川崎病患儿外周血单个核细胞基质细胞衍生因子-1和其受体的变化及意义

目的：观察川崎病(KD)患者血浆基质细胞衍生因子1(SDF1)水平及外周血单个核细胞(PBMC)中SDF1的受体CXCR4mRNA表达变化,并分析其与KD及冠状动脉损害的相关关系。方法：采用ELISA方法及荧光定量PCR技术分别测定12例合并冠脉损害和44例无冠脉损害KD患者的急性期与缓解期血浆SDF1浓度及PBMC中CXCR4mRNA表达变化,并与60例正常人比较。结果：KD患者急性期血浆SDF1蛋白水平为1833±395ng/L;PBMC中CXCR4mRNA表达水平为6.57±2.81较对照组升高(P<0.05,P<0.01);缓解期血浆SDF1蛋白和CXCR4mRNA表达水平较急性期下降(P<0.01),但仍显著高于对照组。KD患者合并冠脉损害者PBMC中CXCR4mRNA表达为8.19±2.39,显著高于无冠脉损害者的6.13±2.77(P<0.01)。结论：KD患者血浆SDF1水平及PBMC中CXCR4mRNA表达是增高的,有可能作为判断KD病情活动状态及冠脉损害的免疫学指标。     

核因子-kB信号途径活化对川崎病冠状动脉损害作用的研究

目的 探讨核因子-kB(NF-kB)信号途径在川崎病(KD)冠状动脉免疫损害中的作用.方法 2004年10月至2005年12月武汉市儿童医院收治的KD患儿48例,将患儿分为两组:冠状动脉损害亚组(CALs亚组)11例,无冠状动脉损害亚组(Non-CALs亚组)37例.感染对照组:同年龄组感染发热住院患儿18例.健康对照组:同年龄组健康体检儿童12例.采用免疫印迹(Western blot)法检测外周血单个核细胞(PBMC)中NF-KBp65及IkBα蛋白表达;采用逆转录聚合酶链反应(RT-PCR)检测TNF-α、MCP-lmRNA表达.结果 (1)KD组NF-kBp65明显高于两对照组[(16.53±4.81 vs.(8.37±3.15,0.33±0.05)(P<0.001)].CALs亚组NF-kBp65显著高于Non-CALs亚组[(22.86±3.11)vs.(13.55±3.23)(P<0.05)].KD组NF-kBp65抑制物IKBa明显低于感染对照组及健康对照组[(5.69±2.03)vs.(21.22±4.37,28.58±7.89)(P<0.001)].CALs亚组中胞核NF-KBP65/IkBα比值与CALs的严重程度呈正相关(r=0.536,P<0.05).(2)KD组TNF-αmRNA、MCP-1mRNA较两对照组明显升高[(7.02±2.91)vs.(3.05±2.33,0.61±0.38)(P<0.01);(3.89±1.10)vs.(1.11±0.42.0.04±0.01)(P<0.01)]CALs亚组TNF-αmRNA、MCP-1mRNA明显高于Non-CALs组[(8.79±1.52)vs.(6.13±2.52)(P<0.05);(4.26±0.78)vs.(3.01±0.53)(P<0.05)].结论 NF-kBp65信号通路的活化参与KD急性期血管炎的发生机制,可能参与冠状动脉损害的形成.     

髓样细胞触发受体-1在川崎病发病机制中的作用

目的 探讨髓样细胞触发受体-1(TREM-1)激活在川崎病(KD)发病机制中的作用。方法 45例KD患儿分为有冠脉损害组(CAL组,16例)和无冠脉损害组(NCAL组,29例),同期呼吸道感染患儿作为发热对照组(15例),健康体检儿童作为正常对照组(15例)。采用荧光定量PCR检测外周血单个核细胞(PBMC)中TREM-1 mRNA、DNAX-活化蛋白12 (DAP12) mRNA表达水平,ELISA法检测血清可溶性TREM-1(sTREM-1)、DAP12、单核细胞趋化因子-1(MCP-1)、IL-8的蛋白水平。结果 KD患儿的sTREM-1、DAP12蛋白及TREM-1 mRNA、DAP12 mRNA表达均高于两个对照组 (P < 0.05),且CAL亚组的sTREM-1蛋白及TREM-1 mRNA表达高于NCAL亚组 (P < 0.05)。KD患儿的MCP-1、IL-8高于两对照组 (P < 0.05),且CAL亚组的MCP-1高于NCAL组 (P < 0.05)。KD患儿的sTREM-1与MCP-1呈正相关 (r = 0.523,P < 0.05)。结论 髓样细胞触发受体-1激活可能在KD 发生过程中起重要作用。     

川崎病患儿外周血单个核细胞中γ干扰素诱导蛋白10表达的意义

目的探讨川崎病(KD)患儿外周血单个核细胞(PBMC)中γ干扰素诱导蛋白-10(IP-10) mRNA表达意义。方法实时荧光定量反转录聚合酶链反应(real-time RT-PCR)检测42例KD及40例健康儿童PBMC中IP-10 mRNA表达。结果KD组PBMC中IP-10 mRNA表达水平为0.741±0.161,明显高于健康对照组0.388±0.063(t=12.988 P=0);KD冠状动脉损伤(CAL)亚组PBMC中IP-10 mRNA表达水平为0.825±0.142,高于无冠状动脉损伤(non-CAL)亚组0.708±0.158(t=2.233 P=0.031);静脉注射人免疫球蛋白(IVIG)治疗后IP-10 mRNA表达水平为0.421±0.119,较治疗前0.728±0.170下降(P<0.05),与健康对照组比较无显著性差异(P>0.05);未予IVIG治疗者常规治疗后IP-10 mRNA表达水平为0.674±0.140,较常规治疗前0.780±0.131轻度下降,但无显著性差异(P>0.05)。结论IP-10在KD患儿PBMC中表达增加,参与KD的发病过程,并与KD CAL发生有关,对PBMC中IP-10 mRNA表达抑制可能是IVIG治疗KD的作用机制之一。     

血浆内皮微粒水平变化与川崎病冠状动脉损伤关系的研究

目的 通过观察川崎病患儿血浆内皮微粒(endothelial microparticle,EMP)水平的变化,确定EMP水平变化与川崎病冠状动脉损伤的关系,探讨EMP测定在早期诊断冠状动脉损伤中的价值.方法 30例川崎病患儿均符合日本川崎病研究委员会第4次修订的诊断标准,其中24例为完全型川崎病,6例为不完全型川崎病,并按病程分为急性期、亚急性期、恢复期.根据超声心动图将川崎病组又分为冠状动脉损伤组(6例)和冠状动脉无损伤组(24例).以10例发热伴有皮疹的患儿及10例健康儿童为发热对照组和正常对照组.采用流式细胞术检测血浆中CD31+/CD42b- EMP水平.结果 川崎病患儿急性期血浆EMP水平[(8.18±2.29)％]明显高于恢复期[(2.77±0.85)％]和正常对照组[(1.34±0.38)％](P＜0.01);亚急性期血浆EMP水平[(5.93±1.05)％]明显高于恢复期和正常对照组(P＜0.01);发热对照组血浆EMP水平[(3.66±1.16)％]高于正常对照组(P＜0.05);急性期川崎病患儿中冠状动脉损伤组血浆EMP水平高于冠状动脉无损伤组,差异有统计学意义(P＜0.01).结论 血浆EMP的检测分析有助于川崎病的早期诊断及冠状动脉损伤的早期发现.     

川崎病患儿外周血髓细胞相关蛋白-8/髓细胞相关蛋白-14的表达变化

目的 观察髓细胞相关蛋白-8(MRP-8)/髓细胞相关蛋白-14(MRP-14)异二聚体在川崎病(KD)患儿血清中的表达变化,寻求KD的实验室诊断指标及药物治疗新靶点.方法 选择2009年7月至2010年12月确诊为KD的患儿46例为KD组,根据心脏彩超评定标准又分为冠状动脉扩张组(15例)和冠状动脉未扩张组(31例).选择同期住院有呼吸道感染而无心脏、血液、免疫系统等疾病的发热患儿25例为非KD发热组,选择同期本院门诊体检健康儿童20例为健康对照组.急性期及亚急性期分别采集外周静脉血,采用ELISA法测定血清MRP-8/MRP-14水平,采用半定量RT-PCR法检测白细胞中MRP-8 mRNA和MRP-14 mRNA相对水平.结果 KD组急性期和亚急性期MRP-8/MRP-14水平、MRP-8 mRNA和MRP-14 mRNA相对水平均明显高于非KD发热组和健康对照组(P均＜0.05),非KD发热组与健康对照组比较差异无统计学意义(P＞0.05).KD组急性期MRP-8/MRP-14水平、MRP-8 mRNA和MRP-14 mRNA相对水平均明显高于亚急性期(P均＜0.001).在急性期及亚急性期冠状动脉扩张组血清MRP-8/MRP-14水平及白细胞MRP-8 mRNA和MRP-14 mRNA相对水平均明显高于同期冠状动脉未扩张组(P＜0.05).结论 MRP-8/MRP-14可能参与KD血管炎的病理过程,并参与冠状动脉损害,可作为KD诊断及预测冠状动脉损害的指标之一,可为KD的治疗提供新靶点.     

川崎病患儿血浆可溶性血栓调节蛋白的检测及其意义

目的探讨川崎病(KD)患儿治疗前后血浆可溶性血栓调节蛋白(sTM)含量变化及其临床意义。方法2004-07—2006-07入住南京医科大学附属南京儿童医院的KD患儿67例,将患儿分为无冠状动脉损伤组(39例)和冠状动脉损伤组(28例),按病程分为急性期、亚急性期和恢复期;另以28例健康儿童作为正常对照组。应用酶联免疫吸附法检测血浆sTM水平。结果急性期有、无冠状动脉损伤组血浆sTM质量浓度均明显高于正常对照组(均P<0.01),且冠状动脉损伤组血浆sTM蛋白水平亦显著高于无冠状动脉损伤组(P<0.01);治疗后sTM水平降低,冠状动脉损伤组KD患儿的血浆sTM水平急性期[(74.04±16.68)ng/mL]、亚急性期[(46.48±14.12)ng/mL]和恢复期[(38.94±12.93)ng/mL]均高于健康对照组[(14.00±5.58)ng/mL],差异有显著性(P<0.01);无冠状动脉损伤组KD患儿急性期[(37.06±14.55)ng/mL]、亚急性期[(34.04±12.47)ng/mL]sTM水平亦显著升高,与健康对照组相比差异有显著性(P<0.01),恢复期[(16.42±9.16)ng/mL]与正常组差异无显著性(P>0.05)。结论sTM可能参与了KD冠状动脉损伤的病理过程,检测sTM的水平对评估KD冠状动脉病变具有重要意义。     

CD40L在川崎病发病机制中的作用

目的探讨T淋巴细胞、血小板CD40L在急性期川崎病血管炎及冠状动脉病变发生中的可能作用。方法用流式细胞术双标法,检测23例急性期及IVIG治疗后川崎病(KD)患儿、23例非KD发热患儿T淋巴细胞、血小板CD40L表达的阳性率及平均荧光强度。结果急性期KD组T淋巴细胞表达CD40L阳性细胞率(27.91±5.47)及平均荧光强度(11.74±3.35)明显高于IVIG治疗后组(分别为12.32±1.89%及6.18±1.71%P均<0.01)及发热对照组;同样,急性期KD组血小板CD40L表达的阳性率及平均荧光强度明显高于IVIG治疗后组及发热对照组。结论T淋巴细胞、血小板CD40L在急性期KD患儿中高表达现象,可能在KD急性期血管炎及冠状动脉病变中发挥一定作用。     

川崎病患儿急性期外周血单个核细胞NLRP3炎症小体的表达及其临床意义

目的 探讨Nod样受体蛋白3（NLRP3）炎症小体与川崎病（KD）患儿急性期炎症反应以及冠状动脉损伤的关系。方法 前瞻性纳入2017年1~10月住院的KD患儿42例为研究对象,其中伴冠状动脉损伤（CAL）9例,非冠状动脉损伤（NCAL）33例。另外选取性别、年龄相匹配的15例肺炎发热患儿作为发热对照组,15例健康儿童作为健康对照组。采用实时荧光定量PCR检测外周血单个核细胞NLPR3炎症小体（NLRP3、ASC和caspase-1） mRNA的表达。采用Spearman秩相关分析法评估NLRP3 mRNA表达与血清C反应蛋白（CRP）、红细胞沉降率（ESR）、白细胞介素-6（IL-6）、白细胞介素-1β（IL-1β）、降钙素原、白蛋白及前白蛋白水平的相关性。结果 KD组急性期外周血NLRP3、ASC和caspase-1 mRNA表达明显高于发热对照组及健康对照组（P < 0.05）;CAL组患儿NLRP3 mRNA表达明显高于NCAL组（P < 0.05）。KD患儿急性期NLRP3 mRNA表达与CRP、IL-6、IL-1β、前白蛋白水平存在相关性（r_s分别为0.449、0.376、0.427、-0.416,均P < 0.05）。结论 NLRP3炎症小体可能参与了KD急性期炎症反应及CAL的发生。     

川崎病患儿血清脑利钠肽的水平改变及其与肌钙蛋白Ⅰ的对比分析

目的 通过观察川崎病(KD)患儿脑利钠肽(BNP)的水平变化,并与肌钙蛋白Ⅰ(cTrⅠ)比较,探讨其在KD早期诊断中的作用.方法 选择住院KD患儿21例作为研究对象,20例健康体检儿童作为对照组.对KD组及对照组采用酶联免疫吸附法测定血浆BNP、cTnⅠ浓度,并行心脏多普勒超声检查测定冠状动脉内径.结果 KD患儿血浆BNP水平[(517.26±213.40)μg/L]明显高于对照组[(37.55±7.56)μg/L],差异有非常显著性(P＜0.01);cTnⅠ浓度[(0.31±0.17)μg/L]也高于对照组[(0.13±0.04)μg/L],差异有显著性(P＜0.01).KD患儿BNP异常率(100%)明显高于cTnⅠ(47.7%),差异有非常显著性(P＜0.01);冠状动脉病变组血浆BNP、cTnⅠ水平明显高于非冠状动脉病变组;典型KD患儿及不完全型KD患儿BNP及cTnⅠ血浆水平间差异无显著性(P＞0.05).结论 KD患儿血浆BNP、cTnⅠ浓度均明显升高,而BNP具有更高的特异性及灵敏度.血浆BNP水平测定有助于KD的诊断,尤其是不完全型KD早期诊断.     

川崎病患儿外周血基质金属蛋白酶-9的表达及其与冠状动脉损伤的关系(英文)

目的　观察MMP 9在川崎病 (KD)患儿不同时期表达水平 ,探讨其在冠状动脉损伤中的可能作用。方法　选择KD患儿 2 7例为研究对象 (无冠脉损伤组 10例 ,冠脉损伤组 17例 ) ,取年龄相仿的 10例败血症患儿、10例健康儿童为发热对照组和正常对照组。分别应用明胶酶谱法、酶联免疫吸附法检测血清MMP 9的活性和蛋白浓度 ,半定量RT PCR的方法检测外周血白细胞MMP 9mRNA表达水平。结果　 (1)冠脉损伤组急性期血清MMP 9活性和蛋白水平 (10 .2± 2 .2 ,114 6± 6 91ng/ml)较无冠脉损伤组显著增高 (6 .2± 2 .1,4 5 7± 133ng/ml,P <0 .0 5 )。两组均较正常对照组 (0 .1± 0 .0 ,72± 2 4ng/ml)和发热对照组 (3.1± 1.4 ,2 2 1± 15 4ng/ml)显著增高 (P <0 .0 1或P <0 .0 5 )。 (2 )KD患儿急性期血清MMP 9蛋白水平与外周血白细胞计数显著正相关 (r =0 .4 80 ,P <0 .0 5 )。 (3)两组KD患儿急性期外周血白细胞MMP 9mRNA表达水平无显著性差异 ,但均显著高于发热对照组和正常对照组 (P <0 .0 1)。 (4)川崎病患儿急性期血清MMP 9活性、蛋白浓度和白细胞表达MMP 9mRNA水平 ,在亚急性期、恢复期均依次明显降低 (P <0 .0 1)。结论　MMP 9在川崎病患儿急性期 ,尤其在伴冠状动脉损伤时表达明显升高 ;MMP 9可能参与了川崎病冠状     

川崎病患儿外周血基质金属蛋白酶1的表达及其与冠状动脉损伤的关系

目的观察基质金属蛋白酶1（matrix metalloproteinase-1,MMP-1）在川崎病患儿外周血不同时期的表达水平,探讨其在冠状动脉（简称冠脉）损伤中的作用。方法40例川崎病患儿（无冠脉损伤组23例,冠脉损伤组17例）,按病程分为急性期、亚急性期和恢复期;另分别以10例败血症患儿、10例健康儿童为发热对照组和正常对照组。应用酶联免疫吸附法检测血清MMP-1蛋白水平、逆转录聚合酶链反应（RT-PCR）检测外周血白细胞MMP-1 mRNA表达水平。结果急性期有、无冠脉损伤组血清MMP-1和白细胞表达MMP-1 mRNA水平均明显高于发热对照组和正常对照组（均P〈0．01）,且冠脉损伤组MMP-1蛋白和mRNA水平较无冠脉损伤组升高更为显著（P〈0．05）;到亚急性期、恢复期时MMP-1蛋白和mRNA水平依次明显降低（各期间比较,均P〈0．01）。川崎病组急性期血清MMP-1蛋白水平与外周血白细胞计数呈显著正相关（r=0．750,P〈0．01）。结论MMP-1在川崎病急性期,尤其在冠脉损伤时表达明显升高;MMP-1的过度表达可能与川崎病冠脉损伤的形成有关。     

Soluble forms of the selectin family in children with Kawasaki disease: prediction for coronary artery lesions

AIM: To investigate the relationship between the plasma levels of soluble forms of the selectin family and the incidence of coronary artery lesions (CALs) in patients with Kawasaki disease (KD). METHODS: Thirty-three patients with KD, including group A patients (n = 22) who had no CALs and group B patients (n = 11) who had CALs, as well as age-matched febrile (n = 10) and afebrile controls (n = 11), were studied. RESULTS: Peak plasma E-selectin levels (172.0 +/- 58.6 ng ml(-1)) occurred during the acute phase of KD, while peak plasma P-selectin levels (260.3 +/- 43.2 ng ml(-1)) occurred during the subacute phase of the illness (p<0.05). Plasma L-selectin levels (1757.3 +/- 244.3 ng ml(-1)) during the convalescent phase tended to be higher than in either the acute or the subacute phase (not significant). Before intravenous immunoglobulin treatment, the plasma levels of E- (225.1 +/- 46.8 ng ml(-1)) and P-selectin (259.4 +/- 76.2 ng ml(-1)) of patients with CALs (n = 11) were significantly higher than those of patients (n = 22) with no CALs (E-selectin, 131.6 +/- 36.9 ng ml(-1); P-selectin, 184.9 +/- 84.6 ng ml(-1); p < 0.05). When a plasma E-selectin value before immunoglobulin treatment of >184.7 ng ml(-1) was used as the cut-off point, the sensitivity and specificity for the incidence of CALs were 81.8% and 90.9%, respectively. These findings demonstrate the relationship between plasma levels of selectins and disease severity of Kawasaki vasculitis. CONCLUSION: Higher plasma levels of E-selectin may have potential as a predictor of the incidence of coronary artery lesions in Kawasaki disease patients.     

Induction of MCP1, CCR2, and iNOS expression in THP-1 macrophages by serum of children late after Kawasaki disease

Evidence of premature atherosclerosis late after Kawasaki disease (KD) is accumulating. Given the potential roles of monocyte chemoattractant protein-1 (MCP-1), chemokine receptor CCR-2, and inducible nitric oxide synthase (iNOS) in atherogenesis, we sought to determine whether serum obtained from children late after KD would induce expression of these genes in macrophages in vitro. A total of 79 subjects were studied, which comprised 57 KD patients, 33 of whom had coronary aneurysms, and 22 age-matched controls. Expression of MCP-1, CCR2, and iNOS mRNA in THP-1 macrophages in the presence of patient and control serum was quantified as a ratio to beta-actin mRNA and expressed as a percentage of control. MCP-1 expression was significantly increased in the presence of serum from patients with coronary aneurysms. Expression of CCR2 and iNOS was significantly increased when THP-1 macrophages were incubated with serum from patients with and without coronary aneurysms. The magnitude of induction of MCP-1, CCR2, and iNOS or in combinations correlated positively with serum high-sensitivity C-reactive protein (hs-CRP), and low-density lipoprotein (LDL) cholesterol levels and negatively with high-density lipoprotein (HDL) cholesterol level. In conclusion, the serum of patients with a history of KD induces expression of MCP-1, CCR2, and iNOS in THP-1 macrophages in vitro. Induction of these genes in vivo may be related to chronic inflammation and may have important implications for premature atherosclerosis.     

川崎病患儿CD4+T淋巴细胞表面CD40L表达及其与冠状动脉损伤的关系

目的通过检测川崎病(KD)患儿静脉输注入血丙种球蛋白(丙球)治疗前后外周血T细胞表面CD40L(CD154)表达,探讨KD冠状动脉损伤的发病机制。方法采用流式细胞仪检测26例KD患儿静脉输注丙球治疗前后、16例其他发热性疾病患儿、15例正常儿童外局血T细胞表面的CD40L表达。采用酶联免疫吸附试验检测相应血清中E-选择素。结果KD患儿CD4 T细胞表面CD40L表达及血清E-选择素显著高于其他发热性疾病组及正常对照组(P<0.01),KD患儿静脉输注丙球治疗后明显下降(P<0.01)。CD4 T细胞表面CD40L表达及E-选择素与KD冠状动脉损伤有关,而CD8 T细胞表面CD40L的表达与冠状动脉损伤无关。KD患儿CD4 T细胞表面CD40L表达与E-选择素水平正相关(r=0.626P<0.05)。结论CD40L异常表达及血清E-选择素在KD发病机制中起重要作用。静脉输注丙球能下调CD40L表达及血清E-选择索,有利于血管炎治疗。     

川崎病冠状动脉损伤与纤维蛋白原Bβ-148C/T基因多态性的关系

目的：探讨纤维蛋白原Bβ-148C/T基因多态性与川崎病冠状动脉损伤的关系。方法：收集36例川崎病患儿与49例健康对照儿童的空腹静脉血,用辅助血浆纤维蛋白原活性测定系统测定纤维蛋白原水平和纤维蛋白原的分子功能;采用多聚酶链反应,限制性酶切方法对纤维蛋白原Bβ-148C/T位点的基因型进行测定。结果：川崎病儿童冠状动脉扩张组血浆纤维蛋白原水平明显高于非扩张组及健康对照组(P<0.01); 川崎病儿童冠状动脉扩张组T等位基因频率明显高于健康对照组及非扩张组 (P<0.01)。结论：纤维蛋白原β-148基因多态性及血浆纤维蛋白原水平与川崎病患儿冠状动脉扩张有相关性。［中国当代儿科杂志,2010,12（7）：518-520］     

Association of lower eosinophil-related T helper 2 (Th2) cytokines with coronary artery lesions in Kawasaki disease

Kawasaki disease (KD) is a systemic febrile vasculitis particular coronary artery involvement. Eosinophilia has been found in our and other studies in KD. This study further investigates whether eosinophil-related T helper 2 (Th2) cytokines or the activation marker (eosinophil cationic protein – ECP) is involved in KD with coronary artery lesions (CAL). A total of 95 KD patients were enrolled for this study. Plasma samples were subjected to the measurement of interleukin (IL)-4, IL-5, and eotaxin by Luminex-Bedalyte multiplex beadmates system and to the measurement of ECP by fluoroimmunoassay. Patients with KD had higher eosinophils than controls. Eosinophil-related mediators: IL-4, IL-5, eotaxin, and ECP levels were also higher in KD patients than controls before intravenous immunoglobulin (IVIG) treatment. After IVIG treatment, ECP decreased but IL-4, IL-5, and eotaxin increased significantly. The higher the IL-5 and eosinophil levels after IVIG treatment, the lower rate of CAL was found. Changes of eosinophils after IVIG treatment were positively correlated to changes of IL-5 levels but not ECP levels. An increase of eosinophils and IL-5, but not ECP levels after IVIG treatment, was inversely correlated with CAL formation in KD.