白细胞介素15和骨桥蛋白在大鼠肾移植急性排斥反应早期的表达

目的 研究白细胞介素(IL)15、骨桥蛋白、穿孔素及颗粒酶B在大鼠肾脏移植急性排斥反应(AR)早期的表达变化.方法 以BN大鼠和LEW大鼠分别作为供受体,建立大鼠肾移植动物模型.环孢素A处理组(CsA组):(BN→LEW,术后注射环孢素A);IL-15阻断组:(BN→LEW,术后注射IL-15抗体);AR组:(BN→LEW,术后注射生理盐水);对照组:(LEW→LEW,术后注射生理盐水).对照组6只大鼠,其余组10只大鼠,于移植后1、3、5、7 d取外周血,逆转录一聚合酶链反应(RT-PCR)动态检测骨桥蛋白、IL-15、穿孔素及颗粒酶B mRNA的表达,并作移植肾病理分析.结果 AR组、CsA组及IL-15阻断组肾移植术后IL-15、骨桥蛋白mRNA表达逐渐上调,第5天均达到高峰;对照组均处于低水平.术后第5天,CsA组和IL-15阻断组IL-15 mRNA分别为9685±1440、4346±741均低于AR组(17 022±2153,P<0.01),IL-15阻断组明显低于CsA组(P<0.01);CsA组和IL-15阻断组骨桥蛋白mRNA分别为13 226±1565、19 112±2908均低于AR组(24 663±2449,P<0.01),IL-15阻断组明显高于CsA组(P<0.01).术后第5天IL-15阻断组和CsA组穿孔素和颗粒酶B mRNA表达明显低于AR组(P<0.01).移植肾病理组织检查显示,AR组呈现重度急性排斥反应,IL-15阻断组排斥反应征象轻微.结论 大鼠肾移植AR发生早期骨桥蛋白、IL-15、穿孔素和颗粒酶B mRNA在外周血中均高水平表达,通过阻断IL-15/IL-15受体途径可明显下调颗粒酶B和穿孔素mRNA的表达.

Expressions of interleukin-15 and osteopontin mRNA in early stage of acute rejection of renal allograft in rats

Objective To study the expressions of interleukin-15(IL-15),osteopontin(OPN), granzyme B(GraB)and perforin (PFP) mRNA in early stage of acute rejection (AR) of renal allograft in rats.Methods The rat renal transplantation model was established.Male Brown Norway and Lewis rats were used as donors and recipients.Four groups were designated:CsA group(BN→LEW,n=10, recipients were treated with CsA i.P.):AB group(BN→LEW,n=10,recipients were treated with anti-IL-15 neutralizing antibody i.P.):AR group(BN→LEW,n=10,recipients were treated with normal saline i.P.)and control group(LEW→LEW,n=6,recipients were treated with normal saline i.P.).The blood samples of recipients were drawn at Days l,3,5 and 7 post-transplantation.The serum expressions ot IL-15.OPN,PFP and GraB mRNA of recipients were detected by real-time PCR.AIIograft tissues were analyzed by pathological assays.Results In comparison with other groups,the expressions of OPN,IL-15, PFP and GraB mRNA in AR group were gradually up.regulated and peaked at Day 5.The expressions of IL-l5 mRNA in CsA and AB groups were 9685±1440 and 4346±741 respectively at Day 5 post-operation. It was significantly lower than that in AR group(17 022±2153,P<0.01).The expression of IL-15 mRNA in AB group was signiflcantly lower than that in CsA group(P<0.01).The expressions of OPN mRNA in CsA and AB groups(13 226±1565 vs 19 112 ±2908)were both significantly lower than that in AR group (24 663±2449.P<0.01).But the expression of OPN mRNA in AB group was higher than that in CsA group(P<0.01).At Day 5 post-transplantation,both the expressions of PFP and GraB mRNA in AB and CsA groups was lower than that in AR group(P<0.01).The pathological results showed that severe AR occurred at Day 7 post-transplantation in AR group and whereas the extent of rejection sign relieved in AB group.Conclusion In early stage of AR of renal allograft in rats,the expressions of OPN,IL-15,PFP and GraB mRNA are up-regulated.Blocking IL-15/IL-15R pathway in early stage of AR can down-regulate the expressions of PFP and GraB mRNA.