移植肾功能延迟恢复患者尿脱落细胞中穿孔素和粒酶BmRNA定量检测的意义

Analysis of granzyme B mRNA and perforin mRNA levels in urine for renal transplantation patients with delayed graft fanction

目的探讨尿脱落细胞中穿孔素mRNA、粒酶BmRNA检测在移植肾功能延迟恢复（DGF）诊断中的应用价值。方法同种异体肾移植DGF患者24例。男15例，女9例。平均年龄37岁。移植肾活检病理分析病因为输尿管梗阻2例、静脉血栓1例、急性环孢素（CsA）中毒3例、急性肾小管坏死（ATN）7例、ATN合并临界改变2例、ATN合并急性排斥反应（AR）3例、AR6例。移植术后1～14d留取24例患者73份晨尿标本，应用竞争R^PCR方法对患者尿脱落细胞中穿孔素和粒酶BmRNA进行定量检测，应用SPSSl3．0软件进行数据分析，mRNA水平用自然对数值表示。结果24例患者尿脱落细胞中穿孔素和粒酶BmRNA水平在急性CsA中毒、ATN和其他组分别为（-0．76土0．35）、（-0．89±0．30）、（-0．90±0．15）fg／μg和（-0．53±0．22）、（-0．41±0．17）、（-0．73±0．23）fg／μg，组间两两比较差异均无统计学意义（P〉0．05）；ATN合并AR组与单纯AR组分别为（1．20士0．39）、（11．13±0．33）fg／μg和（1．07±0．30）、（1．01±0．19）fg／μg，2组比较差异无统计学意义（P〉0．05）；ATN合并AR、单纯AR组与急性CsA中毒、ATN和其他组比较差异有统计学意义（P〈0．001）；2例ATN合并临界改变患者尿中穿孔素和粒酶BmRNA平均值分别为1．22、0．97fg／μg，与ATN合并AR、单纯AR组相近。结论DGF患者尿脱落细胞中穿孔素和粒酶BmRNA水平检测有助于DGF的病因诊断，可作为临界改变是否需要进-步干预性治疗的指标之-。

Objective To explore the clinical value of the level of granzyme B and perforin mRNA in urine for the diagnosis of renal＇ transplantation patients with delayed graft function （DGF）. Methods Twenty-four cases of renal transplantation patients with DGF were included in this study. Seventy-three u- rine specimens were obtained from these patients who received graft biopsies. Among the 24 cases, ureteral obstruction occurred in 2 cases, vascular thrombosis in 1 case, acute CsA intoxication in 3 cases, acute tubu- lar necrosis （ATN） in 7 cases, ATN complicating borderline change in 2 cases, ATN complicating acute re- jection （AR） in 3 cases, AR in 6 cases. Total RNA was isolated from the urinary ceils. Messenger RNA （mRNA） encoding the cytotoxic proteins perforin and granzyme B gene were measured with the quantitative polymerase-chain-reaction assay （RT-PCR）. SPSS13. 0 software was used for data analysis. Levels of mRNA were log-transformed before analysis. Results The levels of perforin and granzyme B mRNA in u- rine among the ureteral obstruction group, vascular thrombosis group, acute CsA intoxication group and ATN group were very low. There was no significant difference among these groups （P〉0.05）. However, among the ATN complicating borderline change group 1.22, 0. 97 fg/μg, ATN complicating AR group （1.20±0. 39）, （1.07±0. 30）fg/μg, and AR group（11. 13±0.33）, （1.01±0. 19）fg/μg, the levels were increased significantly（P〉0. 001）. Conclusion Measurement of mRNA encoding the cytotoxie proteins perforin and granzyme B gene in urinary cells in renal transplantation patients with DGF could be helpful to etiological diagnosis of DGF, and might be used as an index for the appropriate management of the borderline change.