葡萄膜炎大鼠Th1、Th2細(xì)胞的影響于免疫后第9天過(guò)量麻醉處死大鼠，制備T細(xì)胞的單細(xì)胞懸液。應(yīng)用流式細(xì)胞儀分析各組CD4CD8IFNγ的百分比及計(jì)算CD4CD8的比值。應(yīng)用電泳

4、儀分析各組0，3579天外周血中IFNγ，IL4細(xì)胞因子的表達(dá)強(qiáng)弱。應(yīng)用實(shí)時(shí)熒光定量PCR分析各組0，3579天淋巴結(jié)中IFNγ，IL4細(xì)胞因子的表達(dá)強(qiáng)弱。應(yīng)用酶聯(lián)免疫吸附測(cè)定法（Elisa）測(cè)定各組大鼠脾臟和淋巴結(jié)細(xì)胞培養(yǎng)上清及房水中IFNγ，IL4的濃度。結(jié)果：結(jié)果：1.清開(kāi)靈眼用凝膠對(duì)EAU大鼠的療效觀察。（1）從大鼠眼部炎癥表現(xiàn)所見(jiàn)，清開(kāi)靈眼用凝膠和地塞米松注射液均抑制EAU大鼠的發(fā)病，并且兩者之間差別不大。各組大鼠眼部的炎癥得

5、分分別為：模型對(duì)照組：(3.600.52)分。清開(kāi)靈組：(1.20.63)分。地塞米松組：（1.00.67）分。（2）病理檢查（HE染色）所見(jiàn)，模型對(duì)照組大鼠眼球外層視網(wǎng)膜廣泛而重度的AnexperimentalresearchontheinfluenceofQingKaiLingOphthalmicGeltoTh1Th2cellsinratswithexperimentalautoimmuneuveitisSpeciality:Oph

6、thalmologyCombinationofTCMWesternMedicineAuth:WeiHhaSuTut:Prof.BiHongshengABSTRACTObjective:ExpletheimpactoftheQingKaiLingOphthalmicGelonTh1Th2cellsintheratswithexperimentalautoimmuneuveitis(EAU)providethetheeticalbasisf

7、clinicalwidelyusingMethods:1.ToobservetheeffectofQingKaiLingOphthalmicGelontheratswithexperimentalautoimmuneuveitis(EAU)compareditseffectwiththedexamethasoneinjectionthroughtheintraperitonealinjectionappraisingiteffectfi

8、tsfurtherappliedtoclinicaluveitislaythefoundationfthepreventioncontrol.EAUwasinducedinLewisratsbyimmunizationwithIRBP1177–1191incompleteFreund’sadjuvant.Theratswereromlydividedintothreegroups.GroupA(modelcontrol)groupB(Q

9、ingKaiLingOphthalmicGelpointingeye)groupC(dexamethasoneinjectionthroughtheintraperitonealinjection)evaluatedclinicallyondays07911131517evaluatedinflammationpathologicallyondays9.2.DiscusstheimpactoftheQingKaiLingOphthalm

10、icGelonTh1Th2cellsintheratswithexperimentalautoimmuneuveitis(EAU)EAUwasinducedinLewisratsbyimmunizationwithIRBP1177–1191incompleteFreund’sadjuvant.Theratswereromlydividedintothreegroups.GroupA(modelcontrol)groupB(QingKai

11、LingOphthalmicGelpointingeye)groupC(dexamethasoneinjectionthroughtheintraperitonealinjection).Executedratsondays9lymphocytesfromlymphnodes(LNs)spleenweresubjectedculturedthatstimulatedbyIRBP1177–1191toflowcytometrytoanal