中华眼科杂志
中華眼科雜誌
중화안과잡지
Chinese Journal of Ophthalmology
2013年
10期
921-926
,共6页
马明%吴京%黄晓环%熊柯%陈林江
馬明%吳京%黃曉環%熊柯%陳林江
마명%오경%황효배%웅가%진림강
角膜移植%抗原,CD80%免疫耐受%移植物存活%小鼠
角膜移植%抗原,CD80%免疫耐受%移植物存活%小鼠
각막이식%항원,CD80%면역내수%이식물존활%소서
Corneal transplantation%Antigens,CD80%Immune tolerance%Graft survival%Mice
目的 探讨共刺激分子B7-H3在小鼠角膜中的表达情况以及在角膜移植免疫赦免中的作用.方法 实验研究.以C57BL/6小鼠为供体、BALB/c小鼠为受体建立角膜移植实验模型,采用简单随机抽样方法,取8只未行角膜移植的BALB/c小鼠作为正常对照组,另取8只BALB/c小鼠作为自体角膜移植组,最后30只BALB/c小鼠进行同种异体角膜移植；按Sonoda法观察小鼠角膜移植后的存活率,8周内植片混浊评分≥2分判定为排斥组,8周时未达到2分的认为是存活组；各组各取3只眼球,HE染色后行组织病理学观察并行B7-H3分子的免疫组织化学检测；各组各取5只眼角膜,行荧光定量PCR检测B7-H3分子mRNA的表达情况.各组角膜组织中的B7-H3 mRNA表达差异倍数比较采用设计单因素的方差分析,组间的多重比较采用LSD-t检验.结果 同种异体角膜移植组并未完全排斥,30只小鼠中有9只存活,21只排斥,移植存活率为30％；自体角膜移植组移植存活率为100％；免疫组织化学表明B7-H3分子在正常对照组和自体角膜移植组的角膜上皮、内皮细胞和虹膜睫状体中均有表达,在存活组中的表达明显增加,而在排斥组中的表达明显减少；qPCR检测在正常对照组(4.30±0.023)和自体角膜移植组(4.33±0.031)中均可以检测出B7-H3分子mRNA的表达；同种异体角膜移植排斥组B7-H3分子的mRNA表达程度(3.89±0.037)明显下降；但在同种异体角膜移植存活组中B7-H3分子的mRNA表达(5.04±0.058)明显增加；将各组B7-H3 mRNA的表达差异进行统计学分析,先进行方差齐性检验(F =429.546),再采用LSD法进行两两比较,除了正常对照组与自体角膜移植组之间差异无统计学意义(P =0.387)之外,正常对照组与存活组及排斥组比较,差异均有统计学意义(P =0.001,0.003)结论 B7-H3分子在促进角膜移植的免疫赦免中发挥一定的作用,可能是维持移植角膜免疫耐受状态的重要因子之一.
目的 探討共刺激分子B7-H3在小鼠角膜中的錶達情況以及在角膜移植免疫赦免中的作用.方法 實驗研究.以C57BL/6小鼠為供體、BALB/c小鼠為受體建立角膜移植實驗模型,採用簡單隨機抽樣方法,取8隻未行角膜移植的BALB/c小鼠作為正常對照組,另取8隻BALB/c小鼠作為自體角膜移植組,最後30隻BALB/c小鼠進行同種異體角膜移植；按Sonoda法觀察小鼠角膜移植後的存活率,8週內植片混濁評分≥2分判定為排斥組,8週時未達到2分的認為是存活組；各組各取3隻眼毬,HE染色後行組織病理學觀察併行B7-H3分子的免疫組織化學檢測；各組各取5隻眼角膜,行熒光定量PCR檢測B7-H3分子mRNA的錶達情況.各組角膜組織中的B7-H3 mRNA錶達差異倍數比較採用設計單因素的方差分析,組間的多重比較採用LSD-t檢驗.結果 同種異體角膜移植組併未完全排斥,30隻小鼠中有9隻存活,21隻排斥,移植存活率為30％；自體角膜移植組移植存活率為100％；免疫組織化學錶明B7-H3分子在正常對照組和自體角膜移植組的角膜上皮、內皮細胞和虹膜睫狀體中均有錶達,在存活組中的錶達明顯增加,而在排斥組中的錶達明顯減少；qPCR檢測在正常對照組(4.30±0.023)和自體角膜移植組(4.33±0.031)中均可以檢測齣B7-H3分子mRNA的錶達；同種異體角膜移植排斥組B7-H3分子的mRNA錶達程度(3.89±0.037)明顯下降；但在同種異體角膜移植存活組中B7-H3分子的mRNA錶達(5.04±0.058)明顯增加；將各組B7-H3 mRNA的錶達差異進行統計學分析,先進行方差齊性檢驗(F =429.546),再採用LSD法進行兩兩比較,除瞭正常對照組與自體角膜移植組之間差異無統計學意義(P =0.387)之外,正常對照組與存活組及排斥組比較,差異均有統計學意義(P =0.001,0.003)結論 B7-H3分子在促進角膜移植的免疫赦免中髮揮一定的作用,可能是維持移植角膜免疫耐受狀態的重要因子之一.
목적 탐토공자격분자B7-H3재소서각막중적표체정황이급재각막이식면역사면중적작용.방법 실험연구.이C57BL/6소서위공체、BALB/c소서위수체건립각막이식실험모형,채용간단수궤추양방법,취8지미행각막이식적BALB/c소서작위정상대조조,령취8지BALB/c소서작위자체각막이식조,최후30지BALB/c소서진행동충이체각막이식；안Sonoda법관찰소서각막이식후적존활솔,8주내식편혼탁평분≥2분판정위배척조,8주시미체도2분적인위시존활조；각조각취3지안구,HE염색후행조직병이학관찰병행B7-H3분자적면역조직화학검측；각조각취5지안각막,행형광정량PCR검측B7-H3분자mRNA적표체정황.각조각막조직중적B7-H3 mRNA표체차이배수비교채용설계단인소적방차분석,조간적다중비교채용LSD-t검험.결과 동충이체각막이식조병미완전배척,30지소서중유9지존활,21지배척,이식존활솔위30％；자체각막이식조이식존활솔위100％；면역조직화학표명B7-H3분자재정상대조조화자체각막이식조적각막상피、내피세포화홍막첩상체중균유표체,재존활조중적표체명현증가,이재배척조중적표체명현감소；qPCR검측재정상대조조(4.30±0.023)화자체각막이식조(4.33±0.031)중균가이검측출B7-H3분자mRNA적표체；동충이체각막이식배척조B7-H3분자적mRNA표체정도(3.89±0.037)명현하강；단재동충이체각막이식존활조중B7-H3분자적mRNA표체(5.04±0.058)명현증가；장각조B7-H3 mRNA적표체차이진행통계학분석,선진행방차제성검험(F =429.546),재채용LSD법진행량량비교,제료정상대조조여자체각막이식조지간차이무통계학의의(P =0.387)지외,정상대조조여존활조급배척조비교,차이균유통계학의의(P =0.001,0.003)결론 B7-H3분자재촉진각막이식적면역사면중발휘일정적작용,가능시유지이식각막면역내수상태적중요인자지일.
Objective To investigate the role of B7-H3 in the immune reaction of corneal transplantation in mice Methods Experimental study.Thirty Corneas of C57BL/6 mice were orthotopically transplanted into the eyes of BALB/c mice,and graft survival was assessed on the basis of Sonoda's standard.When the RI grade was ≥2,rejection was acknowledged and brought into the rejected group(R),and the others into the accepted group(A) ; 8 BALB/c Corneas into their own eyes belonged to isografts(Ⅰ) ;In addition,8 normal BALB/c mice were the control group(C).At last,three eyes in each group(C,I,A,R groups) were used for HE staining and IHC of B7-H3,and there were five eyes in each group for qPCR to detect B7-H3 mRNA expression.Repeated-measures analysis of variance (factorial analysis) followed by LSD test were used for post hoc analysis for expression differences of B7-H3 mRNA between groups.Results There were 9 accepted grafts and 21 rejected grafts in 30 mice,and transplantation survival rate was 30％ in the allograft group,while all grafts were transparent,and transplantation survival rate was 100％ in the isografts.IHC results showed that B7-H3 was expressed on the corneal epithelium,endothelium and irisciliary body of both normal corneas and Isografts; B7-H3 expression increased in the accepted group and decreased in the rejected group.The results of qPCR conformed to the IHC ; Repeated-measures analysis of variance (factorial analysis) followed by LSD test were used for post hoc analysis for differences between groups(F =429.546).there was a low B7-H3 mRNA expression in the R group(3.89 ±0.037) and high expression in the A group (5.04 ± 0.058) ; and C (4.30 ± 0.023),I (4.33 ± 0.031) groups had no significant difference (P =0.387) ; But there was a significant difference between group R and C (P =0.003) or group A and C (P =0.001).Conclusion All above show that B7-H3 may play an important role in the maintenance of ocular immune privilege.
