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目的探讨大鼠肾移植术后移植肾、外周血单核细胞(PBMCs)中穿孔素定量表达在急性排斥反应诊断中的意义。方法实验组由Wistar大鼠为供体,SD大鼠为受体。对照组供受体均为SD大鼠。术后第5天处死大鼠,切取移植肾组织行病理学检查和穿孔素mRNA定量检测,同时抽取外周血5ml定量检测穿孔素mRNA表达。比较2组的表达水平。结果实验组移植肾病理评分2~5分(3.43±0.98),对照组移植肾病理评分0~1分(0.71±0.49),2组比较差异有统计学意义(P<0.01)。对照组移植肾和外周血标本中穿孔素mRNA表达水平分别为(0.36±0.27)×106和(0.43±0.27)×106拷贝/μgRNA,实验组表达水平分别为(23.37±10.85)×106和(61.03±41.25)×106拷贝/μgRNA,差异有统计学意义(P<0.001)。结论移植肾组织和外周血单核细胞内穿孔素mRNA表达水平可作为急性排斥反应的诊断标志;外周血单核细胞可代替移植肾组织作为检测免疫活化基因mRNA表达水平的标本,实现急性排斥反应诊断的无创性要求。
Objective To investigate the significance of quantitative expression of perforin in renal allograft and peripheral blood mononuclear cells (PBMCs) after renal transplantation in the diagnosis of acute rejection. Methods Wistar rats were used as donors and SD rats as recipients. Control recipients were SD rats. The rats were sacrificed on the fifth day after operation. The pathological examination of the transplanted kidneys and perforin mRNA were performed. Quantitative detection of perforin mRNA expression by 5 ml of peripheral blood was also performed. The levels of expression in two groups were compared. Results In the experimental group, the grading of renal pathology was 2 to 5 points (3.43 ± 0.98) and the control group was 0 to 1 (0.71 ± 0.49) respectively. The difference between the two groups was statistically significant (P <0.01). The levels of perforin mRNA in the transplanted kidney and peripheral blood of the control group were (0.36 ± 0.27) × 106 and (0.43 ± 0.27) × 106 copies / μgRNA respectively, and the expression levels of the experimental group were (23.37 ± 10.85) × 106 and 61.03 ± 41.25) × 106 copies / μgRNA, the difference was statistically significant (P <0.001). Conclusion The expression of perforin mRNA in renal allograft and peripheral blood mononuclear cells can be used as a diagnostic marker for acute rejection. Peripheral blood mononuclear cells can be used as a sample for detecting the expression of immunoreactive gene mRNA in acute rejection Diagnostic non-invasive requirements.