蛋白激酶C-β抑制剂减轻大鼠肾移植物的炎性浸润及与磁共振成像的对应关系

Protein kinase C-βinhibitor attenuates infiltration in rat kidney grafts and the correspondence with magnetic resonance imaging

目的通过制作大鼠肾移植模型,探讨蛋白激酶C-β(PKC-β)抑制剂对大鼠移植炎性反应的影响并观察与磁共振成像(MRI)的对应关系。方法将大鼠肾移植模型随机分为3组:同质移植组、异质移植组和异质移植PKC-β抑制剂组,每组10对。术后24 h行MRI检查,然后处死大鼠,取血标本用反转录-聚合酶链反应(RT-PCR)法检测低氧诱导因子-1(HIF-1)和血红素氧合酶-1(HO-1)的mRNA表达量,取肾脏标本用免疫组织化学检测肾组织中白细胞介素(IL)-4、IL-8、单核细胞趋化蛋白-1(MCP-1)和单核细胞迁移抑制因子(MIF)的表达。组间比较采用单因素方差分析。结果MRI显示术后24 h大鼠移植肾在异质移植PKC-β抑制剂组冠状位扫描记录动脉自旋标记(ASL)及T1成像分别为[(294.19±42.13)ml/(min×100 g)、(1951.1±82.9)ms],显著高于异质移植组[(253.24±25.18)ml/(min×100 g)、(1842.1±79.6)ms],而低于同质移植组[(331.65±49.44)ml/(min×100 g)、(2047.6±89.3)ms],差异有统计学意义(F=42.323、95.445,P值均<0.01);血清中的HIF-1和HO-1在异质移植PKC-β抑制剂组水平[(1245.17±305.96)、(249.72±51.01)]分别高于异质移植组[(947.34±112.65)、(111.71±41.02)]和同质移植组[(1157.74±224.63)、(147.52±42.22)],差异有统计学意义(F=61.252、37.832,P值均<0.01);而肾组织中的IL-4、IL-8、MCP-1、MIF在异质移植PKC-β抑制剂组表达水平[(45.3±4.3)、(47.2±4.5),(37.7±3.6)、(31.2±3.0)]分别高于同质移植组[(19.4±2.6)、(19.6±2.5)、(17.4±2.1)、(16.7±2.5)],但低于异质移植组[(82.5±6.5)、(88.6±6.6)、(77.4±5.6)、(71.2±4.5)],差异有统计学意义(F=51.237、59.432、49.375、51.223,P值均<0.01)。结论PKC-β抑制剂通过减轻炎症浸润及产生保护因子来减轻肾移植后移植物损伤,与MRI检测结果呈对应关系。

Objective To observe the effect of protein kinase C-β(PKC-β)inhibitor on the inflammation in rat transplantation and the corresponding relationship with magnetic resonance imaging(MRI)in grafts.Methods The rats were randomly divided into three groups:isograft group,allograft group and allograft with PKC-βinhibitor treatment group(PKC group),n=10 each.MRI was performed at 24 h after the transplantation,and then the animals were sacrificed.Blood samples were taken to detect the mRNA expression of hypoxia inducible factor-1(HIF-1)and heme oxygenase-1(HO-1)by reverse transcription-polymerase chain reaction(RT-PCR).Immunohistochemistry(IHC)was used to analyze the expression of interleukin-4(IL-4),IL-8,monocyte chemoattractant protein-1(MCP-1)and monocyte migration inhibitory factor(MIF)in kidney tissue.One-way ANOVA test was used for comparisons between groups.Results MRI showed that the arterial spin labeling(ASL)and T1-weighted imaging of rat grafts at 24 h after kidney transplantation in the PKC group were(294.19±42.13)ml/(min×100 g),(1951.1±82.9)ms]respectively,which were significantly higher than in the allograft group[(253.24±25.18)ml/(min×100 g),(1842.1±79.6)ms)]and lower than in the isograft group[(331.65±49.44)ml/(min×100 g),(2047.6±89.3)ms,F=42.323,95.445,P<0.01].The levels of HIF-1 and HO-1 in the PKC group[(1245.17±305.96),(249.72±51.01)]were higher than those in the allograft group[(947.34±112.65),(111.71±41.02)]and the isograft group[(1157.74±224.63),(147.52±42.22),F=61.252,37.832,P<0.01].The expression of IL-4,IL-8,MCP-1,MIF in the grafts of the PKC group[(45.3±4.3),(47.2±4.5),(37.7±3.6),(31.2±3.0)]was significantly higher than the allograft group[(19.4±2.6),(19.6±2.5),(17.4±2.1),(16.7±2.5)],but lower than the isograft group[(82.5±6.5),(88.6±6.6),(77.4±5.6),(71.2±4.5),F=51.237,59.432,49.375,51.223,P<0.01].Conclusion PKC-βinhibitor can reduce the graft damage after kidney transplantation by reducing inflammation and producing protective factors,which is in a corresponding relationship with the results of MRI.