目的探讨果蝇双翅边缘缺刻同源基因(Notch)信号通路在慢性阻塞性肺疾病(COPD)辅助性T细胞1(Helper T cells 1,Th1)和辅助性T细胞2(Helper T cells 2,Th2)失衡中的作用及芪蛭皱肺颗粒的干预机制。方法70只Wistar大鼠随机挑选10只作为空...目的探讨果蝇双翅边缘缺刻同源基因(Notch)信号通路在慢性阻塞性肺疾病(COPD)辅助性T细胞1(Helper T cells 1,Th1)和辅助性T细胞2(Helper T cells 2,Th2)失衡中的作用及芪蛭皱肺颗粒的干预机制。方法70只Wistar大鼠随机挑选10只作为空白对照组,其余大鼠均采用香烟烟雾(CS)联合气管滴注脂多糖(Lipopolysaccharide,LPS)法建立COPD模型,空白对照组及造模组各随机挑选3只大鼠验证造模是否成功。造模结束进行灌胃给药干预,造模组大鼠随机分为模型对照组、阳性对照组(67.5μg·kg^(-1))及芪蛭皱肺颗粒高中低剂量组(3.24、1.62、0.81 g·kg^(-1)),分别给予生理盐水、醋酸地塞米松混悬液、芪蛭皱肺高、中、低剂量混悬液进行灌胃干预,空白对照组同模型对照组,灌胃等体积生理盐水。经28天造模及28天治疗后,采用动物肺功能测试系统检测吸气峰流速(Peak Inspiratory Flow,PIF)和呼气峰流速(Peak Expiratory Flow,PEF),处死大鼠提取肺脏、脾脏、血清及支气管肺泡灌洗液(BALF),苏木素-伊红(HE)染色评价肺组织病理变化,酶联免疫吸附实验法(ELISA)测定血清及BALF中肿瘤坏死因子-α(TNF-α)含量,流式细胞仪检测脾脏Th1/Th2细胞水平,免疫组织化学法(Immunohistochemistry,IHC)及蛋白免疫印迹法(Western blot)检测肺组织Notch1、Hes家族发状分裂相关增强子1(Hes1)、Hey家族发状分裂相关增强子1(Hey1)蛋白水平,实时荧光定量聚合酶链式反应(Real-time PCR,RT-PCR)检测肺组织Notch1、Hes1、Hey1基因表达水平。结果与空白对照组比较,模型对照组大鼠肺功能显著降低(P<0.05),肺组织出现炎性细胞浸润、支气管结构破坏等病变,血清及BALF中TNF-α含量显著升高(P<0.05),脾Th1细胞百分比显著降低(P<0.05),Th2细胞百分比显著升高(P<0.05),肺组织Notch1、Hes1、Hey1蛋白及mRNA表达显著升高(P<0.05),差异均具有统计学意义;与模型对照组比较,各给药组大鼠肺功能显著升高(P<0.05),肺组织病理损伤均有所减轻,血清及BALF中TNF-α含量显著降低(P<0.05),脾Th1细胞百分比显著升高(P<0.05),Th2细胞百分比显著降低(P<0.05),肺组织Notch1、Hes1、Hey1蛋白及mRNA表达显著降低(P<0.05),差异均具有统计学意义。结论芪蛭皱肺颗粒通过抑制Notch信号通路调节Th1/Th2平衡,从而改善COPD大鼠肺功能及病理损伤,影响其免疫功能。展开更多
OBJECTIVE Experimental autoimmune encephalomyelitis(EAE),the classical animal model for multiple sclerosis(MS)is triggered by an impaired balance of T helper(Th)cells and regulatory T(Tregs)cells.Matrine(MAT),a quinol...OBJECTIVE Experimental autoimmune encephalomyelitis(EAE),the classical animal model for multiple sclerosis(MS)is triggered by an impaired balance of T helper(Th)cells and regulatory T(Tregs)cells.Matrine(MAT),a quinolizidine alkaloid derived from the herb Radix Sophorae Flave,has been shown to ameliorate the clinical signs,inflammatory infiltration,demyelination in acute EAE rats.However,whether MAT protect from EAE by adjusting Th and Treg cells response in specific-cellular and molecular level is unknown.METHODS Herein,MAT was tested for its effects on Th1,Th2,Th17 and Treg cells in the spinal cord of EAE mice and splenocyte-extracted from EAE mice with MOG35-55-restimulated,respectively.RESULTS Our findings revealed that MAT significantly inhibit the proliferation of splenocyte,and remarkably down-regulate the differentiation of Th1/Th17 cells with decreased expressions of CD4+IFN-γ+cells and CD4+IL-17+cells in vivo and IL-17,IFN-γ,ROR-γt,T-bet in vitro,meanwhile it dramatically up-regulate the Th2/Treg cells response associated with increased levels of CD4+TGF-β+1cells and CD4+IL-10+cells in vivo and IL-4,IL-10,TGF-β1,Foxp3 and GATA3in vitro.CONCLUSION Considering the effective therapeutic effects of MAT on EAE,it′s worth to find its new values on other autoimmune diseases.展开更多
目的:研究白细胞介素27(interleukin-27,IL-27)对辅助性T细胞1(helper T cell 1,Th1)分化及急性肺损伤的影响与作用机制。方法:通过盲肠结扎穿刺(cecal ligation and puncture,CLP)构建脓毒症急性肺损伤小鼠模型,苏木精-伊红(hematoxyli...目的:研究白细胞介素27(interleukin-27,IL-27)对辅助性T细胞1(helper T cell 1,Th1)分化及急性肺损伤的影响与作用机制。方法:通过盲肠结扎穿刺(cecal ligation and puncture,CLP)构建脓毒症急性肺损伤小鼠模型,苏木精-伊红(hematoxylin-eosin staining,HE)染色评估肺组织病理损伤。采用Western Blot、RT-qPCR检测JAK/STAT通路表达与激活以及T盒转录因子21(T-box transcription factor 21,T-bet)蛋白的表达。结果:IL-27加重脓毒症小鼠肺损伤。细胞实验表明IL-27显著激活JAK2/STAT1通路磷酸化,上调T-bet蛋白(P<0.01)和mRNA水平(P<0.05)。抑制STAT1磷酸化后,T-bet表达显著下调(P<0.01)。结论:IL-27通过JAK2/STAT1通路促进Th1分化,加重脓毒症小鼠急性肺损伤。展开更多
目的研究中药升降散对脓毒症患者血清Th1/Th2失衡及相关调节因子T-bet、GATA-3的干预作用。方法将55例脓毒症患者随机分为常规组(27例)和升降散组(28例),另将健康志愿者9例设为对照组。常规组患者给予西医常规治疗,升降散组患者在西医...目的研究中药升降散对脓毒症患者血清Th1/Th2失衡及相关调节因子T-bet、GATA-3的干预作用。方法将55例脓毒症患者随机分为常规组(27例)和升降散组(28例),另将健康志愿者9例设为对照组。常规组患者给予西医常规治疗,升降散组患者在西医常规治疗的基础上加用升降散100 m L,每日2次口服或鼻饲,对照组患者予口服安慰剂;3组的疗程均为3 d。比较治疗前后3组患者中医证候评分、白细胞计数(WBC)、C反应蛋白(CRP)、降钙素原(PCT)、血清Th1/Th2及其相关转录因子T-bet、GATA-3的水平。结果常规组与升降散组比较,治疗后中医证候评分、WBC、Th1/Th2和T-bet的差异均有统计学意义(P<0.05或P<0.01),2组患者治疗后CRP、PCT和GATA3的差异无统计学意义(P>0.05);2组与对照组比较,除GATA3外,其他各项指标差异均有统计学意义(P<0.01)。结论升降散可以改善脓毒症患者中医临床征候和炎症反应指标,对血清Th1/Th2失衡及其相关调节因子T-bet、GATA-3的水平具有干预作用。展开更多
基金The project supported by National Natural Science Foundation of China(31570357)
文摘OBJECTIVE Experimental autoimmune encephalomyelitis(EAE),the classical animal model for multiple sclerosis(MS)is triggered by an impaired balance of T helper(Th)cells and regulatory T(Tregs)cells.Matrine(MAT),a quinolizidine alkaloid derived from the herb Radix Sophorae Flave,has been shown to ameliorate the clinical signs,inflammatory infiltration,demyelination in acute EAE rats.However,whether MAT protect from EAE by adjusting Th and Treg cells response in specific-cellular and molecular level is unknown.METHODS Herein,MAT was tested for its effects on Th1,Th2,Th17 and Treg cells in the spinal cord of EAE mice and splenocyte-extracted from EAE mice with MOG35-55-restimulated,respectively.RESULTS Our findings revealed that MAT significantly inhibit the proliferation of splenocyte,and remarkably down-regulate the differentiation of Th1/Th17 cells with decreased expressions of CD4+IFN-γ+cells and CD4+IL-17+cells in vivo and IL-17,IFN-γ,ROR-γt,T-bet in vitro,meanwhile it dramatically up-regulate the Th2/Treg cells response associated with increased levels of CD4+TGF-β+1cells and CD4+IL-10+cells in vivo and IL-4,IL-10,TGF-β1,Foxp3 and GATA3in vitro.CONCLUSION Considering the effective therapeutic effects of MAT on EAE,it′s worth to find its new values on other autoimmune diseases.
文摘目的研究中药升降散对脓毒症患者血清Th1/Th2失衡及相关调节因子T-bet、GATA-3的干预作用。方法将55例脓毒症患者随机分为常规组(27例)和升降散组(28例),另将健康志愿者9例设为对照组。常规组患者给予西医常规治疗,升降散组患者在西医常规治疗的基础上加用升降散100 m L,每日2次口服或鼻饲,对照组患者予口服安慰剂;3组的疗程均为3 d。比较治疗前后3组患者中医证候评分、白细胞计数(WBC)、C反应蛋白(CRP)、降钙素原(PCT)、血清Th1/Th2及其相关转录因子T-bet、GATA-3的水平。结果常规组与升降散组比较,治疗后中医证候评分、WBC、Th1/Th2和T-bet的差异均有统计学意义(P<0.05或P<0.01),2组患者治疗后CRP、PCT和GATA3的差异无统计学意义(P>0.05);2组与对照组比较,除GATA3外,其他各项指标差异均有统计学意义(P<0.01)。结论升降散可以改善脓毒症患者中医临床征候和炎症反应指标,对血清Th1/Th2失衡及其相关调节因子T-bet、GATA-3的水平具有干预作用。
