外周血穿孔素和颗粒酶B mRNA检测对胰腺移植急性排斥反应早期诊断的价值

目的 研究胰腺移植术后外周血穿孔素和颗粒酶B的基因表达对急性排斥反应早期诊断的价值.方法 实验动物为杂种长白猪,分为3组(n=8):对照组,移植组.免疫抑制剂治疗组.移植手术为同种异体门静脉回流、肠内引流全胰十二指肠移植.免疫抑制剂治疗组在行同种异体胰十二指肠移植的同时应用环孢A 骁悉 甲强龙三联免疫抑制治疗.分别于术前1 d和术后1、3、5、7 d采集受者锁骨下静脉血,抽提总RNA,RT.PCR方法检测静脉血淋巴细胞中穿孔素和颗粒酶B的mRNA表达,检测受者血糖、胰岛素、胰高血糖素水平.在术后1、3、5、7d,开腹取胰腺组织进行常规病理学检查,与血液中穿孔素和颗粒酶B的mRNA表达水平进行比较分析.结果 ①对照组、移植组、免疫抑制剂组之间受体的血糖、胰岛素、胰高血糖素水平变化差异无统计学意义.②与移植组比较,免疫抑制剂组静脉血淋巴细胞中穿孔索和颗粒酶B的mRNA表达水平下降(P<0.05).③胰腺移植术后静脉血穿孔素、颗粒酶B mRNA表达变化比急性排斥反应病理学改变早2～d.结论 监测外周血淋巴细胞中穿孔素和颗粒酶B mRNA表达的变化,可以对胰腺急性排斥反应做出早期诊断.

Perforin and Granzyme B as An Early Indicator for Acute Rejection of Pancreatic Transplantation

Objective To assess the value of perforin mRNA and granzyme B mRNA in lymphocytes as an early diagnostic indicator for acute rejection of pancreatic transplantation.Methods Twenty-four hybrid landraces were divided into three groups(control group,transplantation group,transplantation plus immunosuppressant group) randomly.The landraces in the transplantation group were treated with pancreatoduodenal allotranplantation.The landraces in the transplantation plus immunosuppressant group underwent the same surgery and were intervened simultaneously with CsA,MMF,and Metrisone.Peripheral venous bloods were sampled via subclavic veins one day before the operations and on the 1st,3rd,5th and 7th day after the operations.The RNA was extracted from the blood samples.The mRNA of perforin and granzyme B expressed in the lymphocytes were detected by RT-PCR.The serum glucose,insulin and glucagons were also measured with routine methods.The pancreatic graft tissues were collected for pathologic examinations at the 1st,3rd,5th and 7th day after the operations.Results No differences in serum glucose,insulin and glucagons had been detected among the three groups(P>0.05).The landraces in the transplantation plus immunosuppressant group had significantly lower expression of the mRNA of perforin and granzyme B in the lymphocytes and the local transplanted pancreas than those in the transplantation group(P<0.05).The change of mRNA expression of perforin and granzyme B in the lymphocytes occurred 2-4 days earlier than the pathological changes when the acute rejection started.Conclusion Monitoring the change of mRNA expression of perforin and granzyme B in the lymphocytes from the peripheral blood can help the early diagnosis of acute rejection of pancreatic transplantation.