SYBR Green Ⅰ荧光定量PCR检测肾移植患者IL-2和IL-10基因表达

SYBR GreenⅠfluoresence quantitative reverse transcriptase-polymerase chain reaction for measurement of IL-2 and IL-10 gene expressions in patients with renal transplantation

目的对比肾移植患者和常规手术患者免疫状态,验证以β-actin mRNA为模板参照定量PCR技术的有效性。方法采用SYBR GreenⅠ荧光定量PCR对29例肾移植患者和10例常规手术患者术前、术后3和7天外周血淋巴细胞IL-2和IL-10 mRNA表达进行检测。结果两组术前IL-2和IL-10 mRNA水平无差别;术后肾移植组IL-2mRNA明显降低,而对照组无明显变化;两组术后第3天IL-10 mRNA表达水平均明显升高,肾移植组高于对照组(P<0.01),术后第7天对照组降至正常,肾移植组亦下降,但水平仍高于术前。结论SYBR GreenⅠ荧光定量PCR技术对于监测肾移植患者免疫状况和研究免疫耐受意义重大,以β-actin mRNA为模板参照的定量PCR技术是测定细胞因子基因表达的简单有效方法。

Objective To compare the immunity status of the patients performed renal transplantation with the patients applied for routine operation, verify the effect of quantitative PCR which take β-actin mRNA as reference template. Methods By SYBR Green I fluoresence quantitative PCR technique, we measured IL-2 and IL-10 mRNA expressions of peripheral blood lymphocytes from 29 patients with renal transplantation （RT） and 10 patients with routine operation （control group） , at the times of before operation, 3 days and 7 day3 after operation. Results IL-2 and IL-10 mRNA in two groups had no difference before operation. After operation, IL-2 mRNA deceased apparently in RT group, while no significant change happened in control group. 3 days after operation, IL-10 mRNA impression both increased significantly, it was higher in KT group than that of control group. 7 days after operation, it decreased to normal level in control group, also it decreased in RT group, but still higher than that before operation. Conclusions SYBR Green I fluoresence quantitative PCR technique is beneficial for measuring immunity status and studying immunity tolerance in patients with renal transplantation; The quantitative PCR technique taking β-actin mRNA as reference template is a simple and effective method for measuring the expressions of cytokines gene.