三物黄芩汤对白塞病模型小鼠IFN-γ、IL-17A水平的影响  

Effect of Sanwu Huangqin Decoction on IFN-γand IL-17A levels in mice with Behcet's disease

作　　者：朴勇洙[1] 孙妲男[2] 邵音 张岩[3] 王炳予 PIAO Yong-zhu;SUN Dan-nan;SHAO Yin;ZHANG Yan;WANG Bing-yu(The First Affiliated Hospital of Heilongjiang University of Traditional Chinese Medicine, Heilongjiang, Harbin, 150040 China;The Second Affiliated Hospital of Heilongjiang University of Traditional Chinese Medicine, Harbin, Heilongjiang, 150001 China;Heilongjiang University of Traditional Chinese Medicine, Heilongjiang, Harbin, 150040 China;Nangang Branch of Heilongjiang College of Traditional Chinese Medicine, Harbin, Heilongjiang, 150006 China)

机构地区：[1]黑龙江中医药大学附属第一医院,黑龙江哈尔滨150040 [2]黑龙江中医药大学附属第二医院,黑龙江哈尔滨150001 [3]黑龙江中医药大学,黑龙江哈尔滨150040 [4]黑龙江省中医药科学院南岗分院,黑龙江哈尔滨150006

出　　处：《海南医学院学报》2020年第7期486-491,共6页Journal of Hainan Medical University

基　　金：黑龙江省自然科学基金(面上项目)(No.H2017064)。

摘　　要：目的:探讨三物黄芩汤对白塞病(BD)模型小鼠IFN-γ、IL-17A水平的影响。方法:收集临床样本并分离单个核细胞,采用RNA-seq高通量测序并鉴别白塞病与健康受试者的差异基因。通过虾原肌球蛋白致敏构建BD小鼠模型并给予三物黄芩汤浓缩液灌胃治疗,ELISA和流式细胞术检测外周血和单个核细胞中IFN-γ、IL-17A的水平,RT-PCR法检测脾脏组织中TBX21基因的表达水平。结果:与健康受试者相比,BD患者共有31个差异基因,其中19个基因表达上调,12个基因表达下调。PPI分析结果显示差异基因中排名前5的基因分别为TBX21、STAT3、IL15、PRF1和IFNG。动物实验结果表明,与空白组相比,模型组外周血及PBMC中IFN-γ、IL-17A的水平明显增高(P<0.01),三物黄芩汤能明显降低小鼠外周血及PBMC中IFN-γ、IL-17A的水平(P<0.05)。RT-PCR结果显示,与空白组相比,模型组脾脏组织中TBX21 mRNA的表达水平明显增高,(P<0.01)。三物黄芩汤能降低小鼠脾脏组织中TBX21 mRNA的表达水平(P<0.05)。结论:三物黄芩汤通过靶向抑制TBX21 mRNA的表达水平,进而抑制INF-γ和IL-17的水平,达到治疗BD的作用。Objective:To investigate the effect of Sanwu Huangqin Decoction on IFN-γand IL-17A levels in mice with Behcet's disease.Methods:Clinical samples of patients with Behcet's disease were collected and mononuclear cells were isolated.High throughput RNA-seq was used to identify the different genes between patients with Behcet's disease and healthy subjects.Mouse model was constructed by shrimp myosin sensitization and treated with Sanwu Huangqin decoction.The levels of IFN-γand IL-17A in peripheral blood and mononuclear cells were detected by ELISA and flow cytometry.The expression level of TBX21 gene in spleen was detected by RT-PCR.Results:Compared with healthy subjects,31 differential genes were identified in patients with Behcet's disease,of which 19 genes were up-regulated and 12 genes were down-regulated.PPI analysis showed that the top 5 genes were TBX21,STAT3,IL15,PRF1 and IFNG.The results of animal experiments showed that the levels of IFN-γand IL-17A in the peripheral blood and PBMC of the model group were significantly higher compared with the control group(P<0.01).Sanwu Huangqin decoction could significantly reduce the levels of IFN-γand IL-17A in the peripheral blood and PBMC of mice(P<0.05).RT-PCR results showed that the expression of TBX21 mRNA in spleen tissue of model group was significantly higher than that of the control group(P<0.01).Sanwu Huangqin decoction could significantly down regulate the expression of TBX21 mRNA in spleen(P<0.05).Conclusion:Sanwu Huangqin decoction could reduce the levels of IFN-γand IL-17A by inhibiting the expression of TBX21 mRNA,so as to treat Behcet's disease.

关 键 词：三物黄芩汤 白塞病 虾原肌球蛋白 TBX21基因 

分 类 号：R285.5[医药卫生—中药学] R-332[医药卫生—中医学]