小檗碱通过补体和凝血级联改善川崎病冠状动脉内皮细胞损伤的研究

目的 探讨小檗碱(berberine,BBR)通过调节补体和凝血级联改善川崎病(Kawasaki disease,KD)冠状动脉内皮细胞损伤的作用.方法 将人冠状动脉内皮细胞(human coronary artery endothelial cell,HCAEC)分为健康对照组、KD组和BBR处理组(各组n=3).健康对照组、KD组分别加入健康儿童和KD患儿的15%血清,BBR处理组加入KD患儿的15%血清后再加入浓度20 mmol/L BBR.采用同位素标记相对和绝对定量技术和液相色谱-串联质谱技术分析和鉴定差异蛋白表达,并对差异蛋白进行GO功能富集分析和KEGG信号通路富集分析;采用We...

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Published in中国当代儿科杂志 Vol. 27; no. 1; pp. 101 - 108
Main Authors 廖锦文, 郭鑫, 梁波, 李许霞, 徐明国
Format Journal Article
LanguageChinese
Published 深圳市龙岗区妇幼保健院/汕头大学医学院龙岗妇幼临床学院,广东 深圳 518172%深圳市龙岗区第三人民医院儿科,广东 深圳 518020 2025
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ISSN1008-8830
DOI10.7499/j.issn.1008-8830.2406075

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Abstract 目的 探讨小檗碱(berberine,BBR)通过调节补体和凝血级联改善川崎病(Kawasaki disease,KD)冠状动脉内皮细胞损伤的作用.方法 将人冠状动脉内皮细胞(human coronary artery endothelial cell,HCAEC)分为健康对照组、KD组和BBR处理组(各组n=3).健康对照组、KD组分别加入健康儿童和KD患儿的15%血清,BBR处理组加入KD患儿的15%血清后再加入浓度20 mmol/L BBR.采用同位素标记相对和绝对定量技术和液相色谱-串联质谱技术分析和鉴定差异蛋白表达,并对差异蛋白进行GO功能富集分析和KEGG信号通路富集分析;采用Western blot法检测差异蛋白表达.结果 KD组与健康对照组之间共鉴定出518个差异蛋白(上调蛋白300个,下调蛋白218个).BBR处理组与KD组之间共鉴定出422个差异蛋白(上调蛋白221个,下调蛋白201个).生物信息学分析结果显示,与健康对照组比较,KD组差异蛋白富集于补体和凝血级联、真核生物的核糖体生物发生;与KD组比较,BBR处理组差异蛋白富集于补体和凝血级联、真核生物的核糖体生物发生.Western blot结果显示,与健康对照组比较,KD组补体C1q亚组分亚基C(complement C1q subcomponent subunit C,C1QC)、激肽原-1(kininogen-1,KNG1)、补体C1s子组件(complement C1s subcomponent,C1S)、C4b结合蛋白α链(C4b-binding protein alpha chain,C4BPA)蛋白表达升高(P<0.05);与KD组比较,BBR处理组KNG1、C1S、C1QC、C4BPA蛋白表达降低(P<0.05).结论 补体和凝血级联可能参与调控BBR治疗KD冠状动脉损伤,C1QC、KNG1、C1S和C4BPA可作为其生物标志物.
AbstractList 目的 探讨小檗碱(berberine,BBR)通过调节补体和凝血级联改善川崎病(Kawasaki disease,KD)冠状动脉内皮细胞损伤的作用.方法 将人冠状动脉内皮细胞(human coronary artery endothelial cell,HCAEC)分为健康对照组、KD组和BBR处理组(各组n=3).健康对照组、KD组分别加入健康儿童和KD患儿的15%血清,BBR处理组加入KD患儿的15%血清后再加入浓度20 mmol/L BBR.采用同位素标记相对和绝对定量技术和液相色谱-串联质谱技术分析和鉴定差异蛋白表达,并对差异蛋白进行GO功能富集分析和KEGG信号通路富集分析;采用Western blot法检测差异蛋白表达.结果 KD组与健康对照组之间共鉴定出518个差异蛋白(上调蛋白300个,下调蛋白218个).BBR处理组与KD组之间共鉴定出422个差异蛋白(上调蛋白221个,下调蛋白201个).生物信息学分析结果显示,与健康对照组比较,KD组差异蛋白富集于补体和凝血级联、真核生物的核糖体生物发生;与KD组比较,BBR处理组差异蛋白富集于补体和凝血级联、真核生物的核糖体生物发生.Western blot结果显示,与健康对照组比较,KD组补体C1q亚组分亚基C(complement C1q subcomponent subunit C,C1QC)、激肽原-1(kininogen-1,KNG1)、补体C1s子组件(complement C1s subcomponent,C1S)、C4b结合蛋白α链(C4b-binding protein alpha chain,C4BPA)蛋白表达升高(P<0.05);与KD组比较,BBR处理组KNG1、C1S、C1QC、C4BPA蛋白表达降低(P<0.05).结论 补体和凝血级联可能参与调控BBR治疗KD冠状动脉损伤,C1QC、KNG1、C1S和C4BPA可作为其生物标志物.
Abstract_FL Objective To explore the role of berberine(BBR)in ameliorating coronary endothelial cell injury in Kawasaki disease(KD)by regulating the complement and coagulation cascade.Methods Human coronary artery endothelial cells(HCAEC)were divided into a healthy control group,a KD group,and a BBR treatment group(n=3 for each group).The healthy control group and KD group were supplemented with 15%serum from healthy children and KD patients,respectively,while the BBR treatment group received 15%serum from KD patients followed by the addition of 20 mmol/L BBR.Differential protein expression was analyzed and identified using isobaric tags for relative and absolute quantitation technology and liquid chromatography-tandem mass spectrometry,followed by GO functional enrichment analysis and KEGG signaling pathway enrichment analysis of the differential proteins.Western blot was used to detect differential protein expression.Results A total of 518 differential proteins were identified between the KD group and the healthy control group(300 upregulated proteins and 218 downregulated proteins).A total of 422 differential proteins were identified between the BBR treatment group and the KD group(221 upregulated proteins and 201 downregulated proteins).Bioinformatics analysis showed that compared to the healthy control group,the differential proteins in the KD group were enriched in the complement and coagulation cascade and ribosome biogenesis in eukaryotes.Compared to the KD group,the differential proteins in the BBR treatment group were also enriched in the complement and coagulation cascade and ribosome biogenesis in eukaryotes.Western blot results indicated that compared to the healthy control group,the expression of complement C1q subcomponent subunit C(C1QC),kininogen-1(KNG1),complement C1s subcomponent(C1S),and C4b-binding protein alpha chain(C4BPA)was increased in the KD group(P<0.05).Compared to the KD group,the expression of KNG1,C1S,C1QC,and C4BPA was decreased in the BBR treatment group(P<0.05).Conclusions The complement and coagulation cascade may be involved in the regulation of BBR treatment for coronary injury in KD,and C1QC,KNG1,C1S,and C4BPA may serve as biomarkers for this treatment.
Author 廖锦文
李许霞
郭鑫
梁波
徐明国
AuthorAffiliation 深圳市龙岗区妇幼保健院/汕头大学医学院龙岗妇幼临床学院,广东 深圳 518172%深圳市龙岗区第三人民医院儿科,广东 深圳 518020
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Author_FL XU Ming-Guo
GUO Xin
LI Xu-Xia
LIAO Jin-Wen
LIANG Bo
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Copyright Copyright © Wanfang Data Co. Ltd. All Rights Reserved.
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DocumentTitle_FL Berberine ameliorates coronary artery endothelial cell injury in Kawasaki disease through complement and coagulation cascades
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Keywords Complement and coagulation cascade
Human coronary artery endothelial cell
Berberine
补体和凝血级联
Kawasaki disease
Coronary artery lesion
川崎病
冠状动脉病变
人冠状动脉内皮细胞
小檗碱
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PublicationTitle 中国当代儿科杂志
PublicationTitle_FL Chinese Journal of Contemporary Pediatrics
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Title 小檗碱通过补体和凝血级联改善川崎病冠状动脉内皮细胞损伤的研究
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