急性排斥期大鼠角膜移植物基质细胞凋亡的研究

Study on apoptosis of keratocytes in the rat corneal grafts during the acute rejection

目的 研究急性排斥期角膜移植物基质细胞凋亡发生的规律 ,分析基质细胞凋亡与急性免疫排斥反应的关系。方法 建立大鼠穿透性角膜移植动物模型。实验分组 :A组 ,同种异体角膜移植组 ,Wistar→SD ;B组 ,同基因角膜移植组 ,Wistar→Wistar ;2移植组受者均为 10只。C组 ,正常对照非移植组 ,健康Wistar大鼠 10只。急性排斥期取材 ,应用细胞凋亡原位末端标记 (TUNEL法 )检测角膜基质细胞凋亡情况。对TUNEL染色结果采用全自动图像分析系统处理 ,计算阳性单位PU值。结果 与正常角膜及同基因角膜移植片相比 ,急性排斥期同种异体角膜植片基质细胞凋亡明显增多。各实验组阳性单位PU平均值分别为 :2 8 6 8± 3 6 4 ,10 80± 3 2 0 ,均明显高于对照组 2 72± 2 0 8,差异有显著性 (P <0 0 1)。两实验组相比 ,PU值差异有显著性 (P <0 0 1)。结论 细胞凋亡可能是导致角膜移植排斥反应发生的主要因素之一。

Objective To study apoptosis of keratocytes in corneal grafts during acute rejection period.Methods Corneal transplantation was performed orthotopically.Two groups were allograft(Wistar to SD rats)and autograft(Wistar to Wistar rats)as experimental groups,ten recipients for each experimental group;and the normal rats(ten Wistar rats)as control. During the acute immune rejection, the corneal samples were removed and fixed. The apoptosis in keratocyte was studied by TUNEL assay. An automatic image analyzer was used to analyze the staining results of TUNEL assay. Results The TUNEL positive cells of the allograft group was significantly higher compared with the control group and autograft group,respectively. The positive PU of 2 experimental groups was 28 68±3 64 and 10 80±3 20,which was higher than control's 2 72±2 08,the difference was significant( P <0 01).Compared with each other,the difference of positive PU of 2 experimental groups was significant( P <0 01).Conclusion Apoptosis plays an important role in corneal allograft rejection.