摘要
目的探讨核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症小体与下肢静脉性溃疡创面炎症反应的相关性。方法选取24例活动性下肢静脉性溃疡患者(活动性溃疡组),另以24例无渗出性下肢静脉性溃疡患者作为阳性对照(非活动性溃疡组)和24例外伤患者伤口为阴性对照(外伤伤口组)。比较3组患者的临床资料;同时采集创面周围组织标本,采用免疫组化方法检测其NLRP3蛋白表达情况,采用酶联免疫吸附试验(ELISA)检测创面组织中IL-1β和IL-18蛋白水平,RT-PCR法检测创面组织中凋亡相关斑点样蛋白(ASC)、半胱氨酸天冬氨酸蛋白水解酶-1(caspase-1)、C-Jun氨基端激酶(JNK)、丝裂原活化蛋白激酶p38、核因子-κB(NF-κB)p65及NF-κB抑制蛋白α(NF-κB IkBα)的mRNA表达水平,Western blotting法检测创面组织中NLRP3蛋白表达水平。结果非活动性溃疡组和外伤伤口组患者创面炎症反应较活动性溃疡组平缓。活动性溃疡组创面组织中IL-1β和IL-18蛋白表达水平均高于非活动性溃疡组和外伤伤口组[IL-1β:(146.621±11.597)ng/L比(80.967±14.213)ng/L比(84.962±19.484)ng/L,F=136.200,P<0.001;IL-18:(119.814±12.788)ng/L比(72.899±17.220)ng/L比(48.131±10.407)ng/L,F=167.910,P<0.001]。RT-PCR结果提示活动性溃疡组患者创面组织中ASC[(0.030±0.012)ng/L比(0.021±0.005)ng/L比(0.016±0.004)ng/L,F=18.106,P<0.001]、caspase-1[(0.054±0.012)ng/L比(0.013±0.009)ng/L比(0.018±0.006)ng/L,F=130.372,P<0.001]、NF-κB p65[(0.093±0.015)ng/L比(0.038±0.013)ng/L比(0.043±0.014)ng/L,F=110.950,P<0.001]、NF-κB IkB-α[(0.085±0.015)ng/L比(0.078±0.015)ng/L比(0.041±0.016)ng/L,F=53.070,P<0.001]及JNK[(0.075±0.018)ng/L比(0.042±0.013)ng/L比(0.039±0.014)ng/L,F=41.271,P<0.001]的mRNA表达水平均高于非活动性溃疡组和外伤伤口组,而活动性溃疡组创面组织中p38 mRNA表达水平低于非活动性溃疡组[(0.050±0.008)ng/L比(0.064±0.014)ng/L,P<0.05]。Western blotting结果显示活动性溃疡组创面组织中NLRP3蛋白相对表达量高于外伤伤口组及非活动性溃疡组(0.767±0.272比0.605±0.212比0.556±0.183,F=4.804,P=0.012)。结论NLRP3炎症小体与下肢静脉性溃疡创面炎症反应密切相关,可能为下肢静脉性溃疡的治疗提供新的靶点。
Objective To investigate the relationship between the nucleotide binding oligomerization domain like receptor protein 3(NLRP3)inflammasome and inflammatory reaction of venous ulcer of lower extremity.Methods Twenty-four patients with active venous ulcer of lower extremity(active ulcer group),24 patients with non exudative venous ulcer of lower extremity as positive control(non-active ulcer group),and 24 patients with traumatic wound as negative control(traumatic-wound group)were enrolled.The clinical data of the three groups were compared,the tissue samples around the wound were harvested,and the expressions of NLRP3 protein were detected by immunohistochemistry among the three groups.Enzyme linked immunosorbent assay(ELISA)was used to detect the IL-1βand IL-18 protein levels,RT-PCR was used to detect the mRNA expressions of apoptosis associated speck like protein containing CARD(ASC),caspase-1,c-Jun N-terminal kinase(JNK),p38,nuclear factor(NF)-κB p65 and NF-κB inhibitor alpha(NF-κB IkBα),and Western blotting was performed to evaluate the level of NLRP3 inflammasome in wound tissues.Results The inflammatory response in the non-active ulcer group and trauma-wound group were milder than that in the active ulcer group.The levels of IL-1βand IL-18 proteins in the active ulcer group were higher than those in the nonactive ulcer group and the traumatic-wound group[IL-1β:(146.621±11.597)ng/L vs.(80.967±14.213)ng/L vs.(84.962±19.484)ng/L,F=136.200,P<0.001;IL-18:(119.814±12.788)ng/L vs.(72.899±17.220)ng/L vs.(48.131±10.407)ng/L,F=167.910,P<0.001].The results of RT-PCR showed that the mRNA expressions of ASC[(0.030±0.012)ng/L vs.(0.021±0.005)ng/L vs.(0.016±0.004)ng/L,F=18.106,P<0.001],caspase-1[(0.054±0.012)ng/L vs.(0.013±0.009)ng/L vs.(0.018±0.006)ng/L,F=130.372,P<0.001],NF-κB p65[(0.093±0.015)ng/L vs.(0.038±0.013)ng/L vs.(0.043±0.014)ng/L,F=110.950,P<0.001],NF-κB IkB-α[(0.085±0.015)ng/L vs.(0.078±0.015)ng/L vs.(0.041±0.016)ng/L,F=53.070,P<0.001],and JNK[(0.075±0.018)ng/L vs.(0.042±0.013)ng/L vs.(0.039±0.014)ng/L,F=41.271,P<0.001]in the wound tissues of the active ulcer group were higher than those in the non-active ulcer group and the traumatic-wound group.And the mRNA expression of p38 in the wound tissues of the active ulcer group was lower than that in the non-active ulcer group[(0.050±0.008)ng/L vs.(0.064±0.014)ng/L,P<0.05].The result of Western blotting showed that the relative expression level of NLRP3 protein in the wound tissues of the active ulcer group was higher than that in the traumawound group and non-active ulcer group(0.767±0.272 vs.0.605±0.212 vs.0.556±0.183,F=4.804,P=0.012).Conclusion NLRP3 inflammasome is closely related to the wound in venous ulcer of lower extremity and provides a new target to the therapy of venous ulcer of lower extremity.
作者
汪涛
范隆华
陈斌
刘坚军
李旭
陶悦
王晓俊
黎力梦
WANG Tao;FAN Longhua;CHEN Bin;LIU Jianjun;LI Xu;TAO Yue;WANG Xiaojun;LI Limeng(Department of Vascular Surgery,Qingpu Branch of Zhongshan Hospital,Fudan University,Shanghai 201700,P.R.China;Department of Vascular Surgery,Zhongshan Hospital,Fudan University,Shanghai 200032,P.R.China)
出处
《中国普外基础与临床杂志》
CAS
2021年第10期1287-1293,共7页
Chinese Journal of Bases and Clinics In General Surgery
基金
上海市卫生健康委员会青年项目(项目编号:20194Y0286)
复旦大学附属中山医院青浦分院青年项目(项目编号:QY2019-02)
上海市青浦区科委科技发展基金项目(项目编号:QKY2019-14)
上海市青浦区卫健委基础项目(项目编号:W2020-11)
上海市青浦区卫健委面上项目(项目编号:W2019-39)。
关键词
核苷酸结合寡聚化结构域样受体蛋白3炎症小体
下肢静脉性溃疡
炎症反应
nucleotide binding oligomerization domain like receptor protein 3 inflammasome
venous ulcer of lower extremity
inflammatory reactory
