定量检测血和尿中Cystatin C含量在肾移植患者肾功能评价中的应用

The application of quantitative detection of cystatin C in evaluation of renal function after renal transplantation

目的:建立定量检测半胱氨酸蛋白酶抑制剂C(cysC)水平的夹心ELISA方法,对肾移植患者的肾功进行监测,以期作为术后急性排斥反应(AR)及肾功能延迟恢复(DGF)发生的一项新的预警指标。方法:建立分泌抗人cys C单克隆抗体(mAb)的杂交瘤细胞株,制备腹水纯化mAbs并进行辣根过氧化物酶标记,建立检测cys C的夹心ELISA方法;应用此方法检测正常人及23例肾移植患者移植手术前后的血清(Scys C)及尿(Ucys C)标本中cys C的含量并分析其与血清肌酐(SCR)的相关性。结果:获得7株可稳定分泌抗cys CmAb的杂交瘤细胞。应用建立的ELISA方法检测cys C含量敏感度可达0.1μg/L。肾移植正常恢复(NC)组患者Scys C及Ucys C含量与SCR下降趋势一致。术后发生AR的患者Ucys C水平持续较高,直到术后21 d才出现下降。而发生DGF的患者同NC组患者相比术后4周内Scys C及Ucys C与SCR维持在较高水平,Ucys C的水平有逐渐升高的趋势,直到术后60 d才下降到与NC相近的水平。结论:建立了敏感稳定的检测cys C含量的ELISA方法,可用于检测Ucys C水平。而Scys C及Ucys C的持续升高可能提示肾移植患者术后AR或DGF的发生。

AIM： To establish ELISA method for quantitate the concentration of cystatin C（cys C） and to monitor the renal function of patients before and after renal transplantation.METHODS： Hybridomas secreting monoclonal antibodies（mAbs） against human cys C were produced and sandwich ELISA kit for quantitatively detecting cys C was established.Then the concentrations of serum cystatin C（Scys C） and urine cystatin C（Ucys C） from normal controls and 23 patients undergoing renal transplantation were detected and their relationship with serum creatinine（SCR） was analyzed.RESULTS： Seven hybridomas secreting anti-cys C mAbs were obtained.The sensitivity of the established ELISA kit reached 0.1 μg/L.The concentrations of Scys C and Ucys C of normal healthy controls were in accordance with other report.High correlations between Scys C or Ucys C and the level of SCR were observed（P0.01）.Rapid decline of Scys C and Ucys C concentrations was consistent with the decrease of SCR in the patients with normal course（NC） recovery after renal transplantation.However,Ucys C kept higher level within two weeks after the operation in patients with AR until the day 21.In patients with DGF,higher levels of Scys C,Ucys C and SCR were sustained within four weeks after renal transplantation.CONCLUSION： The sensitive ELISA kit for detection of cys C has been established.Importantly,there are the persistently high levels of Scys C and Ucys C in patients with AR or DGF,which can be used as a novel indicator for monitoring renal function after renal transplantation.