外周血淋巴细胞穿孔素和颗粒酶B mRNA检测对肾移植急性排斥反应的早期诊断价值

The early diagnostic value of perforin and granzyme B mRNAs of peripheral blood lymphocytes in detection of renal allograft acute rejection

探讨外周血淋巴细胞(peripheral blood lymphocyte,PBL)穿孔素和颗粒酶B mRNA检测对肾移植急性排斥反应(AR)的早期诊断价值。采用RT-PCR方法动态检测67例肾移植AR患者外周血淋巴细胞穿孔素和颗粒酶B mRNA的表达水平,同时检测所有患者肌酐(Cr)表达水平情况,比较各时间点穿孔素、颗粒酶B mRNA及Cr表达水平,并探讨穿孔素、颗粒酶B与AR的关系。移植后1d与移植前1d比较,PBL穿孔素、颗粒酶B mRNA表达水平比较无明显差异性(P>0.05),Cr表达水平明显下降(P<0.05);AR前3d较之前各时间点比较,PBL穿孔素、颗粒酶B mRNA表达水平明显上升(P<0.05),而Cr表达水平明显下降(P<0.05);AR后1d时PBL穿孔素、颗粒酶B mRNA表达水平达到最高峰,随后逐渐下降(P<0.05),而AR后5d时Cr表达水平达到最高峰,随后逐渐下降(P<0.05);PBL穿孔素、颗粒酶B mRNA表达水平升高时间比Cr早4d,AR患者经MP/OKT3冲击治疗后,PBL穿孔素和颗粒酶B mRNA表达水平逐渐降低至原有基础水平。PBL穿孔素和颗粒酶B mRNA在AR早期诊断和抗排斥反应疗效评估等方面具有一定的临床价值。

To investigate the early diagnostic value of perforin and granzyme B mRNA of peripheral blood lymphocytes（PBL） in detection of renal allograft acute rejection（AR）, perforin and granzyme B mRNAs expression of PBL in 67 patients with renal allograft AR were dynamically measured by quantitative reverse transcription PCR （RT-PCR）. At the same time, the creati- nine（Cr） was examined in all patients. The levels of mRNA of perforin and granzyme B as well of Cr value at different time points were compared, and the relationship of perforin, granzyme B and AR was analyzed. When the levels of the 3 indeces on day 1 after transplantation was compared with those on day 1 after transplantation, mRNA of perforin and granzyme B remained unchanged （P〉0.05） whereas the value of Cr decreased significantly（P〈0.05）. Three days prior to AR took place, mRNA levels of perforin and granzyme B were increased significantly （P〈0.05） compared with those of all previous time points, and while the level of Cr was decreased significantly （P〈0.05）. The levels of mRNA of PBL perforin and granzyme B reached the peak ld after AR, then decreased gradually （P〈0.05）, but the level of Cr reached the peak 5 days after AR, then decreased gradually （P〈0.05）. Therefore the increase of perforin and granzyme B mRNA appeared 4 days earlier than Cr. The levels of perforin and granzyme B mRNA decreased to the original basic levels by MP/OKT3 pulse therapy. These results showed that the levels of perforin and granzyme B mRNA of PBL may be a noninvasive, sensitive indices for early diagnosis of AR and for assessing clinic effect of anti-rejection therapy, and It may be worthy of clinical application.