CAJ | 학술논문

目的观察白细胞介素(IL)-17、白细胞介素(IL)-4及γ干扰素(IFN-γ)在实验性自身免疫性葡萄膜炎(EAU)中的表达.方法采用健康雌性C57BL/6小鼠建立EAU模型.建模前及建模后7、14、21、28 d,取小鼠眼球作石蜡病理切片,光学显微镜观察小鼠视网膜结构变化;采用酶联免疫吸附试验检测小鼠血清中IL-17、IL-4、IFN-γ含量;采用实时荧光定量聚合酶链反应检测小鼠视网膜组织中IL-17、IL-4、IFN-γ的mRNA表达;采用蛋白质免疫印迹法检测小鼠视网膜组织中IL-17、IL-4、IFN-γ的蛋白表达.结果建模后14d,光学显微镜观察发现,葡萄膜组织结构破坏,大量炎症细胞浸润.小鼠血清中IFN-γ含量,脾及视网膜组织中IFN-γ mRNA和蛋白表达均于建模后7d达到高峰;小鼠血清中IL-4、IL-17含量,脾及视网膜组织中IL-4、IL-17 mRNA和蛋白表达均于建模后14 d达到高峰.小鼠血清中IL-17含量,脾及视网膜组织中IL-17 mRNA和蛋白表达升高程度均较IFN-γ、IL-4更为显著.建模前与建模后7、14、21 d小鼠血清中IL-17、IL-4、IFN-γ含量比较,差异均有统计学意义(F=1 817.346、268.600、164.621,P＜0.05);建模前与建模后28d小鼠血清中IL-17、IL-4、IFN-γ含量比较,差异无统计学意义(P＞0.05).建模前与建模后7、14、21d小鼠脾(F=312.67、114.250、216.220)及视网膜(F=271.504、85.370、80.722)组织中IL-17、IL-4、IFN-γ蛋白表达比较,差异有统计学意义(P＜0.05);建模前与建模后28 d小鼠脾及视网膜组织中IL-17、IL-4、IFN-γ蛋白表达比较,差异无统计学意义(P＞0.05).结论 IL-17、IL-4、IFN-γ在EAU中均明显表达,共同参与了EAU疾病的过程.
목적 관찰백세포개소(IL)-17、백세포개소(IL)-4급γ간우소(IFN-γ)재실험성자신면역성포도막염(EAU)중적표체.방법 채용건강자성C57BL/6소서건립EAU모형.건모전급건모후7、14、21、28 d,취소서안구작석사병리절편,광학현미경관찰소서시망막결구변화;채용매련면역흡부시험검측소서혈청중IL-17、IL-4、IFN-γ함량;채용실시형광정량취합매련반응검측소서시망막조직중IL-17、IL-4、IFN-γ적mRNA표체;채용단백질면역인적법검측소서시망막조직중IL-17、IL-4、IFN-γ적단백표체.결과 건모후14d,광학현미경관찰발현,포도막조직결구파배,대량염증세포침윤.소서혈청중IFN-γ함량,비급시망막조직중IFN-γ mRNA화단백표체균우건모후7d체도고봉;소서혈청중IL-4、IL-17함량,비급시망막조직중IL-4、IL-17 mRNA화단백표체균우건모후14 d체도고봉.소서혈청중IL-17함량,비급시망막조직중IL-17 mRNA화단백표체승고정도균교IFN-γ、IL-4경위현저.건모전여건모후7、14、21 d소서혈청중IL-17、IL-4、IFN-γ함량비교,차이균유통계학의의(F=1 817.346、268.600、164.621,P＜0.05);건모전여건모후28d소서혈청중IL-17、IL-4、IFN-γ함량비교,차이무통계학의의(P＞0.05).건모전여건모후7、14、21d소서비(F=312.67、114.250、216.220)급시망막(F=271.504、85.370、80.722)조직중IL-17、IL-4、IFN-γ단백표체비교,차이유통계학의의(P＜0.05);건모전여건모후28 d소서비급시망막조직중IL-17、IL-4、IFN-γ단백표체비교,차이무통계학의의(P＞0.05).결론 IL-17、IL-4、IFN-γ재EAU중균명현표체,공동삼여료EAU질병적과정.
Objective To observe the expression of interleukin (IL)-t7,IL-4 and interferon γ (IFN-γ) in experimental autoimmune uveoretinitis (EAU).Methods C57BL/6 mice were immunized with interphotoreceptor retinoid-binding protein 1-20 to induce EAU.The inflammatory reaction before and on 7,14,21,28 days after immunization were observed.The level of IL-17,IL-4 and IFN-γ in the serum were measured by enzyme-linked immune sorbent assay (ELISA).mRNA and protein expression of spleen and retina were analysed using quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot at the same time,respectively.Results The most serious inflammatory reaction occurred at the 14th day after immunization.The highest level of IFN-γ in serum,highest mRNA and protein expression of IFN-γ in spleen and retina of mice occurred at day 7 after being immunized.The highest level of IL-17,IL-4 in serum,highest mRNA and protein expression of IL-17,IL-4 in spleen and retina of mice occurred at day 14 after being immunized.The increase degree of IL-17 was more than IFN-γ and IL-4.At 7,14 and 21 days after immunization,compared with the pre-immunization,the level of IL-17,IL-4,IFN-γ in serum of mice were significantly increased (F=1 817.346,268.600,164.621; P＜0.05).There was no difference in the levels of IL-17,IL-4,IFN-γin serum of mice between pre-and 28 days after immunization (P＞0.05).At 7,14 and 21 days after immunization,compared with the pre-immunization,the protein expression of IL-17,IL-4,IFN-γ in spleen (F=312.67,114.250,216.220) and retina (F=271.504,85.370,80.722) of mice were significantly increased (P＜0.05).There was no difference in protein expression of IL-17,IL-4,IFN-γ in spleen and retina of mice between pre-and 28 days after immunization (P＞0.05).Conclusions There were IL-17,IL-4 and IFN-γ expression in EAU.IL-17,IL-4 and IFN-γplay a key role in the occurrence and development of the EAU.