大鼠角膜移植术后植片内颗粒酶B和穿孔素的表达

Expression of PFP and GrB in cornea after rat penetrating keratoplasty

目的观察大鼠角膜移植术后植片内穿孔素(PFP)和颗粒酶B(GrB)的表达情况。方法以Wister大鼠为供体,SD大鼠为受体,常规行穿透角膜移植术,裂隙灯观察移植排斥情况。分别于移植术后第3、7、14、21d取角膜植片,HE染色及免疫组织化学方法检测植片内PFP和GrB的表达。结果角膜移植术后(7.63±0.74)d出现排斥反应。免疫组织化学结果可见,PFP、GrB阳性染色为棕色,表达于淋巴细胞、中性粒细胞及单核吞噬细胞的胞浆。当被分泌出后多分布于角膜上皮细胞间隙和细胞膜,正常角膜无表达。角膜移植术后第3d,大部分植片未见PFP和GrB的表达,少数植片见植片植床交界处上皮少量表达,第7d起表达明显,第14d达高峰,第21d稍有下降。PFP与GrB的表达具有相关性。结论PFP与GrB在大鼠角膜排斥反应中起着重要的作用。

Objective To observe the expression of pore-forming protein （PFP） and granzyme（GrB） in rat cornea after penetrating keratoplasty （PKP）. Methods Thirteen Wister rats and 26 SD rats were used as donors and as receptors respetively. Routine PKP was performed. Reject reaction was observed using slit lamp in 10 SD rats,recording rejective indicator （RI）. Four corneas were excised respectively at 3,7,14 and 21 days after surgery. HE staining and immunohistochemistry of PFP and GrB were performed. Results The reject reaction occurred at （7.63±0. 74） days after PKP. PFP and GrB were seen in the plasma of lymphocytic cell,neutrophilic granulocyte,mononuclear cell and cell membrane of cornea epithelium. There was no apparent expression of PFP and GrB in normal cornea. The expression was negligible at 3 days, elevated at 7 days, peaked at 14 days,and declined at 21 days. A correlation between PFP and GrB was found. Conclusion PFP and GrB play important roles in keratoplasty rejective reaction.