穿孔素和颗粒酶B蛋白表达对胰腺移植急性排斥反应早期诊断的研究

目的 研究移植胰腺局部穿孔素和颗粒酶B的蛋白表达对急性排斥反应的早期诊断和鉴别诊断的意义.方法 将40只四川当地杂种长白猪随机分为3组,即对照组(n=8)、移植组(n=16)、移植 免疫抑制剂治疗组(n=16).移植手术为同种异体经门静脉回流、肠内引流全胰十二指肠移植.免疫抑制剂治疗组在进行同种异体胰十二指肠移植术同时应用环孢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检测受者的锁骨下静脉血胰岛素、胰高血糖素、血糖水平.术后1、3、5、7 d,开腹取胰腺组织进行常规病理学检查和免疫组化染色检测穿孔素和颗粒酶B的蛋白表达.结果 ①对照组、移植组以及移植 免疫抑制剂组之间受者的血糖、胰岛素、胰高血糖素水平变化有统计学意义(P＜0.05).②胰腺移植 免疫抑制剂组与单纯胰腺移植组相比胰腺组织中穿孔素和颗粒酶B的蛋白表达及穿孔素和颗粒酶B的阳性淋巴细胞计数的差异有统计学意义(P＜0.05).③免疫组化染色检测移植后胰腺组织中穿孔素和颗粒酶B蛋白表达,比常规病理学检查诊断排斥反应早2～4 d.结论 移植胰腺组织穿孔素和颗粒酶B局部蛋白表达变化早于常规病理学改变,结合常规病理学检查可以对胰腺急性排斥反应做出早期诊断和鉴别诊断.

The Experimental Research on the Perforin and Granzyme B Expressions to the Early Diagnostic Value of Acute Rejection Response after Pancreatic Transplantation

OBJECTIVE: To study the early diagnosis value of acute rejection response to perforin and granzyme B protein expressions around transplantation pancreas tissue after pancreatic transplantation. METHODS: Twenty-four hybrid landraces were randomly divided into three groups [control group ( I group), transplantation group ( II group), transplantation + immunosuppressive group (III group)]. The transplantation group was treated by pancreatoduodenal allotransplantation, and simultaneously the group of transplantation + immunosuppressants was intervened by cyclosporin A (CsA), mycophenolate mofetil (MMF) and metrisone. The peripheral venous blood was sampled via subclavian vein one day before operation and on the 1st, 3rd, 5th or 7th day after operation respectively, and then the glucose, insulin and glucagons in serum were measured. The grafted pancreatic tissues were collected for pathologic and immunohistochemical examinations at 1st, 3rd, 5th and 7th day after operation. RESULTS: (1) The levels of blood sugar, insulin and glucagons had no significant difference among groups of control, transplantation and transplantation + immunosuppressant group (P>0.05). (2) By comparing with two experimented groups, the expressions of perforin and granzyme B protein in transplantation pancreas tissue had a significant difference between transplantation group and transplantation + immunosuppressive group (P < 0.05). (3) The time of perforin and granzyme B protein expressions in transplantation pancreas tissue after transplantation was 2-4 days earlier than that of pathological changes in acute rejection. CONCLUSION: The expression alterations of perforin and granzyme B protein are earlier appearance than the pathological changes of acute rejection response in transplantation pancreas tissue, and by combining with routine pathological examination, the acute rejection response can be diagnosed on early stage.