新风胶囊通过调控凋亡相关蛋白促进类风湿关节炎患者外周血CD4~+T细胞凋亡
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摘要
目的:探讨新风胶囊(XFC)对类风湿关节炎患者外周血CD4~+T细胞凋亡的影响。方法:采用随机、双盲法将28例活动期类风湿关节炎患者分为XFC组15例和来氟米特组(LEF组)13例,采用流式细胞术分析CD4~+T细胞凋亡水平,Western blot法和实时定量PCR检测凋亡相关蛋白及mRNA相对表达水平。结果:与治疗前比较,治疗后XFC组外周血红细胞沉降率(ESR)、C-反应蛋白(CRP)、抗环瓜氨酸肽(CCP)抗体、DAS28、B淋巴细胞瘤-2(Bcl-2)mRNA水平明显降低(P <0.01),而CD4~+T细胞凋亡水平、Fas mRNA、FasL mRNA、Caspase-8 mRNA、Caspase-3 mRNA、Bax mRNA和Fas、FasL、Caspase-8、Caspase-3蛋白水平明显升高(P <0.01);与LEF组比较,XFC组ESR、CRP、Bcl-2 m RNA表达水平明显降低(P <0.01),CD4~+T细胞凋亡水平、Fas mRNA、FasL mRNA、Caspase-8mRNA、Caspase-3 mRNA、Bax mRNA和Fas、FasL、Caspase-8、Caspase-3表达水平差异无统计学意义(P> 0.05)。结论:XFC通过上调Fas、Fas L、Caspase-8、Caspase-3促凋亡蛋白表达,下调Bcl-2抑制凋亡蛋白表达,促进异常活化的CD4~+T细胞凋亡,从而抑制类风湿关节炎患者机体的免疫炎症反应改善病情。
Objective:To investigate the effect of Xinfeng Capsule(XFC, 新风胶囊)on apoptosis of CD4~+T cells in peripheral blood of patients with rheumatoid arthritis.Methods:Twenty-eight patients with RA were randomly and double-blind divided into XFC group(15 cases)and LEF group(13 cases).The apoptosis level of CD4~+ T cells was analyzed by flow cytometry.The expression of apoptosis-related proteins and mRNA were detected by Western blot and RT-PCR.Results:Compared with before treatment,the level of ESR,CRP,antiCCP antibodies,DAS28 and Bcl-2mRNA were significantly decreased(P < 0.01),the level of CD4~+T cells apoptosis,(Fas,FasL,Caspase-8, Caspase-3,Bax)mRNA and the proteins expression of Fas,FasL,Caspase-8 and Caspase-3 were significantly increased after XFC treatment(P < 0.01).Compared with LEFgroup,the level of ESR,CRP and Bcl-2 mRNA were significantly decreased in XFC group(P < 0.01),while the level of CD4~+T cells apoptosis(Fas, FasL, Caspase-8,Caspase-3,Bax)mRNA and the proteins expression of Fas,FasL,Caspase-8 and Caspase-3 were significantly increased after XFC treatment have no statistical differences(P > 0.05).Conclusion:XFC can up-regulate the expression of Fas,FasL,Caspase-8 and Caspase-3, down-regulate the expression of Bcl-2, and promoting the apoptosis of CD4~+T cells to inhibit the imm-une inflammatory response of rheumatoid arthritis patients and improving the disease.
Objective:To investigate the effect of Xinfeng Capsule(XFC, 新风胶囊)on apoptosis of CD4~+T cells in peripheral blood of patients with rheumatoid arthritis.Methods:Twenty-eight patients with RA were randomly and double-blind divided into XFC group(15 cases)and LEF group(13 cases).The apoptosis level of CD4~+ T cells was analyzed by flow cytometry.The expression of apoptosis-related proteins and mRNA were detected by Western blot and RT-PCR.Results:Compared with before treatment,the level of ESR,CRP,antiCCP antibodies,DAS28 and Bcl-2mRNA were significantly decreased(P < 0.01),the level of CD4~+T cells apoptosis,(Fas,FasL,Caspase-8, Caspase-3,Bax)mRNA and the proteins expression of Fas,FasL,Caspase-8 and Caspase-3 were significantly increased after XFC treatment(P < 0.01).Compared with LEFgroup,the level of ESR,CRP and Bcl-2 mRNA were significantly decreased in XFC group(P < 0.01),while the level of CD4~+T cells apoptosis(Fas, FasL, Caspase-8,Caspase-3,Bax)mRNA and the proteins expression of Fas,FasL,Caspase-8 and Caspase-3 were significantly increased after XFC treatment have no statistical differences(P > 0.05).Conclusion:XFC can up-regulate the expression of Fas,FasL,Caspase-8 and Caspase-3, down-regulate the expression of Bcl-2, and promoting the apoptosis of CD4~+T cells to inhibit the imm-une inflammatory response of rheumatoid arthritis patients and improving the disease.
引文
[1]端淑杰,刘健,张帆,等.基于关联规则挖掘中医内外合治类风湿关节炎对免疫炎症的影响[J].风湿病与关节炎,2017,6(2):10-14,21.
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[3]ROBERTS CA,DICKINSON AK,TAAMS LS.The Interplay Between Monocytes/Macrophages and CD4~+TCell Subsets in Rheumatiod Arthritis[J].Front Immunol,2015,6(3):571.
[4]梅序桥,吴阿阳,郑燕苹,等.类风湿关节炎患者外周血T淋巴细胞亚群CD4~+/CD8~+表达的意义[J].检验医学与临床,2015,12(21):3144-3146.
[5]谭悦,钱龙,李向培,等.芳香烃受体在类风湿关节炎患者外周血CCR6~+CD4~+T细胞的表达及意义[J].免疫学杂志,2014,30(7):612-616.
[6]黄蓓,汪庆童,刘亢亢,等.类风湿关节炎发生发展中TNF-α信号通路与CD4~+T细胞的关系[J].中国药理学通报,2013,29(7):900-903.
[7]曹永贺,刘健.类风湿性关节炎患者外周血CD4~+T细胞存在凋亡缺陷[J].细胞与分子免疫学杂志,2015,31(5):682-685.
[8]纵瑞凯.新风胶囊组方及其治疗类风湿关节炎的作用[J].风湿病与关节炎,2013,2(1):47-49.
[9]朱福兵,刘健,方利,等.类风湿关节炎患者血小板参数变化可能与BTLA、Treg表达失衡有关[J].风湿病与关节炎,2016,5(7):15-19,23.
[10]王玉辉.类风湿性关节炎患者血清Th17/Treg细胞变化特征[J].北华大学学报(自然科学版),2018,19(1):53-55.
[11]郑杰,王晓元,刘丹,等.艾拉莫德(T-614)对类风湿关节炎患者外周血单个核细胞凋亡和IL-8趋化因子免疫调节作用实验研究[J].陕西医学杂志,2015,44(7):791-793.
[12]朵瑞雪,戎梦瑶,朱平.CsA通过抑制免疫突触的形成促进类风湿关节炎巨噬细胞的凋亡[J].细胞与分子免疫学杂志,2011,27(5):571-573.
[13]王鑫,魏平.类风湿关节炎患者B细胞上Fas受体表达与疾病活动度相关性研究[J].医学研究杂志,2016,45(8):138-141.
[14]俞晨,张俊强,曹威,等.白藜芦醇通过抑制自噬相关蛋白LC3Ⅰ/Ⅱ和beclin-1表达促进H2O2处理的类风湿性关节炎成纤维样滑膜细胞凋亡[J].中国组织化学与细胞化学杂志,2018,27(1):30-33.
[15]李学荣,陈永平,张榜硕,等.艾拉莫德联合依那西普对难治性类风湿关节炎患者CD4~+、CD8~+T细胞及免疫球蛋白的影响[J].免疫学杂志,2018,34(2):141-145.
[16]FAS SC,FRITZSCHING B,SURI-PAYER E,et al.Death receptor signaling and its function in the immune system[J].Curr Dir Autoimmun,2006(9):1-17.
[17]CALMON-HAMATY F,AUDO R,COMBE B,et al.Targeting the Fas/FasL system in rheumatoid arthritis therapy:promising or risky?[J].Cytokine,2015,75(2):228-233.
[18]BLEICKEN S,HANTUSCH A,DAS KK,et al.Quantitative interactome of a membrane Bcl-2 network identifies a hierarchy of complexes for apoptosis regumation[J].Nat Commun,2017,8(1):73.
[19]PU X,STORR SJ,ZHANG Y,et al.Caspase-3 and Caspase-8expression in breast cancer:caspase-3 is associated with survival[J].Apoptosis,2017,22(3):357-368.
[20]罗海恩,毛新展.NF-κB p65基因沉默对类风湿性关节炎滑膜细胞增殖凋亡的影响[J].中国医药导报,2017,14(35):8-11.
[21]曹永贺,刘健.类风湿关节炎患者细胞凋亡与临床指标相关性分析[J].中国中西医结合杂志,2016,36(1):35-39.
[22]FANG WANG,MEI MENG,LI CHEN,et al.Development and validation of a High-performance thin-layer chromatographic fingerprint method for the evaluation of QiYi capsules with the reference of myotonin[J].JPlanar Chromatog,2014,27(3):199-203.
[23]张虹,季有波,杨琪,等.黄芪多糖诱导乳腺癌细胞凋亡的实验研究[J].中国社区医师,2017,33(34):5,7.
[24]曹国春,梁军,侯亚义.薏苡仁油诱导乳腺癌细胞系MCF-7细胞的凋亡及机理研究[J].实用临床医药杂志,2007,11(3):1-3.
[25]叶蕾,贾霜,潘文萍.雷公藤甲素对类风湿关节炎患者滑膜成纤维样细胞增殖的体外抑制作用研究[J].中国药房,2015,26(31):4357-4359.
[26]章平衡,刘健,谈冰,等.新风胶囊通过调节NF-κB通路改善类风湿关节炎患者高凝状态[J].免疫学杂志,2016,32(1):49-55.
[27]刘健,万磊,黄传兵,等.新风胶囊对类风湿关节炎患者脂蛋白代谢的影响[J].中国中西医结合杂志,2015,35(9):1060-1064.
[28]孙玥,刘健,万磊,等.基于B、T淋巴细胞衰减因子及氧化应激探讨新风胶囊改善RA患者心功能机制[C]//全国第十二届中西医结合风湿病学术会议论文汇编,2014:211-212.
[2]MARTENS PB,GORONZY JJ,SCHAID D,et al.Expansion of unusual CD4~+T cells in severe rheumatiod arthritis[J].Arthritis Rheum,2014,40(6):1106-1114.
[3]ROBERTS CA,DICKINSON AK,TAAMS LS.The Interplay Between Monocytes/Macrophages and CD4~+TCell Subsets in Rheumatiod Arthritis[J].Front Immunol,2015,6(3):571.
[4]梅序桥,吴阿阳,郑燕苹,等.类风湿关节炎患者外周血T淋巴细胞亚群CD4~+/CD8~+表达的意义[J].检验医学与临床,2015,12(21):3144-3146.
[5]谭悦,钱龙,李向培,等.芳香烃受体在类风湿关节炎患者外周血CCR6~+CD4~+T细胞的表达及意义[J].免疫学杂志,2014,30(7):612-616.
[6]黄蓓,汪庆童,刘亢亢,等.类风湿关节炎发生发展中TNF-α信号通路与CD4~+T细胞的关系[J].中国药理学通报,2013,29(7):900-903.
[7]曹永贺,刘健.类风湿性关节炎患者外周血CD4~+T细胞存在凋亡缺陷[J].细胞与分子免疫学杂志,2015,31(5):682-685.
[8]纵瑞凯.新风胶囊组方及其治疗类风湿关节炎的作用[J].风湿病与关节炎,2013,2(1):47-49.
[9]朱福兵,刘健,方利,等.类风湿关节炎患者血小板参数变化可能与BTLA、Treg表达失衡有关[J].风湿病与关节炎,2016,5(7):15-19,23.
[10]王玉辉.类风湿性关节炎患者血清Th17/Treg细胞变化特征[J].北华大学学报(自然科学版),2018,19(1):53-55.
[11]郑杰,王晓元,刘丹,等.艾拉莫德(T-614)对类风湿关节炎患者外周血单个核细胞凋亡和IL-8趋化因子免疫调节作用实验研究[J].陕西医学杂志,2015,44(7):791-793.
[12]朵瑞雪,戎梦瑶,朱平.CsA通过抑制免疫突触的形成促进类风湿关节炎巨噬细胞的凋亡[J].细胞与分子免疫学杂志,2011,27(5):571-573.
[13]王鑫,魏平.类风湿关节炎患者B细胞上Fas受体表达与疾病活动度相关性研究[J].医学研究杂志,2016,45(8):138-141.
[14]俞晨,张俊强,曹威,等.白藜芦醇通过抑制自噬相关蛋白LC3Ⅰ/Ⅱ和beclin-1表达促进H2O2处理的类风湿性关节炎成纤维样滑膜细胞凋亡[J].中国组织化学与细胞化学杂志,2018,27(1):30-33.
[15]李学荣,陈永平,张榜硕,等.艾拉莫德联合依那西普对难治性类风湿关节炎患者CD4~+、CD8~+T细胞及免疫球蛋白的影响[J].免疫学杂志,2018,34(2):141-145.
[16]FAS SC,FRITZSCHING B,SURI-PAYER E,et al.Death receptor signaling and its function in the immune system[J].Curr Dir Autoimmun,2006(9):1-17.
[17]CALMON-HAMATY F,AUDO R,COMBE B,et al.Targeting the Fas/FasL system in rheumatoid arthritis therapy:promising or risky?[J].Cytokine,2015,75(2):228-233.
[18]BLEICKEN S,HANTUSCH A,DAS KK,et al.Quantitative interactome of a membrane Bcl-2 network identifies a hierarchy of complexes for apoptosis regumation[J].Nat Commun,2017,8(1):73.
[19]PU X,STORR SJ,ZHANG Y,et al.Caspase-3 and Caspase-8expression in breast cancer:caspase-3 is associated with survival[J].Apoptosis,2017,22(3):357-368.
[20]罗海恩,毛新展.NF-κB p65基因沉默对类风湿性关节炎滑膜细胞增殖凋亡的影响[J].中国医药导报,2017,14(35):8-11.
[21]曹永贺,刘健.类风湿关节炎患者细胞凋亡与临床指标相关性分析[J].中国中西医结合杂志,2016,36(1):35-39.
[22]FANG WANG,MEI MENG,LI CHEN,et al.Development and validation of a High-performance thin-layer chromatographic fingerprint method for the evaluation of QiYi capsules with the reference of myotonin[J].JPlanar Chromatog,2014,27(3):199-203.
[23]张虹,季有波,杨琪,等.黄芪多糖诱导乳腺癌细胞凋亡的实验研究[J].中国社区医师,2017,33(34):5,7.
[24]曹国春,梁军,侯亚义.薏苡仁油诱导乳腺癌细胞系MCF-7细胞的凋亡及机理研究[J].实用临床医药杂志,2007,11(3):1-3.
[25]叶蕾,贾霜,潘文萍.雷公藤甲素对类风湿关节炎患者滑膜成纤维样细胞增殖的体外抑制作用研究[J].中国药房,2015,26(31):4357-4359.
[26]章平衡,刘健,谈冰,等.新风胶囊通过调节NF-κB通路改善类风湿关节炎患者高凝状态[J].免疫学杂志,2016,32(1):49-55.
[27]刘健,万磊,黄传兵,等.新风胶囊对类风湿关节炎患者脂蛋白代谢的影响[J].中国中西医结合杂志,2015,35(9):1060-1064.
[28]孙玥,刘健,万磊,等.基于B、T淋巴细胞衰减因子及氧化应激探讨新风胶囊改善RA患者心功能机制[C]//全国第十二届中西医结合风湿病学术会议论文汇编,2014:211-212.