蛋白激酶C-β抑制剂对大鼠移植肾的影响及磁共振成像在移植物中的变化

Effect of protein kinase C-βinhibitor on transplanted kidneys in rats and the changes of magnetic resonance imaging in the grafts

目的通过建立大鼠肾移植模型,探讨蛋白激酶C-β(PKC-β)抑制剂对大鼠移植肾的影响并观察磁共振成像(MRI)在移植物中的相应变化。方法建立大鼠肾移植模型,将其采用随机数字表法分为3组,同质移植对照组、异质移植对照组和PKC-β抑制剂治疗组,每组10对。术后24 h先行MRI检查,然后处死大鼠取标本。术后第1天取静脉血用酶联免疫吸附试验(ELISA)法检测血清中白细胞介素18(IL-18)、中性粒细胞明胶酶相关脂质运载蛋白(NGAL);术后第5天取静脉血检测各组血清肌酐(SCr)及尿素(BUN);肾组织做苏木精-伊红(HE)染色,过碘酸雪夫(PAS)染色;IHC检测肾损伤分子1(KIM-1)。组间比较采用单因素方差分析。结果 MRI显示术后24h大鼠移植肾在异质移植PKC-β抑制剂治疗组表观扩散系数(ADC)及T2加权成像分别为[(1.542±0.218)^(-3) mm^(2)/s、(58.7±4.9) ms],显著高于异质移植对照组[(1.437±0.198)^(-3) mm^(2)/s、(54.3±4.7) ms],而低于同质移植对照组[(1.692±0.235)×10^(-3) mm^(2)/s、(60.1±5.3) ms],差异有统计学意义(F=91.232、39.428,P值均<0.01);SCr、BUN、KIM-1、IL-18、NGAL在异质移植PKC-β抑制剂治疗组表达水平[(367.429±84.693) μmol/L、(49.629±14.506) mmol/L、(7.704±2.600)、(71.712±21.052) pg/ml、(222.805±131.799) pg/ml]分别高于同质移植对照组[(171.778±89.081) μmol/L、(26.807±12.468) mmol/L、(6.413±2.482)、(49.395±14.535) pg/ml、(157.618±30.439) pg/ml],但低于异质移植对照组[(529.667±68.661) μmol/L、(70.907±12.075) mmol/L、(9.869±3.758)、(111.838±18.663) pg/ml、(830.787±209.994) pg/ml],差异有统计学意义(F=70.782、45.398、51.396、58.113、78.455,P值均<0.01)。结论 PKC-β抑制剂可减轻肾移植后移植物的损伤,移植物通过无损伤MRI检测与上述结果呈对应关系。

Objective To investigate the effect of protein kinase C-β(PKC-β)inhibitor on transplanted kidneys in rats by establishing renal transplantation model and to observe the changes of magnetic resonance imaging(MRI)in the grafts.Methods The rats were randomly divided into three groups:isograft control group,allograft control group and allograft with PKC-βinhibitor treatment group(PKC group),n=10 each.MRI was performed 24 h after the operation,and then the animals were sacrificed for specimens.Enzyme linked immunosorbent assay(ELISA)was used to analyze serum interleukin-18(IL-18)and neutrophil gelatinase-associated lipocalin(NGAL)in venous blood at 1st day after the operation.Serum creatinine(SCr)and blood urea nitrogen(BUN)in each group were detected in venous blood samples on the 5th day after operation.Kidneys were stained with hematoxylin and eosin(HE)and periodic acid Schiff(PAS).Immunohistochemistry(IHC)was used to analyze kidney injury molecule-1(KIM-1).Results MRI showed that the apparent diffusion coefficient(ADC)and T2-weighted imaging of rat grafts at 24 h after kidney transplantation in the PKC group[(1.542±0.218)^(-3) mm^(2)/s,(58.7±4.9)ms]were significantly increased as compare with those in the allograft control group[(1.437±0.198)^(-3) mm^(2)/s,(54.3±4.7)ms)],but significantly decreased as compared with those in the isograft control group[(1.692±0.235)^(-3) mm^(2)/s,(60.1±5.3)ms,F=91.232,39.428,P<0.01].The levels of SCr,BUN,KIM-1,IL-18 and NGAL in the PKC group[(367.429±84.693)μmol/L,(49.629±14.506)mmol/L,(7.704±2.600),(71.712±21.052)pg/ml,(222.805±131.799)pg/ml]were significantly higher than those in the allograft control group[(171.778±89.081)μmol/L,(26.807±12.468)mmol/L,(6.413±2.482),(49.395±14.535)pg/ml,(157.618±30.439)pg/ml]and significantly lower than those in the isograft control group[(529.667±68.661)μmol/L,(70.907±12.075)mmol/L,(9.869±3.758),(111.838±18.663)pg/ml,(830.787±209.994)pg/ml,F=70.782,45.398,51.396,58.113,78.455,P<0.01].Conclusion PKC-βinhibitor can reduce the graft injury after kidney transplantation,and the results of non-invasive MRI scan for the grafts are correlated with above analysis.