不同治疗方案对川崎病小鼠模型相关细胞因子的作用机制研究  

Study on the Mechanism of Different Treatment Regimens on Related Cytokines in Kawasaki Disease Mouse Model

作　　者：赵欣[1] 赵颖[1] 张雪梅[1] 金晶 ZHAO Xin;ZHAO Ying;ZHANG Xuemei;JIN Jing(Department of Ultrasound,Shaanxi Provincial People’s Hospital,Xi’an Shaanxi 710068,China)

机构地区：[1]陕西省人民医院超声科,陕西西安710068

出　　处：《中国医疗设备》2023年第7期130-138,151,共10页China Medical Devices

基　　金：陕西省国际科技合作计划项目(2021KW-64)。

摘　　要：目的建立川崎病(Kawasaki Disease,KD)小鼠模型,探讨静脉注射免疫球蛋白(Intravenous Immunoglobulin,IVIg)、糖皮质激素(Glucocorticoids,GCs)及人脐带间充质干细胞(Human Umbilical Cord Mesenchymal Stem Cells,hUC-MSCs)对相关因子的作用机制,为KD的治疗提供新的治疗方案。方法按照随机数字方法分为正常组、KD组、IVIg组、GCs组及hUC-MSCs组,均10只,除正常组外其余组小鼠均建立KD模型。建模成功后,IVIg组注射IVIg,GCs组注射甲泼尼龙琥珀酸钠注射液,hUC-MSCs组注射hUC-MSCs 1×10^(6),KD组注射0.9%氯化钠溶液。观察各组小鼠体重,以及高分辨小动物超声仪检查冠脉内径情况;ELISA检测血清凝血酶(Thrombin,TB)、纤维蛋白原(Fibrinogen,FIB)、D-二聚体(D-Dimer,DD)、血栓烷B2(ThromboxaneB2,TXB2)及炎性因子;HE染色及电镜观察组织病变;免疫印记及免疫组化检测核转录因子-κB(Nuclear Transcription Factor Kappa B,NF-κB)及基质金属蛋白酶-9(Matrix Metalloproteinase-9,MMP-9)蛋白及阳性细胞数。结果IVIg组、GCs组及hUC-MSCs组小鼠第1、7、14及28天时体重均高于正常组,冠脉内径厚度均低于KD组(均P<0.05)。与正常组相比,KD组小鼠TB、FIB、DD、TXB2、肿瘤坏死因子-α、白细胞介素(Interleukin,IL)-6及IL-1β升高(P<0.05);与KD组相比,IVIg组、GCs组及hUC-MSCs组小鼠上述指标均显著降低(P<0.05);与GCs组相比,hUC-MSCs组小鼠上述指标均降低(P<0.05)。HE染色及电镜结果显示,正常组小鼠冠状动脉组织结构完好,内弹力膜连续;KD组动脉组织排列紊乱,内皮细胞大量脱落;IVIg组、GCs组及hUCMSCs组小鼠炎症浸润情况减轻,内皮细胞空泡改善。与正常组相比,KD小鼠NF-κB及MMP-9蛋白及阳性细胞数升高存在显著差异(P<0.05);与KD组相比,IVIg组、GCs组及hUC-MSCs组上述指标显著降低(P<0.05);与GCs组相比,hUCMSCs组上述指标显著降低(P<0.05)。结论3种方法治疗川崎病小鼠均可以降低血清TB、FIB、DD及TXB2水平,抗Objective To explore the mechanism of intravenous immunoglobulin(IVIg),glucocorticoids(GCs)and human umbilical cord mesenchymal stem cells(hUC-MSCs)on related cytokines by establishing mouse model of Kawasaki disease(KD),to provide new therapeutic scheme for the treatment of KD.Methods Mice were divided into normal group,KD group,IVIg group,GCs group and hUC-MSCs group according to random number method,with 10 mice in each group.KD model was established in all groups except the normal group.After successful modeling,IVIg group was injected with IVIg,GCs group was injected with methylprednisolone sodium succinate injection,hUC-MSCs group was injected with hUC-MSCs 1×10^(6),and KD group was injected with 0.9%NaCl solution.The weight of mice in each group was observed,and the coronary artery diameter was examined by high-resolution small animal ultrasound.Serum thrombin(TB),fibrinogen(FIB),D-dimer(DD),thromboxane B2(TXB2)and inflammatory factors were detected by ELISA.Tissue lesions were observed by HE staining and electron microscopy.The protein of nuclear transcription factor kappa B(NF-κB)and matrix metalloproteinase-9(MMP-9)and the number of positive cells were detected by immunostaining and immunohistochemistry.Results The body weight of IVIg group,GCs group and hUC-MSCs group was higher than that of normal group at on 1st,7th,14th and 28th day and the inner diameter thickness of coronary artery was lower than that of KD group(all P<0.05).Compared with normal group,KD group mice showed increased TB,FIB,DD,TXB2,tumor necrosis factor-α,interleukin(IL)-6 and IL-1β(P<0.05).Compared with KD group,the above indexes in IVIg group,GCs group and hUC-MSCs group were all significantly decreased(P<0.05).Compared with GCs group,the above indexes in hUCMSCs group significantly were decreased(P<0.05).The results of HE staining and electron microscopy showed that the coronary artery tissue structure was intact and the inner elastic membrane was continuous in normal group;the arrangement of artery tissue was disordered and

关 键 词：川崎病 免疫球蛋白 糖皮质激素 人脐带间充质干细胞 

分 类 号：R54[医药卫生—心血管疾病] Q813[医药卫生—内科学]