Background Dendritic cells(DCs)are the most important antigen-presenting cells due to their professional and extremely efficient antigen-presenting function.The dynamics of cytoskeleton plays crucial regulated roles o...Background Dendritic cells(DCs)are the most important antigen-presenting cells due to their professional and extremely efficient antigen-presenting function.The dynamics of cytoskeleton plays crucial regulated roles on DCs’immune functions and biophysical properties.Several evidences show that tumor-derived suppressive cytokines deteriorate DCs’immune functions through remodeling their F-actin cytoskeleton.But the underlying mechanism is still elusive.Tropomodulin1(Tmod1),a cytoskeleton-binding protein,regulates and stabilizes actin filaments lengths and cytoskeleton architecture,which involves in the regulations of the morphology,formation of neural dendrites and biophysical properties of cells.Our previous studies found that mature DCs(mDCs)had a higher expression of Tmod1 than immature DCs(imDCs). Therefore,it’s hypothesized that Tmod1 maybe involve in the modification of DCs’functions.Objective The aim of the study is to investigate the effects of Tmodl on the immune functions and biophysical properties of DCs and the underlying mechanisms in order to further understand the biological behaviors of DCs.Methods Bone marrow-derived cells were harvested from wild type(C57BL/6 J)mice and Tmod1 knockout mice(Tmod1 overexpressing transgenic(TOT)/Tmod1-/-)and differentiated to immature dendritic cells(imDCs)by rmGM-CSF and rmIL-4.imDCs were then matured by lipopolysaccharides(LPS)treatment.The expressions of the surface markers in DCs,including CD80,CD86,CD40,MHC-Ⅱand CCR7,were detected by flow cytometry,Western blot and qRT-PCR.The inflammation cytokines such as IL-6,IFN-γ,IFN-βand IL-10 were also detected by flow cytometry.The immune functions and the biophysical properties of DCs were compared between the wild type and Tmod1 knockout mice.The F-actin content and dendritic pseudopodia of these two kinds of DCs were detected by flow cytometry and laser scanning confocal microscope respectively.Finally,we detected the MyD88 dependent and independent signaling pathway to discover the molecular mechanisms.R展开更多
Certain diseases are known to cause changes in the physical and biomechanical properties of cells.These include cancer,malaria,and sickle cell anemia among others.Typically,such physical property changes can result in...Certain diseases are known to cause changes in the physical and biomechanical properties of cells.These include cancer,malaria,and sickle cell anemia among others.Typically,such physical property changes can result in several fold increases or decreases in cell stiffness,which are significant and can result in severe pathology and eventual catastrophic breakdown of the bodily functions.While there are developed biochemical and biological assays to detect the onset or presence of diseases,there is always a need to develop more rapid,precise,and sensitive methods to detect and diagnose diseases.Biomechanical property changes can play a significant role in this regard.As such,research into disease biomechanics can not only give us an in-depth knowledge of the mechanisms underlying disease progression,but can also serve as a powerful tool for detection and diagnosis.This article provides some insights into opportunities for how significant changes in cellular mechanical properties during onset or progression of a disease can be utilized as useful means for detection and diagnosis.We will also showcase several technologies that have already been developed to perform such detection and diagnosis.展开更多
目的:观察有氧运动对大鼠脑动脉平滑肌细胞(SMC)的大电导钙激活钾通道(BKCa)生物物理特性的影响,为运动有效预防脑血管疾病的细胞学机制提供实验依据。方法:8周龄雄性Wistar大鼠20只,随机分为两组,安静对照组(SED)和运动组(EX)。运动组...目的:观察有氧运动对大鼠脑动脉平滑肌细胞(SMC)的大电导钙激活钾通道(BKCa)生物物理特性的影响,为运动有效预防脑血管疾病的细胞学机制提供实验依据。方法:8周龄雄性Wistar大鼠20只,随机分为两组,安静对照组(SED)和运动组(EX)。运动组进行12周跑台运动(20 m/min,60 min/d,5 d/w)。12周后,取脑动脉,经酶消化法获得单个脑动脉SMC。采用膜片钳单通道内面向外记录模式,观察各组BKCa通道的生物物理特性。结果:1)本实验中BKCa的单通道电导为221.8±6.1p S(n=8),记录电流反转电位接近0 m V,无整流现象,均符合BKCa单通道特征;2)12周有氧运动使BKCa通道活性提高:钳制电位+40 m V,[Ca2+]i为1μM时,SED和EX组BKCa通道平均开放概率分别为0.28±0.02和0.78±0.07 p S 3)有氧运动使BKCa通道电压依赖性增强:[Ca2+]i为1μM时,SED和EX组开放概率-电压曲线左移,V1/2分别为48.09±3.27 m V和26.57±3.17 m V;4)有氧运动使BKCa通道钙敏感性增强:膜电位+40 m V下,SED和EX组Po-[Ca2+]i曲线左移,Kd值分别为2.80±0.09μM和0.35±0.09μM;5)运动组大鼠BKCa通道平均开放时间(To)增大,平均关闭时间(Tc)缩短,开放时间常数(τO)增大,关闭时间常数(τC)减小,提示有氧运动使通道更易开放且更难关闭。结论:长期规律性有氧运动能改变大鼠SMC的BKCa通道门控特性,使其活性增强,这可能是运动改善脑血管舒缩功能,增强脑动脉供血的重要机制之一。展开更多
基金funded by the National Natural Science Foundation of China ( 31660258,31771014, 31860262,31570938,31260227)the Science and Technology Foundation of Guizhou Province ( 2019-2787,2018-1412, 2016-5676,2017-5718)+2 种基金the Science and Technology Innovative Talent Team of Guizhou Province ( 2015-4021)the 2011 Collaborative Innovation Program of Guizhou Province ( 2015-04 )the Cell and Gene Engineering Innovative Research Groups of Guizhou Province ( KY-2016-031)
文摘Background Dendritic cells(DCs)are the most important antigen-presenting cells due to their professional and extremely efficient antigen-presenting function.The dynamics of cytoskeleton plays crucial regulated roles on DCs’immune functions and biophysical properties.Several evidences show that tumor-derived suppressive cytokines deteriorate DCs’immune functions through remodeling their F-actin cytoskeleton.But the underlying mechanism is still elusive.Tropomodulin1(Tmod1),a cytoskeleton-binding protein,regulates and stabilizes actin filaments lengths and cytoskeleton architecture,which involves in the regulations of the morphology,formation of neural dendrites and biophysical properties of cells.Our previous studies found that mature DCs(mDCs)had a higher expression of Tmod1 than immature DCs(imDCs). Therefore,it’s hypothesized that Tmod1 maybe involve in the modification of DCs’functions.Objective The aim of the study is to investigate the effects of Tmodl on the immune functions and biophysical properties of DCs and the underlying mechanisms in order to further understand the biological behaviors of DCs.Methods Bone marrow-derived cells were harvested from wild type(C57BL/6 J)mice and Tmod1 knockout mice(Tmod1 overexpressing transgenic(TOT)/Tmod1-/-)and differentiated to immature dendritic cells(imDCs)by rmGM-CSF and rmIL-4.imDCs were then matured by lipopolysaccharides(LPS)treatment.The expressions of the surface markers in DCs,including CD80,CD86,CD40,MHC-Ⅱand CCR7,were detected by flow cytometry,Western blot and qRT-PCR.The inflammation cytokines such as IL-6,IFN-γ,IFN-βand IL-10 were also detected by flow cytometry.The immune functions and the biophysical properties of DCs were compared between the wild type and Tmod1 knockout mice.The F-actin content and dendritic pseudopodia of these two kinds of DCs were detected by flow cytometry and laser scanning confocal microscope respectively.Finally,we detected the MyD88 dependent and independent signaling pathway to discover the molecular mechanisms.R
文摘Certain diseases are known to cause changes in the physical and biomechanical properties of cells.These include cancer,malaria,and sickle cell anemia among others.Typically,such physical property changes can result in several fold increases or decreases in cell stiffness,which are significant and can result in severe pathology and eventual catastrophic breakdown of the bodily functions.While there are developed biochemical and biological assays to detect the onset or presence of diseases,there is always a need to develop more rapid,precise,and sensitive methods to detect and diagnose diseases.Biomechanical property changes can play a significant role in this regard.As such,research into disease biomechanics can not only give us an in-depth knowledge of the mechanisms underlying disease progression,but can also serve as a powerful tool for detection and diagnosis.This article provides some insights into opportunities for how significant changes in cellular mechanical properties during onset or progression of a disease can be utilized as useful means for detection and diagnosis.We will also showcase several technologies that have already been developed to perform such detection and diagnosis.
文摘目的:观察有氧运动对大鼠脑动脉平滑肌细胞(SMC)的大电导钙激活钾通道(BKCa)生物物理特性的影响,为运动有效预防脑血管疾病的细胞学机制提供实验依据。方法:8周龄雄性Wistar大鼠20只,随机分为两组,安静对照组(SED)和运动组(EX)。运动组进行12周跑台运动(20 m/min,60 min/d,5 d/w)。12周后,取脑动脉,经酶消化法获得单个脑动脉SMC。采用膜片钳单通道内面向外记录模式,观察各组BKCa通道的生物物理特性。结果:1)本实验中BKCa的单通道电导为221.8±6.1p S(n=8),记录电流反转电位接近0 m V,无整流现象,均符合BKCa单通道特征;2)12周有氧运动使BKCa通道活性提高:钳制电位+40 m V,[Ca2+]i为1μM时,SED和EX组BKCa通道平均开放概率分别为0.28±0.02和0.78±0.07 p S 3)有氧运动使BKCa通道电压依赖性增强:[Ca2+]i为1μM时,SED和EX组开放概率-电压曲线左移,V1/2分别为48.09±3.27 m V和26.57±3.17 m V;4)有氧运动使BKCa通道钙敏感性增强:膜电位+40 m V下,SED和EX组Po-[Ca2+]i曲线左移,Kd值分别为2.80±0.09μM和0.35±0.09μM;5)运动组大鼠BKCa通道平均开放时间(To)增大,平均关闭时间(Tc)缩短,开放时间常数(τO)增大,关闭时间常数(τC)减小,提示有氧运动使通道更易开放且更难关闭。结论:长期规律性有氧运动能改变大鼠SMC的BKCa通道门控特性,使其活性增强,这可能是运动改善脑血管舒缩功能,增强脑动脉供血的重要机制之一。
