RESEARCH ON APOPTOTIC CELL DEATH IN THE RAT HINDLIMB ALLOTRANSPLANTATION MODEL OF ACUTE REJECTION

进行大鼠同种异体肢体移植(Fisher 344对Lewis)和同种异体移植(Lewis对Lewis)。用定量Western印迹分析和免疫组织化学方法检测DNASEY的时间分布。此外，我们还进行了原位末端标记法(TUNEL)检测移植物中的细胞凋亡。在移植后即刻(0.001天)、术后1、3、7、12天(Wester印迹定量分析N=5，免疫组织化学N=5)进行分析，与等密度梯度法相比，DNASE Y的表达水平在移植后7天均显著增强(P<0.05)。同种异体皮肤角质形成细胞、毛囊细胞和真皮下血管丛内皮细胞对DNASE的免疫反应性在移植后第1天增强。同时，在DNASE免疫组织化学阳性的同类细胞中也检测到了凋亡细胞的死亡。这些结果提示，凋亡细胞死亡可能参与了MHC相合的小失配同种异体排斥反应。因此，调节细胞凋亡机制可能是改善MHC相合的小失配同种异体移植物预后的重要治疗选择。

ALLOTRANSPLANTATION (FISHER 344 TO LEWIS) AND ISOTRANSPLANTATION (LEWIS TO LEWIS) OF THE RAT LOWER EXTREMITIES WERE PERFORMED. TEMPORAL PROFILES OF DNASEY WERE CONDUCTED BY QUANTITATIVE WESTERN BLOT ANALYSIS AND IMMUNOHISTOCHEMISTRY. ALSO, WE PERFORMED TUNEL TO DETECT APOPTOTIC CELL DEATH IN THE GRAFTS. ANALYSES WERE PERFORMED IMMEDIATELY AFTER REPER-FUSION (DAY 0), AND ON POSTOPERATIVE DAYS (PODS) 1, 3, 7,12 (N = 5 FOR QUANTITATIVE WESTER BLOT ANALYSIS AND N = 5 FOR IMMUNOHISTOCHEMISTRY).THE EXPRESSION LEVELS OF DNASEY WERE UNIFORMLY AND SIGNIFICANTLY AUGMENTED UNTIL POD 7 IN THE ALLOGRAFTS, COMPARED WITH ISOGRAFTS (P< 0.001). IMMUNOREACTIVITIES TO DNASEY IN KERATINOCYTES, HAIR FOLLICULAR CELLS AND ENDTHELIAL CELLS OF SUBDERMAL VASCULAR PLEXUS WERE ENHANCED IN THE ALLOGRAFTS ON POD 1. ALSO, WE DETECTED APOPTOTIC CELL DEATH IN THE SAME TYPES OF CELLS POSITIVE TO DNASEY IMMUNOHISTOCHEMISTRY.THESE RESULTS SUGGEST THE POSSIBLE INVOLVEMENT OF APOPTOTIC CELL DEATH IN THE PROCESS OF MHC-COMPATIBLE MINOR-MISMATCH ALLOGRAFT REJECTION. HENCE, MODULATION OF MECHANISMS OF APOPTOSIS MIGHT BE AN IMPORTANT THERAPEUTIC OPTION FOR IMPROVING THE OUTCOME OF MHC-COMPATIBLE MINOR-MISMATCH ALLOGRAFTS.