急性期川崎病信号转导和转录激活因子3的表达及意义

Significance of the alteration of STAT3 in the acute phase of Kawasaki disease

目的:探讨急性期川崎病IL-6介导信号转导和转录激活因子3的表达及意义。方法:急性期川崎病(KD)患儿64例,感染发热患儿18例,正常同年龄对照42例。采用Western blot检测外周血单个核细胞(PBMCs)IL-6刺激前后总蛋白,核蛋白STAT3,pSTAT3的表达水平。荧光定量PCR检测PBMCs IL-6刺激前后及IL-6R抗体阻断后STAT3 mRNA的表达水平。结果:急性期KD患儿PBMCs用IL-6刺激前、后总蛋白STAT3,pSTAT3表达水平高于正常儿童;核蛋白则为IL-6刺激前KD患儿STAT3,pSTAT3表达与正常儿童无明显差异,IL-6刺激后KD患儿STAT3,pSTAT3表达明显高于正常儿童。IL-6刺激前、后KD患儿STAT3 mRNA水平(1.15±0.19,1.74±0.59)均高于感染发热患儿(1.07±0.21,1.45±0.32)及正常儿童(0.56±0.37,1.03±0.51);KD冠脉损伤组(1.19±0.21,1.81±0.47)STAT3 mRNA高于冠脉未损伤组(1.13±0.29,1.73±0.48)。经IL-6R抗体阻断后,KD患儿STAT3 mRNA水平低于IL-6刺激前、后KD患儿及感染发热组(0.99±0.15)。结论:急性期KD患儿体内存在刺激STAT3表达和活化的因素,体外IL-6刺激能增强STAT3的表达和活化并进入细胞核;阻断实验提示IL-6在KD患儿STAT3过度表达、活化中起主导作用,本研究为用IL-6R阻断剂治疗川崎病提供了实验证据。

Objective：To study IL-6 mediated the signal transducer and activator of transcription 3（STAT3） and phosphorylated STAT3（pSTAT3）,STAT3 mRNA expression in acute phase kawasaki disease pathogenesis.Methods：Sixty-four patients with acute kawasaki disease（KD）,eighteen patients with infectious disease（ID） and forty-two age-matched normal controls（NC）were studied.The level of STAT3,pSTAT3 protein in whole protein,nucleoprotein of peripheral blood mononuclear cells（PBMCs）before and after IL-6 stimulation were analysed with Western blot.Real-time PCR were performed to evaluate the level of STAT3 mRNA in PBMCs before and after IL-6 stimulation and IL-6R block.Results：Before and after IL-6 stimulation,STAT3,pSTAT3 proteins in whole protein of acute KD PBMCs were higher than NC group.Before IL-6 stimulation,STAT3,pSTAT3 proteins in nucleoprotein of KD were no difference compared with NC group.But after IL-6 stimulation,STAT3,pSTAT3 proteins in nucleoprotein of acute KD were higher than NC group.Before and after IL-6 stimulation,the expression of STAT3 mRNA in acute KD（1.15±0.19,1.74±0.59） were higher than ID group（1.07±0.21,1.45±0.32） and NC group（0.56±0.37,1.03±0.51）,coronary arterial injury group（KDCAL＋;1.19±0.21,1.81±0.47）were higher than no coronary arterial injury group（KDCAL-;1.13±0.29,1.73±0.48;P0.05）.After IL-6R block,STAT3mRNA in KD reduced compare to before and after IL-6 stimulation in KD and ID group（0.99±0.15）.Conclusion：There are some factors existed to promote express and activation of STAT3 in acute phase kawasaki disease.In vitro IL-6 not only boost expression and activation of the signal molecular transcription but also promote to nucleus.Blocking experiment point out that IL-6 may play a leading role in excessive expression and activation of STAT3 in KD,which for IL-6R blockers therapy provides experimental evidence in kawasaki disease.