伏隔核瞬时受体电位通道锚蛋白1在小鼠坐骨神经慢性压迫性损伤诱导的神经病理性疼痛中的作用

目的:采用坐骨神经慢性压迫性损伤（chronic constrictive injury, CCI）方法构建神经病理性疼痛小鼠模型,探讨伏隔核（nucleus accumbens, NAc）内瞬时受体电位通道锚蛋白1（transient receptor potential ankyrin 1, TRPA1）对神经病理...     

瞬时感受器电位香草酸受体l在神经病理性疼痛中的作用研究进展

背景 瞬时感受器电位香草酸受体l(transient receptor potential vanilloid 1,TRPV1)是瞬时感受器电位超家族的成员之一,主要表达在初级传入感觉神经元上,是一种非选择性的阳离子通道.近几年对TRPV1的研究取得了较大进展.目的 TRPV1作为一种与痛觉密切相关的离子通道型受体,在神经病理性疼痛中发挥着重要作用.综述 TRPV1在神经病理性疼痛方面的研究进展.内容 TRPV1的结构与分布研究进展,TRPV1在神经损伤后发生表达的变化,以及降低TRPV1的功能后对神经病理性疼痛的影响. 趋向 对TRPV1与神经病理性疼痛的深入研究将为开发新一代镇痛药物提供理论依据.     

缺血性脑损伤与瞬时受体电位M通道的研究进展

缺血性脑血管病是临床常见病、多发病,其发病机制复杂。钙超载在缺血性脑损伤中起重要作用。瞬时受体电位M通道（transient receptor potential melastatin,TRPM）是位于细胞膜上的一类重要的非选择性阳离子通道超家族,对钙离子有较高的通透性,在缺血性脑损伤中起重要作用,对TRPM通道的研究将成为治疗缺血性脑损伤新的靶点。本文就胞内钙离子超载在缺血性脑损伤中的作用、TRPM通道及其参与的缺血性脑损伤的机制予以综述。     

瞬时受体电位通道在膀胱癌中的作用

膀胱癌是一种发病率高,且具有异质性的尿路上皮癌。从浅表膀胱肿瘤到肌层浸润性恶性肿瘤,由于其高频复发及转移进展特征,本身具有难治性。尽管目前已经形成了以手术为主,化疗、放疗、免疫治疗为辅的综合治疗,但化疗方案选择的局限性、放疗的未普及性和免疫治疗的有效率低等问题,使得临床决策依然存在很大瓶颈。近年已有多个靶向治疗在膀胱癌...     

瞬时受体电位阳离子通道8型及瞬时受体电位香草酸亚型1对急性结肠炎小鼠肠道炎症及感觉传导的影响

目的探讨瞬时受体电位香草酸亚型1(TRPV1)及瞬时受体电位阳离子通道8型(TRPM8)蛋白在急性结肠炎小鼠肠道中的表达、相互作用及对炎症和内脏感觉的影响。方法本实验采用30只雄性C57BL/6小鼠采用随机数字表法分为3组, 分别为对照组、模型组、干预组, 每组10只。模型组与干预组小鼠使用质量浓度为30 g/L的DSS共7 d构建结肠炎模型, 同时干预组每天予以0.3%WS-12溶液灌肠, 连续7 d试剂处理。每天同一时间观察记录小鼠生命活力、毛发及体重变化、粪便性状, 测量粪便隐血情况, 进行疾病活动指数(DAI)评分, 并根据病理切片计算组织病理评分;腹壁反射撤退试验检测各组肠道敏感性;酶联免疫吸附试验(ELISA)检测结肠组织炎性因子:白细胞介素(IL)-1β、IL-6、肿瘤坏死因子-α(TNF-α)、缓激肽(BK)活性表达水平;蛋白质印迹法(Western blot)检测结肠组织紧密连接蛋白(ZO-1、Occludin), TRPV1、TRPM8、降钙素基因相关肽α亚单位(CGRP-Gαq)蛋白表达水平。实时定量反转录聚合酶链反应(RT-qPCR)法检测结肠组织炎性因子IL-...     

瞬时受体电位阳离子通道蛋白6在肾脏疾病中的研究进展

瞬时受体电位阳离子通道蛋白6(TRPC6)在细胞钙信号通路传导中发挥了非常重要的作用.TRPC6为重要的足细胞膜蛋白并在维系足细胞正常生物学功能中起到重要作用,TRPC6基因突变在临床上能引起蛋白尿的发生,使肾小球硬化.本文就TRPC6的结构和功能以及与肾脏相关疾病的关系的研究进展作一综述.     

瞬时受体电位通道TRPA1在骨关节炎中的研究进展

瞬时受体电位通道TRPA1是TRP家族中的一种非选择性阳离子通道蛋白,广泛存在于三叉神经、背根、结状神经节中,表达于Aδ和C纤维的初级感觉神经元上。该通道能被低于17℃的伤害性冷刺激、一系列的化学物质刺激及炎症介质激活,产生钙离子内流为主的跨膜电压变化,参与有害性冷刺激的冷觉形成,并具有调控炎症反应、细胞凋亡坏死及介导疼痛的作用。本文就TRPA1在冷刺激致骨关节炎中的作用作一综述。     

瞬时受体电位通道与前列腺疾病

瞬时受体电位(TRP)通道是一类广泛存在于各种细胞的通道蛋白。前列腺癌上皮细胞中TRP离子通道有不同程度的高表达,而慢性前列腺炎中的表达尚不清楚。研究TRP通道在前列腺疾病发病机制中的作用,为疾病的诊断和治疗提供了新的思路。     

Influence of dexamethasone on the expression and distribution of transient receptor potential cation channel 6 in glomerular podocytes

Objective To observe the changes of foot processes, expression and distribution of transient receptor potential cation channel 6 (TRPC6) in podocytes by puromycin aminonucleoside (PAN) and dexamethasone (DEX) intervention, then to investigate the function of TRPC6 in podocytes and its relation to proteinuria in kidney diseases. Methods Podocytes cultured in vitro were divided into three group: control group, PAN stimulation group and DEX intervention group. Mouse podocyte cell line (MPC5) were cultured in 0.02% dimethyl sulfoxide (DMSO) in control group, subjected to PAN (50 μg/ml) treatment alone or with DEX (1 μmol/L) in other two groups for 8 h, 24 h, 48 h. The podocyte morphology was observed and took pictures by phase - contrast microscope, then the differences of morphology and areas were analyzed. The distribution, mRNA expression and protein expression of TRPC6 were detected by indirect immunocyto-fluorescence, real-time quantitative PCR and Western blotting, respectively. Results The well - developed podocyte arborization and interconnection was formed in control group, but PAN led to significant shrinkage of podocytes (P＜0.05), together with podocyte foot process retraction, effacement and loss of cell contact. DEX significantly prevented the shrinkage and apoptosis of podocytes. The apoptosis rate was significantly increased after PAN stimulated 48 h (P＜0.05). Real-time quantitative PCR and Western blotting found TRPC6 mRNA and protein expression were prone to increase in PAN group compared with control group (P＜0.05). The distribution of TRPC6 becamed abnormal in PAN group. DEX decreased TRPC6 mRNA and protein expression at 48 h compared with PAN group (P＜0.05). The abnormal distribution of TRPC6 was also alleviated by the protection of DEX. Conclusion DEX exerts a direct action to podocyte which retains the integrity of slit diaphragm against podocyte injury, and alleviates proteinuria via stabilizing mRNA, protein expression and distribution of TRPC6.     

瞬时受体电位阳离子通道M7在海马神经元损伤中的作用

目的探讨瞬时受体电位阳离子通道M7(TRPM7)在七氟醚预处理缓解缺血缺氧性损伤(OGD)后海马神经元损伤中的作用。方法出生1d的SD大鼠,提取海马神经元,将其随机分为五组:对照组(C组)、七氟醚预处理组(Sev组)、OGD组、七氟醚预处理+OGD组(SD组)和七氟醚预处理+缓激肽(TRPM7特异性激动剂)+OGD(B组)。缺糖缺氧1.5h后复糖复氧,再正常培养24h以制备OGD模型。C组海马神经元仅做正常培养;Sev组海马神经元行2%七氟醚预处理1h;OGD组海马神经元仅制备OGD模型;SD组海马神经元行2%七氟醚预处理1h,24h后制备OGD模型;B组神经元于七氟醚预处理前15 min在培养基中加入缓激肽(TRPM7特异性激动剂,终浓度200μmol/L),之后行2%七氟醚预处理1h,24h后制备OGD模型。正常培养24h后,分别采用MTT法检测神经元相对存活指数,TUNEL凋亡染色法检测神经元凋亡率,Western blot检测TRPM7蛋白含量,实时定量PCR法检测TRPM7 mRNA表达水平,ELISA法测定神经元IL-1β和TNF-α蛋白含量。结果 OGD组海马神经元TRPM7蛋白含量及mRNA表达水平、凋亡率、IL-1β、TNF-αmRNA表达水平及上清蛋白含量明显高于C组(P0.05),而相对存活指数明显降低于C组(P0.05)。SD组海马神经元TRPM7蛋白含量及mRNA表达水平、凋亡率、IL-1β、TNF-αmRNA表达水平及上清蛋白含量明显低于OGD组(P0.05),而相对存活指数明显高于OGD组(P0.05)。B组海马神经元TRPM7蛋白含量及mRNA表达水平、凋亡率、IL-1β、TNF-αmRNA表达水平及上清蛋白含量明显高于SD组(P0.05),而相对存活指数明显低于SD组(P0.05)。结论七氟醚预处理可通过缓解神经元TRPM7过度表达,减轻缺血缺氧性损伤后海马神经元凋亡和炎症反应。     

瞬时受体电位离子通道1拮抗剂对大鼠切口痛模型术后疼痛的影响

目的 观察足底及鞘内注射瞬时受体电位离子通道1(TRPAl)拮抗剂对大鼠切口痛模型术后疼痛的影响.方法 雄性SD大鼠72只,体质量200～250 9,随机分为6组(n=12):空白对照组(C组)、切口痛组(Ⅰ组)、鞘内注射二甲基亚砜组(DMSO1组)、足底注射二甲基亚砜组(DMSO2组)、鞘内注射HC-030031组(H1组)、足底注射HC-030031组(H2组).除空白对照组外,其余各组均行切口痛模型制作.DMSO1组和H1组于术后24 h鞘内分别给予10％ DMSO 20μl和50 μgHC-030031,DMSO2组和H2组于术后24 h足底分别给予10％ DMSO 40μl和100 μg HC-030031.观察各组在给药前1 h(T0),鞘内或足底注射后0.5 h(T1)、1.0 h(T2)、2.0 h(T3)、4.0 h(T4)、6.0h(T5)的机械缩足刺激阈值(PWMT).各组大鼠随机取4只于T4时处死取脊髓腰膨大,Westem blot法检测脊髓TRPA1蛋白表达变化.结果 与C组[(34.4±4.7)g]比较,Ⅰ组[(23.8±4.3)g]、DMSO1组、DMSO2组PWMT明显降低,脊髓TRPA1表达上调(P＜0.05);与DMSO1组[(24.6±3.5)g]、DMS02组[(22.1±4.7)g]比较,H1组[(28.3±2.4)g]和H2组[(29.1±4.2)g]在T4时间点PWMT明显升高,脊髓TRPA1表达下调(P＜0.05).结论 足底或鞘内注射TRPA1拮抗剂可抑制大鼠切口痛模型所致的术后疼痛.     

鞘内注射PDTC对神经病理性痛大鼠TRPM8受体表达的影响

目的探讨NF-κB参与TRPM8受体在大鼠神经病理性痛觉调制中的作用。方法鞘内置管成功的SPF级健康雄性SD大鼠36只,4～6周龄,体重180～200g,采用随机数字表法分为三组:假手术组(S组)、神经病理性痛组(NP组)和NF-κB阻滞剂PDTC组(PDTC组),每组12只。鞘内置管成功后第3天,NP组和PDTC组采用坐骨神经缩窄性损伤(chronic constriction injury,CCI)法制备大鼠神经病理性痛模型;S组只游离坐骨神经不做损伤处理。三组大鼠术后1d开始连续鞘内注射,连续14d(2次/天),PDTC组鞘内注射PDTC 20μg/10μl(20μg PDTC溶于10μl生理盐水),注射完成后10μl生理盐水冲管,S组和NP组分别鞘内注射等容量生理盐水,三组大鼠分别于术前1d、术后1、3、7、10和14d鞘内给药后30min测定冷痛阈、热痛阈和机械痛阈。分别在术后7、14d处死大鼠,采用Western blot法检测背根神经节(DRG)中TRPM8受体和NF-κB p65蛋白含量。结果与S组比较,术后1～14dNP组冷痛阈明显降低、热痛阈明显缩短、机械痛阈明显降低(P0.05);与NP组比较,术后3～14dPDTC组冷痛阈明显增多、热痛阈明显延长、机械痛阈明显升高(P0.05)。与S组比较,术后7、14dNP组TRPM8受体和NF-κB p65蛋白含量明显升高(P0.05);与NP组比较,术后7、14dPDTC组TRPM8受体和NF-κB p65蛋白含量明显降低(P0.05)。结论大鼠DRG中TRPM8和NF-κB p65表达上调参与神经病理性痛的发生发展,抑制NF-κB活化可以减少TRPM8受体的表达上调并且改善大鼠痛觉过敏的症状。     

Lack of association between transient receptor potential cation channel 6 polymorphisms and primary membranous glomerulonephritis

Membranous glomerulonephritis (MGN) is one common cause of idiopathic nephrotic syndrome. Transient receptor potential cation channel 6 (TRPC6) has been identified as causing a familial form of progressive focal and segmental glomerulosclerosis. The objective was to clarify the relationship between TRPC6 polymorphisms and MGN. We recruited a cohort of 134 biopsy-diagnosed MGN patients and 265 healthy subjects. Genotyping of TRPC6 polymorphisms was performed using allele-specific polymerase chain reaction methods. We then analyzed associations between TRPC6 gene polymorphisms and clinical manifestations and pathogenesis of MGN. There was no statistically significant difference of TRPC6 gene rs3824935 C/T, rs17096918 C/T, and rs4326755 A/G polymorphisms between controls and patients with MGN. There was no statistical significance of allele frequencies in these two groups. The characteristics of clinical parameters in TRPC6 gene (rs3284935) C/T polymorphism revealed no difference except proteinuria (p < 0.0005) between CC and non-CC genotype in MGN patients. Besides, no apparent statistically significant differences of rs17096918 C/T (TT and non-TT) and rs4326755 A/G (AA and non-AA) polymorphisms between genotypes were found in the clinical parameters. There is no different genotype distribution between normal controls and patients with MGN of TRPC6 gene. The data also show that TRPC6 gene may not be associated with disease clinical course of MGN.