体外艾塞那肽对1型糖尿病患者外周血单个核细胞的免疫调节作用
目的 探讨体外艾塞那肽（Exendin-4）对1型糖尿病（T1DM）患者外周血单个核细胞（PBMC）的免疫调节作用.方法 通过密度梯度离心法分离11例T1DM患者外周血PBMC,体外研究Exendin-4对PBMC的作用.通过Western blotting检测胰高血糖素样肽1受体（GLP-1R）的表达情况;根据检测PBMC中环磷酸腺苷（cAMP）的浓度设置不同Exendin浓度干预组（0、10、100 nmol/L）及拮抗剂组[50 nmol/L Exendin 9（39）＋100 nmol/L Exendin-4];实时荧光定量聚合酶链反应（PCR）和细胞因子微球检测技术分别测定Exend...
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| Published in | 中华糖尿病杂志 Vol. 9; no. 10; pp. 617 - 621 |
|---|---|
| Main Author | |
| Format | Journal Article |
| Language | Chinese |
| Published |
中南大学湘雅二医院内分泌代谢科中南大学糖尿病免疫学教育部重点实验室国家代谢性疾病临床医学研究中心, 长沙,410011
2017
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| Subjects | |
| Online Access | Get full text |
| ISSN | 1674-5809 |
| DOI | 10.3760/cma.j.issn.1674-5809.2017.10.005 |
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| Abstract | 目的 探讨体外艾塞那肽（Exendin-4）对1型糖尿病（T1DM）患者外周血单个核细胞（PBMC）的免疫调节作用.方法 通过密度梯度离心法分离11例T1DM患者外周血PBMC,体外研究Exendin-4对PBMC的作用.通过Western blotting检测胰高血糖素样肽1受体（GLP-1R）的表达情况;根据检测PBMC中环磷酸腺苷（cAMP）的浓度设置不同Exendin浓度干预组（0、10、100 nmol/L）及拮抗剂组[50 nmol/L Exendin 9（39）＋100 nmol/L Exendin-4];实时荧光定量聚合酶链反应（PCR）和细胞因子微球检测技术分别测定Exendin-4干预前后细胞因子[白细胞介素（IL）10、肿瘤坏死因子（TNF）α]在胞内和上清中的变化情况.两组间比较采用t检验,多组间比较采用方差分析.结果（1）T1DM患者PBMC表达GLP-1R,与对照组（0 nmol/L）相比,100 nmol/L Exendin-4干预组GLP-1R表达显著上升（1.97±0.24比1;t=13.89,P〈0.05）,此种效应能够被GLP-1R阻断剂Exendin-9（39）阻断（0.56±0.10比1, t=15.68,P〈0.05）.（2）与对照组相比,1、10、100 nmol/L浓度Exendin-4对T1DM患者PBMC活力无明显影响（F=0.362、0.393、0.374,均P〉0.05）.（3）与对照组相比,10、100 nmol/L Exendin-4上调PBMC中cAMP水平（7.51±0.01比2.56±0.01,17.21±0.04比2.56±0.01,t=19.71、32.89,P〈0.05）.（4）与对照组相比,Exendin-4显著下调PBMC细胞中TNF-α mRNA及上清液中TNF-α水平（F=5.54、5.12,P〈0.05）,此种效应能够被GLP-1R阻断剂Exendin-9（39）阻断（F=5.43、5.97,P〈0.05）;同时Exendin-4可上调上清液中IL-10水平（F=5.95、6.01,P〈0.05）.结论 Exendin-4可上调IL-10的分泌,下调TNF-α的分泌,提示其可能抑制炎症反应. |
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| AbstractList | 目的 探讨体外艾塞那肽（Exendin-4）对1型糖尿病（T1DM）患者外周血单个核细胞（PBMC）的免疫调节作用.方法 通过密度梯度离心法分离11例T1DM患者外周血PBMC,体外研究Exendin-4对PBMC的作用.通过Western blotting检测胰高血糖素样肽1受体（GLP-1R）的表达情况;根据检测PBMC中环磷酸腺苷（cAMP）的浓度设置不同Exendin浓度干预组（0、10、100 nmol/L）及拮抗剂组[50 nmol/L Exendin 9（39）＋100 nmol/L Exendin-4];实时荧光定量聚合酶链反应（PCR）和细胞因子微球检测技术分别测定Exendin-4干预前后细胞因子[白细胞介素（IL）10、肿瘤坏死因子（TNF）α]在胞内和上清中的变化情况.两组间比较采用t检验,多组间比较采用方差分析.结果（1）T1DM患者PBMC表达GLP-1R,与对照组（0 nmol/L）相比,100 nmol/L Exendin-4干预组GLP-1R表达显著上升（1.97±0.24比1;t=13.89,P〈0.05）,此种效应能够被GLP-1R阻断剂Exendin-9（39）阻断（0.56±0.10比1, t=15.68,P〈0.05）.（2）与对照组相比,1、10、100 nmol/L浓度Exendin-4对T1DM患者PBMC活力无明显影响（F=0.362、0.393、0.374,均P〉0.05）.（3）与对照组相比,10、100 nmol/L Exendin-4上调PBMC中cAMP水平（7.51±0.01比2.56±0.01,17.21±0.04比2.56±0.01,t=19.71、32.89,P〈0.05）.（4）与对照组相比,Exendin-4显著下调PBMC细胞中TNF-α mRNA及上清液中TNF-α水平（F=5.54、5.12,P〈0.05）,此种效应能够被GLP-1R阻断剂Exendin-9（39）阻断（F=5.43、5.97,P〈0.05）;同时Exendin-4可上调上清液中IL-10水平（F=5.95、6.01,P〈0.05）.结论 Exendin-4可上调IL-10的分泌,下调TNF-α的分泌,提示其可能抑制炎症反应. 目的 探讨体外艾塞那肽(Exendin-4)对1型糖尿病(T1DM)患者外周血单个核细胞(PBMC)的免疫调节作用.方法 通过密度梯度离心法分离11例T1DM患者外周血PBMC,体外研究Exendin-4对PBMC的作用.通过Western blotting检测胰高血糖素样肽1受体(GLP-1R)的表达情况;根据检测PBMC中环磷酸腺苷(cAMP)的浓度设置不同Exendin浓度干预组(0、10、100 nmol/L)及拮抗剂组[50 nmol/L Exendin 9(39)+100 nmol/L Exendin-4];实时荧光定量聚合酶链反应(PCR)和细胞因子微球检测技术分别测定Exendin-4干预前后细胞因子[白细胞介素(IL)10、肿瘤坏死因子(TNF)α]在胞内和上清中的变化情况.两组间比较采用t检验,多组间比较采用方差分析.结果(1)T1DM患者PBMC表达GLP-1R,与对照组(0 nmol/L)相比,100 nmol/L Exendin-4干预组GLP-1R表达显著上升(1.97±0.24比1;t=13.89,P<0.05),此种效应能够被GLP-1R阻断剂Exendin-9(39)阻断(0.56±0.10比1, t=15.68,P<0.05).(2)与对照组相比,1、10、100 nmol/L浓度Exendin-4对T1DM患者PBMC活力无明显影响(F=0.362、0.393、0.374,均P>0.05).(3)与对照组相比,10、100 nmol/L Exendin-4上调PBMC中cAMP水平(7.51±0.01比2.56±0.01,17.21±0.04比2.56±0.01,t=19.71、32.89,P<0.05).(4)与对照组相比,Exendin-4显著下调PBMC细胞中TNF-α mRNA及上清液中TNF-α水平(F=5.54、5.12,P<0.05),此种效应能够被GLP-1R阻断剂Exendin-9(39)阻断(F=5.43、5.97,P<0.05);同时Exendin-4可上调上清液中IL-10水平(F=5.95、6.01,P<0.05).结论 Exendin-4可上调IL-10的分泌,下调TNF-α的分泌,提示其可能抑制炎症反应. |
| Abstract_FL | Objective To investigate the immunoregulation effects of Exendin-4 on peripheral blood mononuclear cells (PBMC) in patients with type 1 diabetes mellitus (T1DM). Methods PBMC samples from 11 T1DM subjects were used to investigate the effects of Exendin-4 on PBMCs in vitro. Intervention group (0, 10, 100 nmol/L) and antagonist group (50 nmol/L Exendin 9(39)+100 nmol/L Exendin-4)were set according to the concentration of cyclic adenosine monophospphate(cAMP)in PBMCs. Cytokines[interleukin(IL)10,tumor necrosis factor α(TNF-α)]were measured by cytometric bead array and real-time quantitative PCR(RT-qPCR)analysis,respectively.Results (1)GLP-1R was detected in PBMC of T1DM.Compared with control group,the expression of GLP-1R mRNA was significantly increased in 100 nmol/L Exendin-4 treated group (1.97 ± 0.24 vs 1, P<0.05), and was inhibited by GLP-1R antagonist Exendin-9(39)(0.56±0.10 vs 1,P<0.05).(2)Compared with the control group,1 10 100 nmol/L Exendin-4 had no significant effects on PBMC activity in patients with T1DM(F=0.362,0.393,0.374,all P>0.05).(3)10 nmol/L and 100 nmol/L Exendin-4 had significant effects on levels of cAMP(7.51±0.01 vs 2.56±0.01, 17.21±0.04 vs 2.56±0.01,respectively,both P<0.05).(4)The mRNA expression levels of TNF-α,TNF-α levels in the cellular supernatant were significantly decreased after Exendin-4 intervention(F=5.54,5.12,P<0.05),and was inhibited by GLP-1R antagonist Exendin-9(39)(F=5.43,5.97,P<0.05).The levels of IL-10 in supernatant were elevated (F=5.95,6.01, P<0.05) although IL-10 mRNA levels maintained stable in PBMC. Conclusion Exendin-4 could down-regulate TNF-α secretion and up-regulate IL-10 secretion in PBMC of T1DM patients. These results suggest that exendin-4 might down-regulate pro-inflammatory responses. |
| Author | 李霞;雷康;俞海波;段仙兰;彭亚妮;周智广 |
| AuthorAffiliation | 中南大学湘雅二医院内分泌代谢科中南大学糖尿病免疫学教育部重点实验室国家代谢性疾病临床医学研究中心,长沙410011 |
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| Author_FL | Yu Haibo Zhou Zhiguang Li Xia Peng Yani Duan Xianlan Lei Kang |
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| Keywords | 糖尿病,1型 type 1 艾塞那肽 Immunoregulation Diabetes mellitus 胰高血糖素样肽1受体 免疫调节 Exendin-4 Glucagon-like peptide-1 receptor |
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| Notes | Objective To investigate the immunoregulation effects of Exendin-4 on peripheral blood mononuclear cells （PBMC） in patients with type 1 diabetes mellitus （T1DM）. Methods PBMC samples from 11 T1DM subjects were used to investigate the effects of Exendin-4 on PBMCs in vitro. Intervention group （0, 10, 100 nmol/L） and antagonist group （50 nmol/L Exendin 9（39）＋100 nmol/L Exendin-4）were set according to the concentration of cyclic adenosine monophospphate（cAMP）in PBMCs. Cytokines[interleukin（IL）10,tumor necrosis factor α（TNF-α）]were measured by cytometric bead array and real-time quantitative PCR（RT-qPCR）analysis,respectively.Results （1）GLP-1R was detected in PBMC of T1DM.Compared with control group,the expression of GLP-1R mRNA was significantly increased in 100 nmol/L Exendin-4 treated group （1.97 ± 0.24 vs 1, P〈0.05）, and was inhibited by GLP-1R antagonist Exendin-9（39）（0.56±0.10 vs 1,P〈0.05）.（2）Compared with the control group,1 10 100 nmol/L Exendin-4 had no significant effects on PBMC activity in patients wit |
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| PublicationTitle | 中华糖尿病杂志 |
| PublicationTitleAlternate | CHINESE JOURNAL OF DIABETES MELLITUS |
| PublicationTitle_FL | Chinese Journal of Diabetes Mellitus |
| PublicationYear | 2017 |
| Publisher | 中南大学湘雅二医院内分泌代谢科中南大学糖尿病免疫学教育部重点实验室国家代谢性疾病临床医学研究中心, 长沙,410011 |
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| Snippet | 目的 探讨体外艾塞那肽（Exendin-4）对1型糖尿病（T1DM）患者外周血单个核细胞（PBMC）的免疫调节作用.方法 通过密度梯度离心法分离11例T1DM患者外周血PBMC,体外研... 目的 探讨体外艾塞那肽(Exendin-4)对1型糖尿病(T1DM)患者外周血单个核细胞(PBMC)的免疫调节作用.方法 通过密度梯度离心法分离11例T1DM患者外周血PBMC,体外研究Exendin-4... |
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| SubjectTerms | 1型 免疫调节 糖尿病 胰高血糖素样肽1受体 艾塞那肽 |
| Title | 体外艾塞那肽对1型糖尿病患者外周血单个核细胞的免疫调节作用 |
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