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Effects of different flow velocities on behavior and TRPA1 expression in the sea cucumber Apostichopus japonicas 被引量:1
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作者 LIN Chenggang LIU Xiaolu +4 位作者 SUN Lina LIU Shilin SUN Jingchun ZHANG Libin YANG Hongsheng 《Journal of Oceanology and Limnology》 SCIE CAS CSCD 2020年第4期1328-1340,I0001-I0030,共43页
A water flow simulation device capable of adjusting flow velocity was designed in flow velocity range of 0–30 cm/s,with which an indoor experiment was conducted to simulate the movement and adhesive behaviors of diff... A water flow simulation device capable of adjusting flow velocity was designed in flow velocity range of 0–30 cm/s,with which an indoor experiment was conducted to simulate the movement and adhesive behaviors of different-sized Apostichopus japonicus under different flow velocities.Observation showed that,in slow flow(~5 cm/s),A.japonicus moved more distance than in still water,and hardly moved in the riptide(~30 cm/s);and the adhesive capacity of A.japonicus was related to the flow velocity and attachment time.A.japonicus were able to attach to the bottom after any attachment time in the slow flow,after 10 s in the medium flow(~15 cm/s),and after 60 s in the riptide(~30 cm/s).In addition,larger A.japonicus were stronger with adhesive ability than smaller ones.The transcriptome data showed that the expression of transient receptor potential cation channel subfamily A,member 1(TRPA1)in the tube feet was increased significantly in a flowing water,but those in the tentacles and tube feet were not significantly changed.Fluorescence in-situ hybridization results showed that TRPA1 was expressed around the watervascular of tentacles,tube feet,body wall,and spines.Therefore,tube feet were important for sea cucumbers to keep themselves stable in relatively swift flow with adhesion ability. 展开更多
关键词 sea cucumber Apostichopus japonicus BEHAVIOR flow velocity transient receptor potential cation channel subfamily A member 1(trpa1)
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TRPA1 Is Expressed in Central But Not in Peripheral Glia
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作者 Vittorio Vellani Carolina Gomis-Perez +4 位作者 Marcello Pinti Massimiliano Prandini Giorgia Pavesi Chiara Giacomoni Marco Caprini 《Journal of Biomedical Science and Engineering》 2016年第11期515-531,共18页
TRPA1 are cation channels expressed in sensory neurons and in several other cell types. This channel is specifically activated by ally isothiocyanate (AITC), the pungent component of mustard oil, as well as by other e... TRPA1 are cation channels expressed in sensory neurons and in several other cell types. This channel is specifically activated by ally isothiocyanate (AITC), the pungent component of mustard oil, as well as by other electrophilic compounds. Although TRPA1 expression in central glia has been reported, its subcellular localization and its expression in peripheral glia have not been investigated before. In this paper we report the molecular and functional expression of TRPA1 in rat cortical astrocytes. Real-time RT-PCR identified low but significant amounts of TRPA1 mRNA in cortical astrocytes while no signal was seen in peripheral glia isolated from dorsal root ganglia (DRG) or in a glial cell line (DITNC-1). Calcium imaging showed AITC-induced signals in astro-cytes while no response in peripheral glia. AITC induced calcium signals in astrocytes in the presence and in the absence of extracellular calcium, suggesting an intracellular localization of TRPA1 channels. Whole cell electrophysiological recordings were performed in astrocytes, in peripheral glia and in DITNC-1 cells transfected with TRPA1 during AITC application. In TRPA1-transfected DITNC-1 cells typical TRPA1 currents were recorded with a reversal potential near 0 mV, consistent with the opening of a non-selective cation channel. No such currents were recorded in untransfected DITNC-1 cells, in astrocytes and in peripheral glial cells, where even high concentrations of AITC (up to 10 mM) induced no significant outward current. In astrocytes AITC transiently induced an outward rectifying current with the reversal potential near ?90 mV, consistent with K channel activation, likely activated by intracellular release of calcium. Our results suggest that TRPA1 channels are molecularly and functionally expressed in calcium-containing organelles of rat cortical astrocytes, with no expression in the plasma membrane. 展开更多
关键词 ASTROCYTES Intracellular trpa1 DITNC-1 Ensheathing Schwann Cells
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基于TRPA1调控泡沫细胞胆固醇代谢作用探讨枳实薤白桂枝汤治疗AS的作用机制
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作者 何湛湛 杨桢 +9 位作者 陶旭光 陈香云 丁薇 褚策 袁雨露 张雨欣 许咏琪 赵培彰 赵红霞 汪文来 《中国实验方剂学杂志》 CAS CSCD 北大核心 2024年第10期1-10,共10页
目的:基于主动脉瞬时受体电位通道锚蛋白1(TRPA1)调控泡沫细胞胆固醇代谢作用探讨枳实薤白桂枝汤对动脉粥样硬化(AS)小鼠疾病进展的影响与作用机制。方法:采用高脂饲料在载脂蛋白E敲除(ApoE^(-/-))小鼠上建立AS模型,随机分为枳实薤白桂... 目的:基于主动脉瞬时受体电位通道锚蛋白1(TRPA1)调控泡沫细胞胆固醇代谢作用探讨枳实薤白桂枝汤对动脉粥样硬化(AS)小鼠疾病进展的影响与作用机制。方法:采用高脂饲料在载脂蛋白E敲除(ApoE^(-/-))小鼠上建立AS模型,随机分为枳实薤白桂枝汤低、中、高剂量组(2.97、5.94、11.88 g·kg^(-1))与辛伐他汀组(0.002 g·kg^(-1)),每组织喂食高脂饲料同时给药,C57BL/6J小鼠作为正常组使用普通饲料喂养,每组10只。上述过程结束后,取小鼠主动脉与血清。采用苏木素-伊红(HE)染色法观察主动脉根部病理变化;大体油红观察主动脉脂质斑块情况;检测血清中总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)水平,并用酶联免疫吸附测定法(ELISA)检测白细胞介素-1β(IL-1β)、IL-18水平;蛋白免疫印迹法(Western blot)与免疫组化分析TRPA1、三磷酸腺苷(ATP)结合盒转运体A1(ABCA1)、ATP结合盒转运体G1(ABCG1)及甘露糖受体(CD206)表达情况。结果:从药效上看,与正常组比较,模型组主动脉出现斑块、大量泡沫细胞与胆固醇晶体等病理变化,血清TC、LDL-C、IL-1β、IL-18水平均显著升高(P<0.01),HDL-C水平显著下降(P<0.01),主动脉组织CD206水平显著下降(P<0.01);与模型组比较,各给药组均能改善主动脉上的脂质沉积状况,并且除枳实薤白桂枝汤高剂量组外,各给药组均能显著降低TC、LDL-C水平(P<0.01),炎症方面,除枳实薤白桂枝汤中剂量组外,各给药组的IL-1β、IL-18水平均明显降低(P<0.05),此外,枳实薤白桂枝汤可上调CD206水平,且枳实薤白桂枝汤中、高剂量组差异明显(P<0.05)。从作用机制上看,与正常组比较,模型组的主动脉TRPA1、ABCA1、ABCG1表达水平均明显降低(P<0.05);与模型组比较,各给药组均明显提高了主动脉TRPA1表达量(P<0.05),可提高ABCA1、ABCG1的表达量,其中枳实薤白桂枝汤中、高剂量组和辛伐他汀组差异明显(P<0.05),免疫组化结果的趋势与其基本一致。结论:枳实薤白桂枝汤可有效降低血脂与炎症水平,抑制主动脉斑块的形成,其作用机制可能是通过TRPA1引发下游的ABCA1、ABCG1表达升高,促进泡沫细胞中胆固醇流出,从而调控胆固醇代谢并在一定程度上干预炎症水平,最终达到治疗AS的作用。 展开更多
关键词 枳实薤白桂枝汤 动脉粥样硬化 瞬时受体电位通道锚蛋白1(trpa1) 泡沫细胞
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稳定表达“毒性”瞬时受体势A1通道细胞系的建立(英文)
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作者 王少鹏 马良会 +2 位作者 李际仙 彭志刚 曹婧 《生物化学与生物物理进展》 SCIE CAS CSCD 北大核心 2008年第12期1378-1386,共9页
瞬时受体势A1(TRPA1)是一种对低温敏感的离子通道,除响应温度外,也可被各种刺激性化合物激活,是许多感觉模型的转导通道.建立TRPA1异源表达系统将为药理分析及功能研究提供很大的便利,但是TRPA1的表达会引发细胞毒性,因此构建TRPA1稳定... 瞬时受体势A1(TRPA1)是一种对低温敏感的离子通道,除响应温度外,也可被各种刺激性化合物激活,是许多感觉模型的转导通道.建立TRPA1异源表达系统将为药理分析及功能研究提供很大的便利,但是TRPA1的表达会引发细胞毒性,因此构建TRPA1稳定细胞系一直面临着挑战.在人胚肾细胞(HEK-293)中非调控的表达TRPA1稳定细胞系被成功建立.实验证实,培养至25代以上,该细胞系仍持续表达TRPA1,且细胞的功能检测也进一步验证了该重组TRPA1细胞系的稳定性及特异性.TRPA1-HEK细胞系不但是TRPA1功能性分析的便利工具,而且可应用于高通量药物筛选系统,鉴定TRPA1特异性调节剂. 展开更多
关键词 瞬时受体势A1(trpa1) 稳定细胞系 毒性
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柚皮苷抑制致敏肥大细胞脱颗粒及相关炎症因子表达的效应机制研究
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作者 耿蒙利 东淳 +4 位作者 孙丹 卢亮 卞旭 史亮 赵一丁 《辽宁中医杂志》 CAS 2023年第9期229-232,I0008,共5页
目的探讨柚皮苷(naringin,NG)对小鼠肥大细胞瘤细胞系(P815)脱颗粒及相关炎症因子的影响及对瘙痒相关受体的调控作用。方法采用Compound 48/80刺激P815细胞方法建立细胞模型,细胞增殖与活性检测(CCK8)法筛选柚皮苷药物安全浓度,设空白... 目的探讨柚皮苷(naringin,NG)对小鼠肥大细胞瘤细胞系(P815)脱颗粒及相关炎症因子的影响及对瘙痒相关受体的调控作用。方法采用Compound 48/80刺激P815细胞方法建立细胞模型,细胞增殖与活性检测(CCK8)法筛选柚皮苷药物安全浓度,设空白组、模型组、柚皮苷高、中、低剂量组(31.2μg/mL、15.6μg/mL、7.8μg/mL)。采用甲苯胺蓝染色进行肥大细胞计数;联酶免疫吸附法(ELISA法)检测细胞上清液中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)及组胺含量;采用实时荧光定量PCR法(Real-time PCR)检测细胞沉淀中的瞬时受体电位锚蛋白1(transient receptor potential ankyrin 1,TRPA1)、瞬时受体电位香草酸受体1(transient potential vanilloid receptor1,TRPV1)、组胺能第四受体(histamine-4 receptor,H4R)及mRNA的表达。结果(1)与空白组比较,模型组、柚皮苷高、中、低剂量组(NG-H、NG-M、NG-L)组胺及TNF-α含量显著升高(P<0.05),TRPA1、TRPV1及H4R表达均明显上调(P<0.01),模型组、NG-M、NG-L组细胞脱颗粒数目明显增多(P<0.01);(2)与模型组比较,NG-H、NG-M、NG-L组组胺及TNF-α含量降低(P<0.05),NG-H组、NG-M组TRPA1、TRPV1及H4R表达显著下降(P<0.01),NG-L组TRPA1、TRPV1及H4R表达降低(P<0.05);(3)NG-H组抑制组胺及TNF-α释放的作用优于NG-M、NG-L组(P<0.01,P<0.05),抑制TRPV1及H4R表达的作用优于NG-M组、NG-L组(P<0.05)。结论柚皮苷能够抑制体外Compound 48/80诱导的P815细胞脱颗粒及促炎介质释放,对瘙痒相关受体的表达具有抑制作用。 展开更多
关键词 柚皮苷 肥大细胞 组胺 瘙痒 瞬时受体电位锚蛋白1 瞬时受体电位香草酸受体1
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肉桂醛对0℃冷应激大鼠背根神经节神经细胞胞浆内Ca^(2+)变化及TRPA1和TRPM8 mRNA表达的影响 被引量:5
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作者 康欣欣 唐志鹏 +2 位作者 王立娟 吴中华 刘琳琳 《中国实验方剂学杂志》 CAS CSCD 北大核心 2017年第1期127-133,共7页
目的:观察肉桂醛对0℃冷应激大鼠背根神经节神经细胞胞浆内Ca^(2+)浓度变化,并探讨其对瞬时受体电位通道蛋白A1(TRPA1)和M8(TRPM8)mRNA表达的影响。方法:取大鼠原代背根节神经细胞置于0℃冷应激状态,噻唑蓝(MTT)法检测不同时长冷应激状... 目的:观察肉桂醛对0℃冷应激大鼠背根神经节神经细胞胞浆内Ca^(2+)浓度变化,并探讨其对瞬时受体电位通道蛋白A1(TRPA1)和M8(TRPM8)mRNA表达的影响。方法:取大鼠原代背根节神经细胞置于0℃冷应激状态,噻唑蓝(MTT)法检测不同时长冷应激状态下细胞的活性;加入肉桂醛,采用激光共聚焦显微镜观察细胞内外Ca^(2+)分布的变化;另以实时荧光定量PCR(Real-time PCR)检测其TRPA1,TRPM8 mRNA的表达。结果:不同时长的冷应激状态下,细胞存活度无明显差异;冷应激可降低背根神经节(DRG)神经细胞胞浆内Ca^(2+)浓度(P<0.05),一定量的肉桂醛则能使Ca^(2+)浓度显著升高(P<0.01)。Real-time PCR检测显示,长时间冷应激TRPA1 mRNA表达上调(P<0.05);长时间冷应激TRPM8 mRNA表达明显要高于短时冷应激(P<0.05);适当浓度的肉桂醛使冷应激大鼠DRG神经细胞TRPA1 mRNA表达升高(P<0.05,P<0.01),对TRPM8 mRNA的影响尚不明显。结论:冷应激能影响TRPA1,TRPM8 mRNA的表达,降低胞浆内Ca^(2+)浓度,作用机制可能为肉桂醛通过活化TRPA1通道,升高细胞浆内Ca^(2+)浓度。 展开更多
关键词 冷应激 肉桂醛 背根神经节 Ca^2+ 瞬时受体电位通道蛋白A1 瞬时受体电位通道蛋白M8
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瘙痒介质在AEW诱发的小鼠干皮症模型中的表达 被引量:1
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作者 金锦花 李科岩 +1 位作者 王玉慧 薛富善 《昆明医科大学学报》 CAS 2020年第6期5-10,共6页
目的采用丙酮-乙醚-水(acetone-ether-water, AEW)贯序涂抹法建立小鼠干皮症慢性瘙痒模型,检测皮肤和背根神经节(DRG)中MrgprA3和TRPA1 m RNA的表达,初步探讨MrgprA3和TRPA1在AEW诱发的干皮症慢性瘙痒模型中的作用。方法 9周龄C57BL/6雄... 目的采用丙酮-乙醚-水(acetone-ether-water, AEW)贯序涂抹法建立小鼠干皮症慢性瘙痒模型,检测皮肤和背根神经节(DRG)中MrgprA3和TRPA1 m RNA的表达,初步探讨MrgprA3和TRPA1在AEW诱发的干皮症慢性瘙痒模型中的作用。方法 9周龄C57BL/6雄鼠12只,随机平均分为模型组和对照组,模型组颈背部皮肤涂抹丙酮/乙醚(1:1)和水,连续7 d制造小鼠干皮症慢性瘙痒模型;对照组小鼠涂抹蒸馏水,时间频率与模型组相同。观察小鼠搔抓行为学,皮肤组织学(HE染色)、肥大细胞浸润情况(甲苯胺蓝染色),RT-qPCR分析MrgprA3和TRPA1在颈段背根神经节(DRG)和病损皮肤中m RNA表达水平变化。结果与对照组相比,模型组小鼠的抓挠次数显著多于对照组(P<0.01);模型组小鼠病损部位皮肤水分明显减少,皮肤干燥、脱屑,病理提示表皮增厚明显,真皮层肥大细胞浸润不明显;颈部病损皮肤和颈部脊神经节中MrgprA3和TRPA1m RNA表达均高于对照组(P<0.01)。结论利用丙酮-乙醚和水贯序涂抹法成功建立小鼠干皮症慢性瘙痒模型,MrgprA3和TRPA1参与了AEW慢性瘙痒模型的产生和传导。 展开更多
关键词 干皮症 丙酮-乙醚和水贯序涂抹法 慢性瘙痒 瞬时受体电位通道蛋白A1 MAS相关的G蛋白偶联受体A3
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