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

对大鼠下肢进行同种异体移植（Fisher 344至刘易斯）和同种异体移植（刘易斯至刘易斯）。通过定量Western印迹分析和免疫组织化学进行DNA的时间分布。用TUNEL法检测移植物中细胞凋亡。术后即刻（第0天）和术后第1、3、7、12天（免疫组化法和Westerblot法各5例），同种异体移植物中DNASey的表达水平明显高于同种异体移植物（P< 0.001）。移植后第1天，移植物角质形成细胞、毛囊细胞和真皮下血管丛内皮细胞对DNA酶的免疫反应增强。我们还在DNA免疫组织化学染色阳性的细胞中检测到了凋亡性细胞死亡，这些结果提示在MHC相容性微小错配同种异体移植排斥反应过程中可能存在凋亡性细胞死亡。因此，调节细胞凋亡机制可能是改善MHC相容性微小错配同种异体移植物结局的重要治疗选择。

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.