SYBR GreenⅠ荧光定量PCR检测肾移植患者CsA效用

SYBR GreenⅠ fluoresence quantitative reverse transcriptase polymerase chain reaction for measurement of cyclosporine A effect in patients with renal transplantation

  • 摘要: 目的: 检测肾移植患者外周血淋巴细胞(PBL)有关细胞因子基因表达,判定环孢素A(CsA)效用,进一步验证CsA服用后2h浓度(C2)测定的必要性。方法: 采用SYBR GreenⅠ荧光定量PCR对20例肾移植患者术后第7天PBL的IL-2和IFN-γmRNA表达进行检测,判定CsA的免疫抑制效用,对比CsA空腹浓度(C0)和C2与细胞因子抑制关系。结果: C2对应IL-2和IFN-γmRNA/β-actin mRNA相对模板数分别为0.038±0.006和0.018±0.004,C2对IL-2和IFN-γmRNA表达抑制率分别为(72.42±3.95)%和(69.52±6.97)%;C2与IL-2和IFN-γmRNA表达的抑制率有良好的线性关系(P<0.01);C0与IL-2和IFN-γmRNA表达的抑制率无良好的线性关系(P(0.05)。结论: SYBR GreenⅠ荧光定量PCR技术检测肾移植患者术后PBL的IL-2和IFN-γmRNA的表达,是判定CsA效用和研究免疫耐受的有效方法;C2仍为判定CsA生物利用度的重要指标。

     

    Abstract: Objective: To measure expressions of cytokines genes in peripheral blood lymphocyte of patients after renal transplantation, evaluate cyclosporine A effect, further verify the necessity of testing cyclosporine A concentration 2. Methods: By SYBR GreenⅠ PCR technique, we measured IL-2 and IFN-γ mRNA expressions of PBL from 20 cases of patients 7 days after renal transplantation, assessed immunosuppressive effect of cyclosporine A, contrasted the relation of C0 and C2 with cytokines inhibiting. Results: Corresponding to C2, IL-2 and IFN-γ mRNA/β-actin mRNA relative templates were 0.038± 0.006 和0.018±0.004 respectively,IL-2 and IFN-γ inhibiting rates were (72.42±3.95)%和(69.52±6.97)%. linear correlation of C2 with inhibiting rate of IL-2、IFN-γ mRNA expression was fine(P<0.01), but the one of C0 with them was bad (P>0.05). Conclusions: To measure IL-2 and IFN-γ mRNA expressions in PBLof patients after renal transplantation with SYBR GreenⅠ PCR technique is an effective method to evaluate cyclosporine A effect and immunosuppressive study, C2 is still an important index to judge biological availability of cyclosporine A.

     

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