大鼠心脏移植急性排斥反应过程中穿孔素及颗粒蛋白酶B mRNA的表达

Expression of Perforin and Granzyme B mRNA in Peripheral Blood Lymphocytes in Acute Cardiac Allograft Rejection in Rats

目的通过动态监测大鼠异体异位心脏移植外周血（PB）中穿孔素（pedorin，P）和颗粒酶B（granryme B，OB）的表达水平，判断其与急性排斥反应的相关性。方法实验组（n=16），供受体分别为Wistar和SD大鼠；对照组（n=17），供受体均为SD大鼠，两组术后均未接受抗排斥治疗。于术前及术后不同的时间采血，同时进行活组织病理检查，应用荧光定量RT-PCR（FQ-PCR）的方法，测定P及GBmRNA的表达水平，并与同期的病理检查结果相对照。结果实验组于移植后24hP及GBmRNA的表达上调，术后72h的表达明显增加（P〈0．001），术后3—5d达峰值，7—11d维持高水平，其变化早于病理改变；而对照组P及GBmRNA的表达及病理改变，在同期的观察过程中均没有明显的变化（P〉0．05）。结论同种异体大鼠心脏移植的急性排斥反应过程中，外周血淋巴细胞的穿孔素与颗粒酶BmRNA的表达水平术后呈现递增性，早于病理改变，可以作为急性排斥反应的监测指标。

Objective To observe the expression of pefforin and granzyme B mRNA in peripheral blood lymphocytes dynamically after heart transplantation in rats, and to assess the relationship between their expression level and acute cardiac allograft rejection. Methods The donor＇ s heart and up-right lung were transplanted into the recipient＇ s abdomen by the donor ascending aorta anastomized with recipient abdominal artery. The rats were divided into two groups after operation, the experimental group （n = 17）, the donors were Wistar and the recipients were SD rats. The control group （n = 18）, the transplantation was performed just between SD rats. FQ-PCR was used to quantify the pedorin and granzyme B mRNA level in peripheral blood lympho-cytes at varying times, meanwhile the graft was harvested to examine the pathological changes. Results At 24 hours after transplantation, the expression of perforin and granzyme B mRNA began to be up-regulated, reaching the peak at 3 to 5 days, and still kept a high level at 7-11 days. There was a significant difference in comparison with the level before operation and the expression level of control group （ P 〈 0.001 ）. Those changes occurred earlier than the pathological changes. No changes were observed in the control group. Conclusion The graft rejection is concomitant with the expression of perforin and granzyme B mRNA. The expression level of perforin and granzyme B mBNA is changed earlier than pathological changes. The measurement of perforin and granzyme B mRNA may be a good indicator of acute rejection.