Th17細(xì)胞在變應(yīng)性鼻炎小鼠的表達(dá)及調(diào)控作用
發(fā)布時間：2019-03-15 08:34
【摘要】：研究目的T輔助性(Helper T, Th)細(xì)胞17是一種新的Th細(xì)胞亞型,已有研究證實(shí)它能夠通過產(chǎn)生白介素(Interleukin,IL) 17A、IL-17F以及IL-22等細(xì)胞因子,在多種自身免疫性疾病和變態(tài)炎癥反應(yīng)中發(fā)揮作用,從而引起了Th1/Th2細(xì)胞的經(jīng)典模式的調(diào)整[1]。目前,關(guān)于Th17細(xì)胞在變應(yīng)性鼻炎(Allergic Rhinitis, AR)中的研究很少,其相關(guān)的變態(tài)反應(yīng)機(jī)制尚不明確。本實(shí)驗(yàn)通過探討AR小鼠與正常小鼠、IL-17抗體干預(yù)后的AR小鼠在行為學(xué)、組織病理學(xué),Th1、Th2、Th17細(xì)胞相關(guān)免疫學(xué)細(xì)胞因子的數(shù)量以及Th17細(xì)胞特異性轉(zhuǎn)錄因子的表達(dá)水平,評價AR發(fā)病機(jī)制中Thl7細(xì)胞的可能作用及相關(guān)潛在的治療方法。 研究方法選取BALB/c小鼠30只,雌雄各半,分別隨機(jī)分為三組,對照組,模型組和干預(yù)組,每組10只。以卵清蛋白(Ovalbumin, OVA)經(jīng)典致敏法制作模型組和干預(yù)組BALB/c小鼠AR模型,造模成功后干預(yù)組予IL-17抗體治療。對照組以生理鹽水替代同方法同劑量給藥。采集外周血酶聯(lián)免疫吸附法(enzyme-linked immunosorbent assay, ELISA)檢測并分析各組小鼠IL-17、IL-4和干擾素(Interferon-γ, IFN-γ)。并取鼻粘膜行反轉(zhuǎn)錄聚合酶鏈反應(yīng)法(reverse transcriptase polymerase chain reaction, RT-PCR)測維甲酸相關(guān)孤兒核受體(Retinoic acid-related orphan receptorγt, RORγt)mRNA的表達(dá)水平及行蘇木素-伊紅(hematoxylin-eosin, HE)染色。實(shí)驗(yàn)結(jié)果以x-±s表示,采用SPSS13.0統(tǒng)計(jì)分析軟件對三組數(shù)據(jù)符合正態(tài)性分布、方差齊的進(jìn)行單因素方差分析及多組均數(shù)差別的多重比較(LSD法)。偏態(tài)或方差不齊的數(shù)據(jù)進(jìn)行多組數(shù)據(jù)的Kruskal-Wallis檢驗(yàn),及Bonferroni法兩兩比較平均秩和。以P0.05為有統(tǒng)計(jì)學(xué)意義。 研究結(jié)果模型組和干預(yù)組小鼠均激發(fā)出典型的癥狀,按評分標(biāo)準(zhǔn)造模成功。干預(yù)組經(jīng)過干預(yù)后,噴嚏明顯減少,偶有流涕,撓鼻現(xiàn)象基本消失。三組小鼠癥狀學(xué)分析模型組評分高于對照組、干預(yù)組(q值分別為10.22、9.10,P值均0.05)。模型組小鼠血清中IL-17、IL-4及鼻粘膜RORγtmRNA的表達(dá)水平明顯增高,與對照組相比差異有統(tǒng)計(jì)學(xué)意義(q/Z值分別為14.6、16.58、8.19,P值均0.05)；而IFN-γ水平明顯降低(q=8.02,P0.05)。干預(yù)組IL-17、IL-4、RORγtmRNA的表達(dá)低于模型組(q/Z=11.03、13.90、6.95,P0.05)：而IFN-Y有所升高(q=6.94,P0.05),差異均具有統(tǒng)計(jì)學(xué)意義。造模后鼻黏膜組織病理學(xué)發(fā)生炎癥性反應(yīng),纖毛中斷、柱狀細(xì)胞充盈、杯狀細(xì)胞增多,血管擴(kuò)張,腺體增生、擴(kuò)張,粘液分泌增加,嗜酸性粒細(xì)胞浸潤；干預(yù)組改善顯著。 結(jié)論在AR超敏反應(yīng)中,除了與大多數(shù)研究中一致的結(jié)果——關(guān)于AR中,Th1/Th2細(xì)胞因子平衡問的Th2細(xì)胞因子過度極化以外,Th17細(xì)胞的分化亦占優(yōu)勢,而Th1細(xì)胞受到抑制。在經(jīng)過IL-17抗體干預(yù)后,Th2、Th17細(xì)胞免疫性細(xì)胞因子數(shù)量均明顯減少,炎性反應(yīng)減退。提示這種特異性抗體治療有一定的有效性。這為變應(yīng)性鼻炎的治療學(xué)研究,提供了一個新的思路。說明抑制IL-17可以減輕變應(yīng)性鼻炎的癥狀,而針對Th17細(xì)胞特異性轉(zhuǎn)錄因子RORγt的基因干預(yù)措施更可能成為有效阻止變應(yīng)性鼻炎反應(yīng)的重要治療方法。
[Abstract]:The helper T helper (Helper T, Th) cell 17 is a new type of Th cell, and has been shown to be able to play a role in the reaction of various autoimmune diseases and perversion inflammation by producing cytokines such as interleukin (IL) 17A, IL-17F, and IL-22. So as to cause the adjustment of the classical mode of the Th1/ Th2 cells[1]. At present, the study of Th17 cells in allergic rhinitis (AR) is very low, and its associated allergic mechanism is not clear. In this experiment, the number of the related immunological cytokines and the expression level of Th17 cell-specific transcription factors in AR mice after the intervention of the IL-17 antibody in AR mice and normal mice and IL-17 antibody were discussed. To evaluate the possible effects of the Thl7 cells in the AR pathogenesis and the associated potential treatment methods. Thirty-one BALB/ c mice were randomly divided into three groups: control group, model group and intervention group. Methods: The AR model of BALB/ c mice in the model group and the intervention group were prepared by the classical sensitization method of Ovalbumin (Ovalbumin, OVA), and the model group and the intervention group were treated with IL-17 antibody treatment after the model was successfully established. In the control group, the same method and the same method were replaced with normal saline. Methods: IL-17, IL-4 and Interferon-1, IFN-1, IL-4 and Interferon-1, IFN-1 in each group were detected and analyzed by an enzyme-linked immunosorbent assay (ELISA). The expression levels of Retinoic acid-related receptor (t) mRNA and hematoxylin-eosin (HE) were measured by reverse transcription polymerase chain reaction (RT-PCR). The results of the experiment are as follows: The results of the experiment are as follows: The results of the experiment show that the data of the three groups accord with the positive distribution, the variance of the variance and the multiple comparisons (LSD) of the variance of the multiple groups of data in the three groups according to the statistical analysis software of SPSS13.0. ) the Kruskal-Wallis test of a plurality of sets of data is carried out on the data of the deviation state or the variance, and the average rank of the two comparison of the Bonferroni method And. The results of the study showed that the typical symptoms were stimulated in both the model group and the group of intervention group, and the typical symptoms were made according to the scoring criteria. The model is successful. After the intervention, the effect of the intervention group is obviously reduced, and the nasal discharge and the flexible nasal phenomenon are even. The group scores of the three groups were higher than that of the control group, and the value of the intervention group (q) was 10.22, 9.10 and P <0, respectively. In model group, the expression of IL-17, IL-4 and the expression of tmRNA in the nasal mucosa was significantly higher than that in the control group (the q/ Z value was 14.6, 16.58, 8.19, P <0.05), while the level of IFN-1 decreased significantly (q = 8.02, P0. The expression of IL-17, IL-4 and IL-4 in the intervention group was lower than that of the model group (q/ Z = 11.03, 13.90, 6.95, P0.05): while the IFN-Y increased (q = 6.94, P0.05). In order to study the significance of the study, the inflammatory reaction, the cilia interruption, the filling of the columnar cells, the increase of the goblet cells, the expansion of the vessels, the proliferation of the glands, the increase of the secretion of the mucus, the infiltration of the eosinophils, the change of the intervention group, Good results. Conclusion In AR hypersensitivity, the differentiation of Th17 cells is also dominant in addition to the excessive polarization of the Th2 cytokines in AR and Th1/ Th2 cytokine balance in AR hypersensitivity, while Th1 is thin. The number of Th2 and Th17 cellular immune cytokines was significantly reduced after the IL-17 antibody intervention. The response is reduced. It is suggested that this specific antibody will be treated with one. The efficacy of the invention. This provides a therapeutic study of allergic rhinitis. It is suggested that the suppression of IL-17 can alleviate the symptoms of allergic rhinitis, and the gene intervention measures for the specific transcription factor of Th17 cell are more likely to be more effective in the inhibition of allergic rhinitis.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R765.21
[Abstract]:The helper T helper (Helper T, Th) cell 17 is a new type of Th cell, and has been shown to be able to play a role in the reaction of various autoimmune diseases and perversion inflammation by producing cytokines such as interleukin (IL) 17A, IL-17F, and IL-22. So as to cause the adjustment of the classical mode of the Th1/ Th2 cells[1]. At present, the study of Th17 cells in allergic rhinitis (AR) is very low, and its associated allergic mechanism is not clear. In this experiment, the number of the related immunological cytokines and the expression level of Th17 cell-specific transcription factors in AR mice after the intervention of the IL-17 antibody in AR mice and normal mice and IL-17 antibody were discussed. To evaluate the possible effects of the Thl7 cells in the AR pathogenesis and the associated potential treatment methods. Thirty-one BALB/ c mice were randomly divided into three groups: control group, model group and intervention group. Methods: The AR model of BALB/ c mice in the model group and the intervention group were prepared by the classical sensitization method of Ovalbumin (Ovalbumin, OVA), and the model group and the intervention group were treated with IL-17 antibody treatment after the model was successfully established. In the control group, the same method and the same method were replaced with normal saline. Methods: IL-17, IL-4 and Interferon-1, IFN-1, IL-4 and Interferon-1, IFN-1 in each group were detected and analyzed by an enzyme-linked immunosorbent assay (ELISA). The expression levels of Retinoic acid-related receptor (t) mRNA and hematoxylin-eosin (HE) were measured by reverse transcription polymerase chain reaction (RT-PCR). The results of the experiment are as follows: The results of the experiment are as follows: The results of the experiment show that the data of the three groups accord with the positive distribution, the variance of the variance and the multiple comparisons (LSD) of the variance of the multiple groups of data in the three groups according to the statistical analysis software of SPSS13.0. ) the Kruskal-Wallis test of a plurality of sets of data is carried out on the data of the deviation state or the variance, and the average rank of the two comparison of the Bonferroni method And. The results of the study showed that the typical symptoms were stimulated in both the model group and the group of intervention group, and the typical symptoms were made according to the scoring criteria. The model is successful. After the intervention, the effect of the intervention group is obviously reduced, and the nasal discharge and the flexible nasal phenomenon are even. The group scores of the three groups were higher than that of the control group, and the value of the intervention group (q) was 10.22, 9.10 and P <0, respectively. In model group, the expression of IL-17, IL-4 and the expression of tmRNA in the nasal mucosa was significantly higher than that in the control group (the q/ Z value was 14.6, 16.58, 8.19, P <0.05), while the level of IFN-1 decreased significantly (q = 8.02, P0. The expression of IL-17, IL-4 and IL-4 in the intervention group was lower than that of the model group (q/ Z = 11.03, 13.90, 6.95, P0.05): while the IFN-Y increased (q = 6.94, P0.05). In order to study the significance of the study, the inflammatory reaction, the cilia interruption, the filling of the columnar cells, the increase of the goblet cells, the expansion of the vessels, the proliferation of the glands, the increase of the secretion of the mucus, the infiltration of the eosinophils, the change of the intervention group, Good results. Conclusion In AR hypersensitivity, the differentiation of Th17 cells is also dominant in addition to the excessive polarization of the Th2 cytokines in AR and Th1/ Th2 cytokine balance in AR hypersensitivity, while Th1 is thin. The number of Th2 and Th17 cellular immune cytokines was significantly reduced after the IL-17 antibody intervention. The response is reduced. It is suggested that this specific antibody will be treated with one. The efficacy of the invention. This provides a therapeutic study of allergic rhinitis. It is suggested that the suppression of IL-17 can alleviate the symptoms of allergic rhinitis, and the gene intervention measures for the specific transcription factor of Th17 cell are more likely to be more effective in the inhibition of allergic rhinitis.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R765.21
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