The transient receptor potential cation channel subfamily V member 1（TRPV1） provides the sensation of pain（nociception）. However, it remains unknown whether TRPV1 is activated after peripheral nerve injury, or whe...The transient receptor potential cation channel subfamily V member 1（TRPV1） provides the sensation of pain（nociception）. However, it remains unknown whether TRPV1 is activated after peripheral nerve injury, or whether activation of TRPV1 affects neural regeneration. In the present study, we established rat models of unilateral sciatic nerve crush injury, with or without pretreatment with AMG517（300 mg/kg）, a TRPV1 antagonist, injected subcutaneously into the ipsilateral paw 60 minutes before injury. At 1 and 2 weeks after injury, we performed immunofluorescence staining of the sciatic nerve at the center of injury, at 0.3 cm proximal and distal to the injury site, and in the dorsal root ganglia. Our results showed that Wallerian degeneration occurred distal to the injury site, and neurite outgrowth and Schwann cell regeneration occurred proximal to the injury. The number of regenerating myelinated and unmyelinated nerve clusters was greater in the AMG517-pretreated rats than in the vehicle-treated group, most notably 2 weeks after injury. TRPV1 expression in the injured sciatic nerve and ipsilateral dorsal root ganglia was markedly greater than on the contralateral side. Pretreatment with AMG517 blocked this effect. These data indicate that TRPV1 is activated or overexpressed after sciatic nerve crush injury, and that blockade of TRPV1 may accelerate regeneration of the injured sciatic nerve.展开更多
目的:通过观察沉默瞬时受体阳离子通道亚家族V成员6(transient receptor potential cation channel,subfamily V member 6,TRPV6)对结肠癌SW480细胞生物学行为的影响和细胞内钙的浓度变化,以及1,25(OH)_2D_3、CaCl_2及CuCl_2在SD大鼠结...目的:通过观察沉默瞬时受体阳离子通道亚家族V成员6(transient receptor potential cation channel,subfamily V member 6,TRPV6)对结肠癌SW480细胞生物学行为的影响和细胞内钙的浓度变化,以及1,25(OH)_2D_3、CaCl_2及CuCl_2在SD大鼠结肠肿瘤模型建立中的影响,探索TRPV6在结肠癌发生过程中的相关作用,为结肠癌的防治寻找新的靶点。方法:通过构建慢病毒颗粒感染结肠癌SW480细胞,采用免疫组织化学法、Western bolt、PCR技术检测TRPV6蛋白及m RNA的表达,MTT法、迁移及凋亡实验观察结肠癌SW480细胞增殖、迁移及凋亡变化,高速离子成像系统测定结肠癌SW480细胞内的Ca^(2+)浓度变化。以二甲基肼(dimethyl hydrazine,DMH)建立SD大鼠结肠肿瘤模型,分为实验组(DMH组)和干预组[(DMH+1,25(OH)_2D_3组、DMH+CuCl_2组)]、对照组,干预组分别予1,25(OH)_2D_3(37.5 nmol/kg)、CuCl2(375μmol/kg)对模型进行干预。观察各组大鼠结肠腺瘤及腺癌的发生情况,Western blot检测各组结肠组织中TRPV6蛋白的表达情况。结果:TRPV6-RNAi转染结肠癌SW480细胞后,TRPV6 m RNA及蛋白表达减少,细胞Ca^(2+)浓度水平降低,结肠癌SW480细胞的增殖率、迁移能力下降,细胞凋亡率增加,与空白对照组及阴性对照组比较差异具有统计学意义(P<0.05)。对照组大鼠成瘤率为0,DMH+1,25(OH)_2D_3组为100%,DMH组为84.62%,DMH+CuCl_2组为33.33%。各组大鼠大肠中均有TRPV6蛋白的表达,表达情况为DMH+1,25(OH)_2D_3组>DMH组>DMH+CuCl_2组>对照组,差异具有统计学意义(P<0.05)。结论:降低结肠癌SW480细胞Ca^(2+)浓度水平可抑制结肠癌SW480细胞的增殖、迁移能力,诱导细胞凋亡。1,25(OH)_2D_3可以增加实验鼠结肠组织TRPV6蛋白表达,促进结肠肿瘤的形成。CuCl_2可以使实验鼠结肠组织中的TRPV6蛋白表达降低,并抑制结肠肿瘤的形成。展开更多
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
基金supported by the National Natural Science Foundation of China,No.81171178the Natural Science Foundation of Shanxi Province in China,No.2012011036-3Scientific Research Foundation of Shanxi Province of China for the Returned Overseas Chinese Scholars,No.2013011054-2
文摘The transient receptor potential cation channel subfamily V member 1（TRPV1） provides the sensation of pain（nociception）. However, it remains unknown whether TRPV1 is activated after peripheral nerve injury, or whether activation of TRPV1 affects neural regeneration. In the present study, we established rat models of unilateral sciatic nerve crush injury, with or without pretreatment with AMG517（300 mg/kg）, a TRPV1 antagonist, injected subcutaneously into the ipsilateral paw 60 minutes before injury. At 1 and 2 weeks after injury, we performed immunofluorescence staining of the sciatic nerve at the center of injury, at 0.3 cm proximal and distal to the injury site, and in the dorsal root ganglia. Our results showed that Wallerian degeneration occurred distal to the injury site, and neurite outgrowth and Schwann cell regeneration occurred proximal to the injury. The number of regenerating myelinated and unmyelinated nerve clusters was greater in the AMG517-pretreated rats than in the vehicle-treated group, most notably 2 weeks after injury. TRPV1 expression in the injured sciatic nerve and ipsilateral dorsal root ganglia was markedly greater than on the contralateral side. Pretreatment with AMG517 blocked this effect. These data indicate that TRPV1 is activated or overexpressed after sciatic nerve crush injury, and that blockade of TRPV1 may accelerate regeneration of the injured sciatic nerve.
文摘目的:通过观察沉默瞬时受体阳离子通道亚家族V成员6(transient receptor potential cation channel,subfamily V member 6,TRPV6)对结肠癌SW480细胞生物学行为的影响和细胞内钙的浓度变化,以及1,25(OH)_2D_3、CaCl_2及CuCl_2在SD大鼠结肠肿瘤模型建立中的影响,探索TRPV6在结肠癌发生过程中的相关作用,为结肠癌的防治寻找新的靶点。方法:通过构建慢病毒颗粒感染结肠癌SW480细胞,采用免疫组织化学法、Western bolt、PCR技术检测TRPV6蛋白及m RNA的表达,MTT法、迁移及凋亡实验观察结肠癌SW480细胞增殖、迁移及凋亡变化,高速离子成像系统测定结肠癌SW480细胞内的Ca^(2+)浓度变化。以二甲基肼(dimethyl hydrazine,DMH)建立SD大鼠结肠肿瘤模型,分为实验组(DMH组)和干预组[(DMH+1,25(OH)_2D_3组、DMH+CuCl_2组)]、对照组,干预组分别予1,25(OH)_2D_3(37.5 nmol/kg)、CuCl2(375μmol/kg)对模型进行干预。观察各组大鼠结肠腺瘤及腺癌的发生情况,Western blot检测各组结肠组织中TRPV6蛋白的表达情况。结果:TRPV6-RNAi转染结肠癌SW480细胞后,TRPV6 m RNA及蛋白表达减少,细胞Ca^(2+)浓度水平降低,结肠癌SW480细胞的增殖率、迁移能力下降,细胞凋亡率增加,与空白对照组及阴性对照组比较差异具有统计学意义(P<0.05)。对照组大鼠成瘤率为0,DMH+1,25(OH)_2D_3组为100%,DMH组为84.62%,DMH+CuCl_2组为33.33%。各组大鼠大肠中均有TRPV6蛋白的表达,表达情况为DMH+1,25(OH)_2D_3组>DMH组>DMH+CuCl_2组>对照组,差异具有统计学意义(P<0.05)。结论:降低结肠癌SW480细胞Ca^(2+)浓度水平可抑制结肠癌SW480细胞的增殖、迁移能力,诱导细胞凋亡。1,25(OH)_2D_3可以增加实验鼠结肠组织TRPV6蛋白表达,促进结肠肿瘤的形成。CuCl_2可以使实验鼠结肠组织中的TRPV6蛋白表达降低,并抑制结肠肿瘤的形成。
基金the National Key R&D Program of China(No.2019YFD0900800)the National Science Foundation for Young Scientists of China(No.41606171)+1 种基金the Marine S&T Fund of Shandong Province for Pilot National Laboratory for Marine Science and Technology(Qingdao)(No.2018SDKJ0502)the Science and Technology Service Network Program of Chinese Academy of Sciences(No.KFJ-STS-ZDTP-55)。
文摘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.
