CAJ | 학술논문

背景雷帕霉素不仅具有抗菌作用,而且是一种良好的免疫抑制剂,可用于多种自身免疫性疾病的治疗,对实验性自身免疫性葡萄膜炎(EAU)的治疗作用是目前研究的热点之一.目的研究雷帕霉素对EAU的治疗作用,并观察雷帕霉素对EAU各免疫细胞群炎性因子表达的影响.方法25只Lewis大鼠采用随机数字表法分为EAU组(20只)和正常对照组(5只).光感受器间维生素A类结合蛋白(IRBP) R16肽段与完全氟氏佐剂充分乳化后于Lewis大鼠后足皮下注射以建立EAU模型,EAU模型鼠再按分层随机的原则分为模型对照组和雷帕霉素组,每组10只大鼠.雷帕霉素组造模后即应用0.2 mg/(kg·d)雷帕霉素(0.4 ml)腹腔内连续注射7d,模型对照组及正常对照组大鼠采用等体积的生理盐水进行腹腔内注射.造模后第4天开始每日裂隙灯下观察大鼠EAU的症状,造模后第14天制备大鼠视网膜切片,以苏木精—伊红染色法进行组织病理学观察,参照Caspi的标准对EAU症状及组织病理学分级进行评分.应用免疫组织化学染色法检测各组大鼠视网膜中炎性因子干扰素-γ( IFN-γ)、白细胞介素-17( IL-17)的表达情况.结果造模后6d模型对照组大鼠EAU炎症评分逐渐升高,12 d达到高峰,然后逐渐下降.雷帕霉素组大鼠EAU炎症评分变化呈现相同的趋势,但各时间点EAU炎症评分均明显低于模型对照组,差异均有统计学意义(P＜0.01).视网膜组织病理学研究表明,模型对照组大鼠视网膜结构紊乱,大量炎性细胞浸润,组织病理学评分为3.30±0.48,而雷帕霉素组视网膜结构接近正常,组织学评分为0.90±0.45,差异有统计学意义(t=16.541,P＜0.01).雷帕霉素组IFN-γ、IL-17在大鼠视网膜中的表达量(A值)分别为21.16±4.23和49.86±6.59,明显低于模型对照组的62.14±7.32和124.85±6.33,差异均有统计学意义(q=33.334、q=56.923,P＜0.01).结论雷帕霉素通过抑制EAU视网膜中IFN-γ、IL-17等炎性因子的表达而对EAU发挥治疗作用,其机制可能是通过抑制Th1、Th17细胞群来实现的. 揹景雷帕黴素不僅具有抗菌作用,而且是一種良好的免疫抑製劑,可用于多種自身免疫性疾病的治療,對實驗性自身免疫性葡萄膜炎(EAU)的治療作用是目前研究的熱點之一.目的研究雷帕黴素對EAU的治療作用,併觀察雷帕黴素對EAU各免疫細胞群炎性因子錶達的影響.方法25隻Lewis大鼠採用隨機數字錶法分為EAU組(20隻)和正常對照組(5隻).光感受器間維生素A類結閤蛋白(IRBP) R16肽段與完全氟氏佐劑充分乳化後于Lewis大鼠後足皮下註射以建立EAU模型,EAU模型鼠再按分層隨機的原則分為模型對照組和雷帕黴素組,每組10隻大鼠.雷帕黴素組造模後即應用0.2 mg/(kg·d)雷帕黴素(0.4 ml)腹腔內連續註射7d,模型對照組及正常對照組大鼠採用等體積的生理鹽水進行腹腔內註射.造模後第4天開始每日裂隙燈下觀察大鼠EAU的癥狀,造模後第14天製備大鼠視網膜切片,以囌木精—伊紅染色法進行組織病理學觀察,參照Caspi的標準對EAU癥狀及組織病理學分級進行評分.應用免疫組織化學染色法檢測各組大鼠視網膜中炎性因子榦擾素-γ( IFN-γ)、白細胞介素-17( IL-17)的錶達情況.結果造模後6d模型對照組大鼠EAU炎癥評分逐漸升高,12 d達到高峰,然後逐漸下降.雷帕黴素組大鼠EAU炎癥評分變化呈現相同的趨勢,但各時間點EAU炎癥評分均明顯低于模型對照組,差異均有統計學意義(P＜0.01).視網膜組織病理學研究錶明,模型對照組大鼠視網膜結構紊亂,大量炎性細胞浸潤,組織病理學評分為3.30±0.48,而雷帕黴素組視網膜結構接近正常,組織學評分為0.90±0.45,差異有統計學意義(t=16.541,P＜0.01).雷帕黴素組IFN-γ、IL-17在大鼠視網膜中的錶達量(A值)分彆為21.16±4.23和49.86±6.59,明顯低于模型對照組的62.14±7.32和124.85±6.33,差異均有統計學意義(q=33.334、q=56.923,P＜0.01).結論雷帕黴素通過抑製EAU視網膜中IFN-γ、IL-17等炎性因子的錶達而對EAU髮揮治療作用,其機製可能是通過抑製Th1、Th17細胞群來實現的.
배경 뢰파매소불부구유항균작용,이차시일충량호적면역억제제,가용우다충자신면역성질병적치료,대실험성자신면역성포도막염(EAU)적치료작용시목전연구적열점지일.목적 연구뢰파매소대EAU적치료작용,병관찰뢰파매소대EAU각면역세포군염성인자표체적영향.방법25지Lewis대서채용수궤수자표법분위EAU조(20지)화정상대조조(5지).광감수기간유생소A류결합단백(IRBP) R16태단여완전불씨좌제충분유화후우Lewis대서후족피하주사이건립EAU모형,EAU모형서재안분층수궤적원칙분위모형대조조화뢰파매소조,매조10지대서.뢰파매소조조모후즉응용0.2 mg/(kg·d)뢰파매소(0.4 ml)복강내련속주사7d,모형대조조급정상대조조대서채용등체적적생리염수진행복강내주사.조모후제4천개시매일렬극등하관찰대서EAU적증상,조모후제14천제비대서시망막절편,이소목정—이홍염색법진행조직병이학관찰,삼조Caspi적표준대EAU증상급조직병이학분급진행평분.응용면역조직화학염색법검측각조대서시망막중염성인자간우소-γ( IFN-γ)、백세포개소-17( IL-17)적표체정황.결과조모후6d모형대조조대서EAU염증평분축점승고,12 d체도고봉,연후축점하강.뢰파매소조대서EAU염증평분변화정현상동적추세,단각시간점EAU염증평분균명현저우모형대조조,차이균유통계학의의(P＜0.01).시망막조직병이학연구표명,모형대조조대서시망막결구문란,대량염성세포침윤,조직병이학평분위3.30±0.48,이뢰파매소조시망막결구접근정상,조직학평분위0.90±0.45,차이유통계학의의(t=16.541,P＜0.01).뢰파매소조IFN-γ、IL-17재대서시망막중적표체량(A치)분별위21.16±4.23화49.86±6.59,명현저우모형대조조적62.14±7.32화124.85±6.33,차이균유통계학의의(q=33.334、q=56.923,P＜0.01).결론뢰파매소통과억제EAU시망막중IFN-γ、IL-17등염성인자적표체이대EAU발휘치료작용,기궤제가능시통과억제Th1、Th17세포군래실현적.
Background Studies determined that rapamycin has not only the antibiosis but also suppressing the auto-immunology.The treating effect of rapamycin on experimental autoimmune uveoretinitis (EAU) is still concerned.Objective This work was to investigate the therapeutic effect of rapamycin on EAU and study the effect on the expression of inflammatory cytokines which were secreted by T lymphocyte subgroup in EAU.Methods EAU was induced in 20 SPF male Lewis rats by subcutaneous injection of interphotoreceptor retinoid binding protein (IRBP) R16 peptide emulsified in adjuvant.The rats were randomized into model control group and rapamycin injection group and 10 rats for each group.The rapamycin of 0.2 mg/( kg · d) 0.4 ml was intraperitoneally injected for the consecutive 7 days immediately after modeling,and the equal volume of normal saline solution was used at the same fashion in the model control group and 5 normal matched rats( normal control group).The ocular manifestation of the rats were examined under the slit lamp regularly.The retinal sections of the rats were prepared in the 14 days after modeling for the histopathological examination with hematoxylin and eosin staining.The ocular signs of inflammation and histopathological severity were scored based on the criteria of Caspi.Expression of interferon-γ (IFN-γ) and interleukin-17 (IL-17) in rat retinas were detected by immunohistochemistry.This experiment followed the Regulation for the Administration of Affair Concerning Experimental Animals by Tianjin Municipal Government.Results The scores of ocular signs elevated gradually from 6 through 12 days after modeling and slowly declined after that in the model control group.A same tendency was seen in the rapamycin injection group.The significant differences were seen in the scores of ocular signs between the two groups from 6 to 14 days( P＜0.01 ).The disorder of retinal structure and infiltration of inflammatory cells were seen in the model control group,but the retina layers were normal in the rapamycin injection group.The pathological score was evidently declined in the rapamycin injection group ( 0.90 ± 0.45 ) in comparison with model control group( 3.30±0.48 ) ( t =16.541,P＜0.01 ).The expressions of the IFN-γ and IL-17 in retina located in the outer nuclear layer,inner nuclear layer,internal plexiform layer and retinal ganglion cell layer with the weakened levels(A values) in rapamycin injection group compared with model control group,showing a considerably difference between them ( IFN-γ:21.16±4.23 vs 62.14 ±7.32; IL-17:49.86±6.59 vs 124.85 ±6.33 )(q=33.334,q=56.923,P＜0.01 ).Conclusions Rapamycin down-regulates the expressions of IFN-γ and IL-17 in retina and further eliminates the inflammatory response in the rat with EAU by the suppression of Th1 and Th17 cells function in EAU.