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Influence of CO_2 pneumoperitoneum on intracellular pH and signal transduction in cancer cells 被引量:18
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作者 曹利平 丁国平 +1 位作者 阙日升 郑树 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE EI CAS CSCD 2005年第7期650-655,共6页
Object: The authors studied the influence of CO2 pneumoperitoneum on intracellular pH and signal transduction arising from cancer cell multiplication in laparoscopic tumor operation. Method: They set up a simulation o... Object: The authors studied the influence of CO2 pneumoperitoneum on intracellular pH and signal transduction arising from cancer cell multiplication in laparoscopic tumor operation. Method: They set up a simulation of pneumoperitoneum under different CO2 pressure, and then measured the variation of intracellular pH (pHi) at different time and the activity of protein kinase C (PKC) and protein phosphatase 2a (PP2a) at the end of the pneumoperitoneum. After 1 week, the concentration of cancer cells in the culture medium was calculated. Result: When the pressure of CO2 pneumoperitoneum was 0, 10, 20, 30 mmHg respectively, the average pHi was 7.273, 7.075, 6.783, 6.693 at the end of the pneumoperitoneum; PKC activity was 159.4, 168.5,178.0, 181.6 nmol/(g.min) and PP2a was 4158.3, 4066.9, 3984.0, 3878.5 nmol/(g.min) respectively. After 1 week, the cancer cells concentration was 2.15×105, 2.03×105, 2.20×105, 2.18×105 L-1. Conclusion: CO2 pneumoperitoneum could promote acidosis in cancer cells, inducing the activation of protein kinase C and deactivation of protein phosphatase 2a, but it could not accelerate the mitosis rate of the cancer cells. 展开更多
关键词 caNCER CO2 pneumoperitoneum Intracellular pH signal transduction
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氧糖剥夺再灌注对神经元Wnt5a/FZD2/Ca^(2+)通路的影响
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作者 刘赛赛 张小楠 +4 位作者 李丽 陈洁 赵士弟 黄丽 陶泉坊 《蚌埠医学院学报》 CAS 2024年第5期561-566,共6页
目的:探讨氧糖剥夺再灌注(OGD/Rep)后神经元细胞内Wnt5a/FZD2/Ca^(2+)通路的变化。方法:采用OGD/Rep模型构建神经元细胞局部缺血/再灌注损伤,分正常组、再灌注(0、3、6、9、12、24 h)组。免疫荧光鉴定皮层神经元细胞,CCK-8法确定神经元... 目的:探讨氧糖剥夺再灌注(OGD/Rep)后神经元细胞内Wnt5a/FZD2/Ca^(2+)通路的变化。方法:采用OGD/Rep模型构建神经元细胞局部缺血/再灌注损伤,分正常组、再灌注(0、3、6、9、12、24 h)组。免疫荧光鉴定皮层神经元细胞,CCK-8法确定神经元细胞OGD时间,Western blotting检测非经典Wnt/Ca^(2+)信号通路中Wnt5a、FZD2、IP3-R和p-CaMKⅡ/CaMKⅡ蛋白的变化,免疫荧光和酶标仪检测细胞内Ca^(2+)和活性氧(ROS)的含量。结果:MAP-2免疫荧光鉴定原代皮层神经元细胞纯度较高。与正常组比较,神经元细胞OGD 3 h后细胞存活率为55.46%;与正常组比较,OGD/Rep后非经典Wnt/Ca^(2+)信号通路相关蛋白Wnt5a(3、6、9、12、24 h)、FZD2(3、6、9 h)、IP3-R(6、9、12、24 h)和p-CaMKⅡ/CaMKⅡ(6 h)的相对表达量上调(P<0.05~P<0.01);与正常组比较,OGD/Rep后神经细胞内Ca^(2+)和ROS的表达量均升高(P<0.01)。结论:神经元细胞OGD/Rep能激活Wnt5a/FZD2/Ca^(2+)信号通路。 展开更多
关键词 脑卒中 神经元 氧糖剥夺/复氧 Wnt/ca^(2+)信号通路
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Sericin can reduce hippocampal neuronal apoptosis by activating the Akt signal transduction pathway in a rat model of diabetes mellitus 被引量:4
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作者 Zhihong Chen Yaqiang He +3 位作者 Chengjun Song Zhijun Dong Zhejun Su Jingfeng Xue 《Neural Regeneration Research》 SCIE CAS CSCD 2012年第3期197-201,共5页
In the present study, a rat model of type 2 diabetes mellitus was established by continuous peritoneal injection of streptozotocin. Following intragastric perfusion of sericin for 35 days, blood glucose levels signifi... In the present study, a rat model of type 2 diabetes mellitus was established by continuous peritoneal injection of streptozotocin. Following intragastric perfusion of sericin for 35 days, blood glucose levels significantly reduced, neuronal apoptosis in the hippocampal CA1 region decreased, hippocampal phosphorylated Akt and nuclear factor kappa B expression were enhanced, but Bcl-xL/Bcl-2 associated death promoter expression decreased. Results demonstrated that sericin can reduce hippocampal neuronal apoptosis in a rat model of diabetes mellitus by regulating abnormal changes in the Akt signal transduction pathway. 展开更多
关键词 SERICIN type 2 diabetes mellitus HIPPOcaMPUS apoptosis Akt signal transduction pathway neural regeneration
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CABYR RNAi plasmid construction and NF-κB signal transduction pathway 被引量:2
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作者 Lin-Xiang Shi,Yao-Ming He,Lin Fang,Hong-Bo Meng,Li-Jun Zheng,Department of General Surgery,Shanghai Tenth People’s Hospital,Tongji University,Shanghai 200072,China 《World Journal of Gastroenterology》 SCIE CAS CSCD 2010年第39期4980-4985,共6页
AIM:To construct the CABYR RNAi plasmid and study its relation with the nuclear factor(NF)-κB signal transduction pathway.METHODS:Human CABYR mRNA sequence was obtained from GenBank.The structure of cDNA sequence for... AIM:To construct the CABYR RNAi plasmid and study its relation with the nuclear factor(NF)-κB signal transduction pathway.METHODS:Human CABYR mRNA sequence was obtained from GenBank.The structure of cDNA sequence for the short hairpin RNA was BbsⅠ+sense+loop+ antisense+transcription terminator+KpnⅠ+Bam HⅠ.A CABYR silencing plasmid was constructed and transfected into the human embryo cell line 293T.Quantitative real-time polymerase chain reaction was used to analyze CABYR and NF-κB gene expression.RESULTS:The CABYR and NF-κB expressions were detected in 293T cells.The oligonucleotide(5'-GCT-CAGATGTTAGGTAAAG-3')efficiently silenced the expression of CABYR.The expression of NF-κB was not significantly affected by silencing CABYR(P=0.743).CONCLUSION:CABYR can be found in the human embryo cell line 293T.Cabyrmid 2 can efficiently silence its target,CABYR,indicating that CABYR is not related with the NF-κB signal transduction pathway. 展开更多
关键词 caBYR PLASMID NUCLEAR factor-κB signal transduction RNAI cabyrmid 2
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Effect of MEK1 inhibitor PD98059 on the signal transduction of Tec in HepG2 hepatoma cells 被引量:1
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作者 Jijun Zheng Chuan Chen (Co-First Author) +9 位作者 Ge Wang Jing Deng Jin Yang Hongzhong Wang Minzhi Zhang Qiong Li Wen Xu Qing Hu Zengpeng Li Dong Wang 《The Chinese-German Journal of Clinical Oncology》 CAS 2007年第6期542-545,共4页
Objective: To investigate the effect of MEK1 inhibitor PD98058 on Tec and ERK2 in HepG2 hepatoma cells. Methods: The expression of mRNA and protein of Tec and ERK2 in HepG2 cells was detected by immunocytochemistry as... Objective: To investigate the effect of MEK1 inhibitor PD98058 on Tec and ERK2 in HepG2 hepatoma cells. Methods: The expression of mRNA and protein of Tec and ERK2 in HepG2 cells was detected by immunocytochemistry as- say. After various concentration of PD98059 treatment, the expression of Tec and ERK2 mRNA in HepG2 cells was detected by RT-PCR and Western blotting. Results: Tec and ERK2 expressed highly in HepG2 cells. PD98059 obviously inhibited the expression of mRNA and protein of Tec and ERK2 in a dose-dependent manner, in which 40 μmol/L of PD98059 exhibited the strongest inhibiting effect. Conclusion: PD98058, as MEK1 inhibitor, can inhibit Tec, block the signal route of Ras/Raf/ERK and to impede the signal transduction in HepG2 cells. Tec may be the signal protein in the upper stream of Ras/Raf/ERK in hepatocarcinoma cells and is supposed to interact with the signal way of Ras/Raf/ERK. 展开更多
关键词 PD98059 TEC ERK2 HEPATOcaRCINOMA signal transduction
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基于Ca^(2+)-CaMKⅡ信号通路的济川煎治疗阳虚便秘的作用及分子机制 被引量:3
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作者 刘聪 张悦 +4 位作者 张润涛 荆然 郭丁丁 倪艳 康永 《中国药理学通报》 CAS CSCD 北大核心 2023年第2期366-372,共7页
目的基于Ca^(2+)-CaMKⅡ通路观察济川煎对阳虚便秘大鼠的治疗作用及分子机制。方法采用白醋+活性炭冰水联合复方地芬诺酯片复制阳虚便秘大鼠模型。将模型成功大鼠随机分为5组,即模型组、莫沙必利组(1.88 mg·kg^(-1))、济川煎高(8.2... 目的基于Ca^(2+)-CaMKⅡ通路观察济川煎对阳虚便秘大鼠的治疗作用及分子机制。方法采用白醋+活性炭冰水联合复方地芬诺酯片复制阳虚便秘大鼠模型。将模型成功大鼠随机分为5组,即模型组、莫沙必利组(1.88 mg·kg^(-1))、济川煎高(8.26 g·kg^(-1))、中(4.31 g·kg^(-1))、低(2.16 g·kg^(-1))剂量组,灌胃给药每日1次,连续7 d。末次给药,记录大鼠状态并评分,进行排便功能测定;分离模型大鼠结肠平滑肌细胞并鉴定,FCM检测结肠平滑肌细胞内Ca^(2+)浓度变化;采用ELISA法检测大鼠血清中cAMP、cGMP含量及其比值;采用RT-PCR及Western blot法检测大鼠结肠平滑肌细胞CaM、CaMKⅡ亚型的表达情况。结果济川煎可明显缓解模型大鼠腹胀,摄食、饮水量、排便减少及体质量降低的症状(P<0.05),评分明显升高(P<0.01);能明显缩短首粒排便时间(P<0.01),增加排便粒数(P<0.05);济川煎还可明显增加模型大鼠血清中cAMP含量(P<0.05),减少cGMP含量(P<0.01),cAMP/cGMP比值明显增加(P<0.01);明显降低大鼠结肠平滑肌细胞内Ca^(2+)浓度(P<0.01);PCR及Western blot检测结果显示,济川煎可明显降低CaM及CaMKⅡβ、γ、δ亚型蛋白表达(P<0.01)。结论济川煎对阳虚便秘具有明显的治疗作用,可能与其调节Ca^(2+)-CaMKⅡ信号通路有关。 展开更多
关键词 济川煎 阳虚便秘 ca^(2+)-caMKⅡ 治疗作用 信号通路 分子机制
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苜蓿体内H_(2)S信号与Ca^(2+)调节气孔运动的作用机制 被引量:1
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作者 郝雪峰 亢春霞 +1 位作者 裴雁曦 金竹萍 《植物研究》 CAS CSCD 北大核心 2023年第2期281-287,共7页
为探究H_(2)S信号在苜蓿(Medicago sativa)体内调节气孔运动的作用,及在此过程中H_(2)S与Ca^(2+)的关系,以蒺藜苜蓿(Medicago truncatula)的野生型和钙离子转运体突变体为试验材料,分别从转录水平、细胞水平和生理水平开展研究。采用qRT... 为探究H_(2)S信号在苜蓿(Medicago sativa)体内调节气孔运动的作用,及在此过程中H_(2)S与Ca^(2+)的关系,以蒺藜苜蓿(Medicago truncatula)的野生型和钙离子转运体突变体为试验材料,分别从转录水平、细胞水平和生理水平开展研究。采用qRT-PCR比较相关基因的表达量变化、荧光探针显示体内Ca^(2+)含量、电极法测定H_(2)S含量、光学显微镜观察和测量气孔孔径等。结果表明:蒺藜苜蓿突变体NF3011和NF2734体内H_(2)S的含量与野生型相比极显著降低(P<0.01);H_(2)S信号在一定程度上抑制钙离子转运体编码基因MTR_6g027580的表达;外源生理浓度H_(2)S熏蒸可诱导蒺藜苜蓿气孔关闭,与Ca^(2+)通道阻断剂LaCl_(3)联合处理对野生型气孔运动未产生影响,而在突变体中的结果截然相反;利用荧光探针测定保卫细胞内的Ca^(2+)含量,所得结果与气孔孔径的变化规律完全一致。综上所述,H_(2)S信号促进叶片保卫细胞内Ca^(2+)的含量增加,最终表现为植物气孔孔径变小,在此过程中胞内Ca^(2+)含量变化主要通过Ca^(2+)转运体进行,少部分依赖Ca^(2+)离子通道。该研究结果不仅在理论上丰富了H_(2)S信号的作用机制,更具应用于苜蓿生产实践并推广于其他作物的潜力。 展开更多
关键词 H_(2)S信号 ca^(2+) 气孔运动 蒺藜苜蓿
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基于Hepcidin和JAK2/STAT3信号通路探讨通痹颗粒 对胶原诱导性关节炎大鼠的影响
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作者 吴伊莹 柳玉佳 +2 位作者 廖亮英 范伏元 郭志华 《湖南中医药大学学报》 CAS 2024年第6期960-966,共7页
目的研究通痹颗粒对胶原诱导性关节炎(collagen-induced arthritis,CIA)大鼠铁调素(hepcidin,Hepc)、Janus激酶(janus kinase,JAK)2/信号转导子和转录激活子(signal transduction and activator of transcription,STAT)3信号通路的影响... 目的研究通痹颗粒对胶原诱导性关节炎(collagen-induced arthritis,CIA)大鼠铁调素(hepcidin,Hepc)、Janus激酶(janus kinase,JAK)2/信号转导子和转录激活子(signal transduction and activator of transcription,STAT)3信号通路的影响。方法选取36只雌性SD大鼠随机分成空白组、模型组、阳性对照组和通痹颗粒低、中、高剂量组,每组6只。空白组不予处理,其余组用牛Ⅱ型胶原建立CIA模型。造模完成后,空白组、模型组予生理盐水灌胃,其余各组分别以巴瑞替尼片和低、中、高剂量通痹颗粒灌胃。每天1次,连续4周。HE染色行滑膜组织病理学观察;酶联免疫吸附法测定血清Hepc、白细胞介素6(interleukin 6,IL-6)水平;逆转录-聚合酶链反应法测定滑膜中JAK2、STAT3、细胞信号因子传导抑制体(suppressor of cytokine signaling,SOCS)1、SOCS3的mRNA相对表达量;Western blot法检测滑膜中JAK2、p-JAK2、STAT3、p-STAT3、SOCS1、SOCS3的蛋白表达量。结果模型组见滑膜上皮结构缺损,滑膜重度增生,排列紊乱,并有大量炎症细胞浸润和多个血管翳形成;各给药组滑膜炎症均有所减轻,阳性对照组优于通痹颗粒高剂量组,通痹颗粒中、高剂量组优于低剂量组。与模型组相比,各给药组关节炎指数评分、血清Hepc和IL-6水平均显著降低(P<0.01);与阳性对照组相比,通痹颗粒中、低剂量组关节炎指数评分、血清Hepc和IL-6水平均升高(P<0.05)。与模型组比较,阳性对照组和通痹颗粒低、中、高剂量组JAK2、STAT3 mRNA和蛋白以及p-JAK2、p-STAT3的蛋白表达量均降低(P<0.05),而通路抑制因子SOCS1、SOCS3 mRNA和蛋白的表达均升高(P<0.05);与阳性对照组比较,通痹颗粒各剂量组JAK2、STAT3 mRNA和蛋白以及p-JAK2、p-STAT3的蛋白表达量均升高(P<0.05),而SOCS1、SOCS3 mRNA和蛋白的表达均降低(P<0.05)。结论通痹颗粒能够改善CIA大鼠滑膜炎症,其机制可能与抑制JAK2/STAT3信号通路而减少Hepc的表达有关。 展开更多
关键词 类风湿关节炎 胶原诱导性关节炎 中药 通痹颗粒 铁调素 JAK2/STAT3信号通路
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膀胱癌100例血清Zeste基因增强子同源物2和信号传导蛋白3表达水平及诊断价值分析
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作者 吴娟 王丹 +1 位作者 张前进 裴兵 《安徽医药》 CAS 2024年第10期2035-2038,共4页
目的探讨膀胱癌病人血清Zeste基因增强子同源物2(EZH2)和信号传导蛋白3(SMAD3)表达水平及临床意义。方法纳入江苏省人民医院宿迁医院于2021年1月至2022年12月收治的膀胱癌病人进行研究(100例),另选取同期于该院就诊的泌尿系统良性疾病... 目的探讨膀胱癌病人血清Zeste基因增强子同源物2(EZH2)和信号传导蛋白3(SMAD3)表达水平及临床意义。方法纳入江苏省人民医院宿迁医院于2021年1月至2022年12月收治的膀胱癌病人进行研究(100例),另选取同期于该院就诊的泌尿系统良性疾病病人作为良性疾病组(93例)以及健康体检者作为对照(100例)。采用Pearson相关性分析法分析膀胱癌病人血清中EZH2和SMAD3表达水平的相关性;采用logistic多因素回归分析法分析膀胱癌发生的影响因素;采用ROC曲线分析EZH2和SMAD3对膀胱癌的诊断效能。结果膀胱癌病人血清中EZH2(101.34±15.09)ng/L和SMAD3(226.53±25.94)ng/L表达水平均高于良性疾病组(85.96±11.23)ng/L、(203.11±22.18)ng/L和对照组(83.91±9.14)ng/L、(198.03±20.30)ng/L(均P<0.001);膀胱癌病人血清EZH2和SMAD3表达水平呈正相关(r=0.72,P<0.001);膀胱癌组年龄≥60岁(74/187)、有吸烟史的病人(76/166)所占比例均高于良性疾病组(65/187、55/166)及对照组(48/187、35/166)(均P<0.05);年龄、吸烟史、EZH2、SMAD3为发生膀胱癌的危险因素(P<0.05);以良性疾病组为对照,血清EZH2和SMAD3单独检测诊断膀胱癌的曲线下面积(AUC)分别为0.78、0.78,二者联合检测的AUC为0.85,优于各自单独检测(Z_(二者联合-EZH2)=2.81、Z_(二者联合-SMAD3)=2.68,P=0.009、0.007)。结论血清EZH2和SMAD3与膀胱癌的发生有关,二者联合对膀胱癌具有一定的诊断价值。 展开更多
关键词 膀胱肿瘤 Zeste基因增强子同源物2 信号传导蛋白3 诊断价值 表达水平
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CytochalasinH对小鼠脾细胞增殖的抑制作用及其对Ca^(2+)/CaN/NFAT信号通路的影响 被引量:1
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作者 邓小林 饶乐怡 徐静 《广东海洋大学学报》 CAS CSCD 北大核心 2023年第4期19-26,共8页
【目的】研究红树林植物大红树内生真菌Phomopsis asparagi DHS-48得到的次级代谢产物cytochalasin H对小鼠(Mus musculus)脾细胞的免疫抑制活性及其作用机制。【方法】用不同浓度(3、6、12、25、30μmol/L)cytochalasin H处理小鼠脾细... 【目的】研究红树林植物大红树内生真菌Phomopsis asparagi DHS-48得到的次级代谢产物cytochalasin H对小鼠(Mus musculus)脾细胞的免疫抑制活性及其作用机制。【方法】用不同浓度(3、6、12、25、30μmol/L)cytochalasin H处理小鼠脾细胞,通过CCK-8活力测定法检测其对刀豆蛋白A(Concanavalin A,ConA)诱导小鼠脾细胞增殖的抑制作用;流式细胞术检测cytochalasin H对小鼠脾细胞凋亡的影响;免疫荧光实验检测cytochalasin H对细胞中NFAT入核及钙离子内流的影响;酶联免疫吸附法(Enzyme-linked immune sorbent assay,ELISA)和免疫蛋白印迹法(Western blotting)测定cytochalasin H对脾细胞中NFAT信号通路的影响。【结果】CCK-8实验表明,cytochalasin H对于正常小鼠脾淋巴细胞的毒性比环孢菌素(Cyclosporin A,Cs A)弱,半抑制浓度IC_(50)=(35.07±1.16)μmol/L;流式细胞术显示,cytochalasin H对ConA刺激的脾淋巴细胞无促凋亡作用;cytochalasin H对ConA诱导增殖的小鼠脾淋巴细胞具有显著的抑制效果,IC_(50)=(14.81±0.81)μmol/L,且呈剂量依赖性;免疫荧光结果表明,cytochalasin H在浓度为15μmol/L时显著地抑制了钙离子内流;Western Blotting结果显示cytochalasin H对脾淋巴细胞中CaN、NFAT蛋白(nuclear factor of activated T-cells,NFAT)的表达具有显著的抑制作用;免疫荧光结果显示,cytochalasin H抑制细胞浆内的NFAT转移到细胞核;ELISA测试结果证明,cytochalasin H能显著抑制下游细胞因子白介素-2(IL-2)的分泌(α=0.05)。【结论】Cytochalasin H通过抑制Ca^(2+)/CaN/NFAT信号通路转导显著抑制T细胞增殖和活化。 展开更多
关键词 cytochalasinH T细胞增殖 细胞凋亡 ca^(2+)/caN/NFAT信号通路 免疫抑制剂
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Overexpression of MdFRK2 enhances apple drought resistance by promoting carbohydrate metabolism and root growth under drought stress 被引量:1
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作者 Jingjing Yang Ruiling Zhan +4 位作者 Lan Wang Junqiao Li Baiquan Ma Fengwang Ma Mingjun Li 《Horticultural Plant Journal》 SCIE CAS CSCD 2023年第5期884-897,共14页
Soluble sugars function not only as the energy and structural blocks supporting plants,but also as osmoregulators and signal molecules during plant adaptation to water deficit.Here,we investigated drought resistance i... Soluble sugars function not only as the energy and structural blocks supporting plants,but also as osmoregulators and signal molecules during plant adaptation to water deficit.Here,we investigated drought resistance in transgenic apple(Malus×domestica)overexpressing MdFRK2,a key gene regulating fructose content and sugar metabolism.There is no obvious phenotypic difference between MdFRK2-overexpressing transgenic plants and WT plants under the well-watered condition.However,the transgenic plants and the grafted plants using MdFRK2-overexpressing rootstock exhibited improved tolerance to drought stress.Overexpression of MdFRK2 significantly promoted the growth of root system under drought stress.RNA sequencing showed that under drought stress,genes involved in sugar metabolism,transcription regulation,signal transduction or hormone metabolism were differentially expressed in MdFRK2 transgenic plants.Consistent with the gene expression profile,the activities of enzyme(SDH,FRK and NI)involved in sugar metabolism in the roots of MdFRK2 transgenic plants were significantly higher than those of untransformed control plants after drought stress.Under drought stress,overexpression of MdFRK2 promoted the accumulation of IAA,and decreased the contents of ABA and CK in apple root system.In conclusion,these results suggest that MdFRK2 can promote the growth of apple roots under drought stress by regulating sugar metabolism and accumulation,hormone metabolism and signal transduction,and then resist drought stress. 展开更多
关键词 Malus×domestica MdFRK2 Drought stress Sugar AUXIN signal transduction TRANSCRIPTOME
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肠胶质细胞系TRPV4依赖性Ca^(2+)内流、ATP释放与炎症因子表达 被引量:2
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作者 谭千山 陈君 +4 位作者 谢会超 李立奇 陈帅帅 董辉 肖卫东 《陆军军医大学学报》 CAS CSCD 北大核心 2023年第5期417-425,共9页
目的探讨瞬时感受器电位离子通道香草素受体4(transient receptor potential vanilloid 4,TRPV4)在调节肠胶质细胞外Ca^(2+)内流中的作用及其意义。方法收集大鼠肠胶质细胞系CRL-2690、大鼠肠上皮细胞系IEC6和人胚肾细胞系HEK293T样本,... 目的探讨瞬时感受器电位离子通道香草素受体4(transient receptor potential vanilloid 4,TRPV4)在调节肠胶质细胞外Ca^(2+)内流中的作用及其意义。方法收集大鼠肠胶质细胞系CRL-2690、大鼠肠上皮细胞系IEC6和人胚肾细胞系HEK293T样本,采用RT-qPCR、Western blot和免疫荧光方法检测TRPV4的mRNA、蛋白表达与定位;将CRL-2690细胞依次分为GSK组与GSK+HC组,低渗组与低渗+HC组,GdCl_(3)组与GdCl_(3)+HC组以及CaCl_(2)组与CaCl^(2+)HC组,采用Fura-2荧光探针和单细胞荧光Ca^(2+)检测仪评估CRL-2690细胞中TRPV4通道活性。将CRL-2690细胞分为对照组、GSK组与GSK+HC组,通过荧光素-荧光素酶实验测量细胞ATP的释放水平;采用RT-qPCR检测炎症相关基因GFAP、IL-1β、IL-6、IL-10的mRNA表达。结果在大鼠肠胶质细胞系CRL-2690中检测到TRPV4 mRNA和蛋白表达;TRPV4特异性激动剂GSK1016790A和低渗透压刺激明显增强了CRL-2690细胞内钙荧光强度,而TRPV4特异性抑制剂HC067047可抑制此作用(0.43038±0.06365 vs 0.00423±0.00578,P<0.0001);钙敏感受体(calcium-sensing receptor,CaSR)激活诱导的外Ca^(2+)内流也可被HC067047所抑制(P<0.05);功能上,激活TRPV4可促ATP释放、EGC反应性增生标志物GFAP和炎症因子IL-1β、IL-6的表达,并抑制抗炎因子IL-10的表达,该作用可被HC067047所抑制(P<0.05)。结论激活TRPV4可引起肠胶质细胞外Ca^(2+)内流,并促进ATP释放与炎症因子表达,可能参与肠道炎症的发生。 展开更多
关键词 肠胶质细胞 瞬时感受器电位离子通道香草素受体4 ca^(2+)信号 三磷酸腺苷 炎症
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BDNF经TrkB-PKC-Ca^(2+)信号通路调控气道ASMC炎症反应
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作者 聂颖 徐辉甫 +3 位作者 王丹 张领领 邵会 乔静(指导) 《中国免疫学杂志》 CAS CSCD 北大核心 2023年第3期531-536,共6页
目的:探讨脑源性神经营养因子(BDNF)经TrkB-PKC-Ca^(2+)信号影响气道细胞收缩及炎症反应的分子机制。方法:建立TNF-α诱导的气道平滑肌细胞(ASMC)炎症模型,采用BDNF-siRNA和BDNF重组蛋白对模型进行干预,测定各组ASMC炎症因子IL-17、Eota... 目的:探讨脑源性神经营养因子(BDNF)经TrkB-PKC-Ca^(2+)信号影响气道细胞收缩及炎症反应的分子机制。方法:建立TNF-α诱导的气道平滑肌细胞(ASMC)炎症模型,采用BDNF-siRNA和BDNF重组蛋白对模型进行干预,测定各组ASMC炎症因子IL-17、Eotaxin水平及BDNF、TrkB浓度、PKC活性变化及Ca^(2+)、三磷酸肌醇(IP3)浓度变化。结果:与正常组相比,模型组细胞异常增殖,细胞内炎症因子IL-17、Eotaxin增加,BDNF、TrkB蛋白和转录水平明显升高,同时引起其下游PKCIP3-Ca^(2+)信号上调。结论:BDNF可能通过激活下游PKC信号通路诱发气道重塑,参与气道高反应形成。 展开更多
关键词 BDNF TrkB-PKC-ca^(2+)信号 ASMC 气道炎症反应
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The SHP-2 tyrosine phosphatase:Signaling mechanisms and biological functions 被引量:25
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作者 Cheng Kui QU 《Cell Research》 SCIE CAS CSCD 2000年第4期279-288,共10页
Cellular biological activities are tightly controlled by intracellular signaling processes initiated by extracellular signals. Protein tyrosine phosphatases, which remove phosphate groups from phosphorylated signaling... Cellular biological activities are tightly controlled by intracellular signaling processes initiated by extracellular signals. Protein tyrosine phosphatases, which remove phosphate groups from phosphorylated signaling molecules, play equally important tyrosine roles as protein tyrosine kinases in signal transduction. SHP-2, a cytoplajsmic SH2 domain containing protein tyrosine phosphatase, is involved in the signaling pathways of a variety of growth factors and cytokines. Recent studies have clearly demonstrated that this phosphatase plays an important role in transducing signal relay from the cell surface to the nucleus, and is a critical intracellular regulator in mediating cell proliferation and differentiation. 展开更多
关键词 SHP-2 SHP-1 signal transduction
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Primary evidence for involvement of IP_(3) in heat-shock signal transduction in Arabidopsis 被引量:9
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作者 Hong Tao Liu Fei Gao +3 位作者 Shu Juan Cui Jin Long Han Da Ye Sun Ren Gang Zhou 《Cell Research》 SCIE CAS CSCD 2006年第4期394-400,共7页
The role of inositol 1,4,5-trisphosphate (IP3) in transducing heat-shock (HS) signals was examined in Arabidopsis. The whole-plant IP3 level increased within 1 min of HS at 37℃. After 3 min of HS, the IP3 level r... The role of inositol 1,4,5-trisphosphate (IP3) in transducing heat-shock (HS) signals was examined in Arabidopsis. The whole-plant IP3 level increased within 1 min of HS at 37℃. After 3 min of HS, the IP3 level reached a maximum 2.5 fold increase. Using the transgenic Arabidopsis plants that have AtHsp 18.2 promoter-β-glucuronidase (GUS) fusion gene, it was found that the level of GUS activity was up-regulated by the addition of caged IP3 at both non-HS and HS temperatures and was down-regulated by the phospholipase C (PLC) inhibitors {1-[6-(( 1713-3-Methoxyestra-1,3,5(10)-trien- 7-yl)amino)hexyl]-2,5-pyrrolidinedione } (U-73122). The intracellular-free calcium ion concentration ([Ca^2+]i) increased during HS at 37℃ in suspension-cultured Arabidopsis cells expressing apoaequorin. Treatment with U-73122 prevented the increase of [Ca^2+]i to some extent. Above results provided primary evidence for the possible involvement of IP3 in HS signal transduction in higher plants. 展开更多
关键词 heat shock signal transduction IP3 PLC [ca^2+]i ARABIDOPSIS
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Ca^(2+) involvement in activation of extracellular-signal- regulated-kinase 1/2 and m-calpain after axotomy of the sciatic nerve 被引量:4
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作者 Lisa B. Martensson Charlotta Lindwall Blom Lars B. Dahlin 《Neural Regeneration Research》 SCIE CAS CSCD 2017年第4期623-628,共6页
Detailed mechanisms behind regeneration after nerve injury, in particular signal transduction and the fate of Schwann cells (SCs), are poorly understood. Here, we investigated axotomy-induced activation of extracell... Detailed mechanisms behind regeneration after nerve injury, in particular signal transduction and the fate of Schwann cells (SCs), are poorly understood. Here, we investigated axotomy-induced activation of extracellular- signal-regulated kinase-1/2 (ERK1/2; important for proliferation) and m-calpain in vitro, and the relation to Ca2+ deletion and Schwann cell proliferation and death after rat sciatic nerve axotomy. Nerve segments were cultured for up to 72 hours with and without ethylene glycol-bis(β-aminoethyl ether)- N,N,N',N'-tetraacetic acid (EGTA). In some experiments, 5-bromo-2′-deoxyuridine (BrdU) was added during the last 24 hours to detect proliferating cells and propidium iodide (PI) was added at the last hour to detect dead and/or dying cells. Immunohistochemistry of sections of the cultured nerve segments was performed to label m-calpain and the phosphorylated and activated form of ERK1/2. The experiments revealed that immunoreactivity for p-ERK1/2 increased with time in organotypically cultured SCs. p-ERK1/2 and m-calpain were also observed in axons. A significant increase in the number of dead or dying SCs was observed in nerve segments cultured for 24 hours. When deprived of Ca2+, activation of axonal m-calpain was reduced, whereas p-ERK1/2 was increased in SCs. Ca2+ deprivation also significantly reduced the number of proliferating SCs, and instead increased the number of dead or dying SCs. Ca2+ seems to play an important role in activation of ERK1/2 in SCs and in SC survival and proliferation. In addition, extracellular Ca2+ levels are also required for m-calpain activation and up-regulation in axons. Thus, regulation of Ca2+ levels is likely to be a useful method to promote SC proliferation. 展开更多
关键词 nerve regeneration P-ERK1/2 M-caLPAIN nerve injury signal transduction cell proliferation cell death ACTIVATION AXOTOMY sciatic nerve neural regeneration
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Leptin transiently antagonizes ghrelin and long-lastingly orexin in regulation of Ca^(2+) signaling in neuropeptide Y neurons of the arcuate nucleus 被引量:4
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作者 Daisuke Kohno Shigetomo Suyama Toshihiko Yada 《World Journal of Gastroenterology》 SCIE CAS CSCD 2008年第41期6347-6354,共8页
AIM: To explore the mechanism for interactions of leptin with ghrelin and orexin in the arcuate nucleus (ARC) activating neuropeptide Y (NPY) neurons during physiological regulation of feeding, METHODS: Single n... AIM: To explore the mechanism for interactions of leptin with ghrelin and orexin in the arcuate nucleus (ARC) activating neuropeptide Y (NPY) neurons during physiological regulation of feeding, METHODS: Single neurons from ARC of adult rats with matured feeding function were isolated. [Ca2+]i was measured to monitore their activities. The time course of leptin effects on ghrelin-induced versus orexin-induced [Ca2+]i increases in NPY neurons was studied. RESULTS: Administration of ghrelin or orexin-A at 101~ mol/L increased cytosolic Ca2~ concentration ([Ca2+]~) in NPY neurons isolated from the ARC of adult rats. Upon administration of leptin at 10^-14-10^-12 mol/L, ghrelin-induced [Ca2+]i increases were initially (〈 10 min) inhibited but later restored, exhibiting a transient pattern of inhibition. In contrast, orexin-induced [Ca2+]i increases were inhibited by leptin in a long- lasting manner. Furthermore, a prior administration of leptin inhibited orexin action but not ghrelin action to increase ICa 2+li, CONCLUSION: Leptin counteracted ghrelin effects transiently and orexin effects long-lastingly in NPY neurons. The transient property with which leptin counteracts ghrelin action in NPY neurons may allow the fasting-associated increase in ghrelin levels to activate NPY neurons in the presence of physiological leptin and to stimulate feeding. 展开更多
关键词 LEPTIN GHRELIN OREXIN Arcuate nucleus Neuropeptide Y ca2+ FEEDING Phosphatidylinositol 3-kinase Phosphodiesterase 3 signal transducer and activator of transcription 3
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Update on vascular endothelial Ca^(2+) signalling:A tale of ion channels,pumps and transporters 被引量:2
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作者 Francesco Moccia Roberto Berra-Romani Franco Tanzi 《World Journal of Biological Chemistry》 CAS 2012年第7期127-158,共32页
A monolayer of endothelial cells (ECs) lines the lumen of blood vessels and forms a multifunctional transducing organ that mediates a plethora of cardiovascular processes. The activation of ECs from as state of quiesc... A monolayer of endothelial cells (ECs) lines the lumen of blood vessels and forms a multifunctional transducing organ that mediates a plethora of cardiovascular processes. The activation of ECs from as state of quiescence is, therefore, regarded among the early events leading to the onset and progression of potentially lethal diseases, such as hypertension, myocardial infarction, brain stroke, and tumor. Intracellular Ca2+ signals have long been know to play a central role in the complex network of signaling pathways regulating the endothelial functions. Notably, recent work has outlined how any change in the pattern of expression of endothelial channels, transporters and pumps involved in the modulation of intracellular Ca2+ levels may dramatically affect whole body homeostasis. Vascular ECs may react to both mechanical and chemical stimuli by generating a variety of intracellular Ca2+ signals, ranging from brief, localized Ca2+ pulses to prolonged Ca2+ oscillations engulfing the whole cytoplasm. The well-defined spatiotemporal profile of the subcellular Ca2+ signals elicited in ECs by specific extracellular inputs depends on the interaction between Ca2+ releasing channels, which arelocated both on the plasma membrane and in a number of intracellular organelles, and Ca2+ removing systems. The present article aims to summarize both the past and recent literature in the field to provide a clear-cut picture of our current knowledge on the molecular nature and the role played by the components of the Ca2+ machinery in vascular ECs under both physiological and pathological conditions. 展开更多
关键词 Endothelial cells ca2+ signalLING Plasma membrane Endoplasmic reticulum Intracellular ca2+ release ca2+ entry ca2+ removal ca2+ oscillations
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基于JAK2/STAT3通路探讨阿奇霉素对脂多糖诱导肺泡上皮细胞的保护作用
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作者 温玲 李宝琪 +2 位作者 赵艳敏 郑舒扬 苏颖 《四川医学》 CAS 2024年第3期263-268,共6页
目的探究阿奇霉素(AZI)对脂多糖(LPS)诱导的人肺泡上皮细胞增殖生长、迁移的作用及相关机制。方法体外培养人肺泡上皮细胞A549,分为对照组(不做干预)、LPS组(10μg/ml LPS处理24 h)、低/中/高剂量实验组(10μg/ml LPS+2、4、8μg/ml AZI... 目的探究阿奇霉素(AZI)对脂多糖(LPS)诱导的人肺泡上皮细胞增殖生长、迁移的作用及相关机制。方法体外培养人肺泡上皮细胞A549,分为对照组(不做干预)、LPS组(10μg/ml LPS处理24 h)、低/中/高剂量实验组(10μg/ml LPS+2、4、8μg/ml AZI)、AZI组(10μg/ml LPS+4μg/ml AZI)、抑制剂组(10μg/ml LPS+50μmol/L JAK2/STAT3通路抑制剂AG490)、AZI+抑制剂组(10μg/ml LPS+4μg/ml AZI+50μmol/L AG490)、AZI+激活剂组(10μg/ml LPS+4μg/ml AZI+0.5μmol/L JAK2/STAT3通路激活剂Colivelin)。干预24 h后,采用细胞计数试剂盒-8(CCK-8)、酶联免疫吸附试验(ELISA)、倒置显微镜、划痕法、蛋白免疫印迹(WB)法检测炎症因子肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β和IL-6表达水平、细胞生长、迁移率、上皮间质转化(EMT)及JAK2/STAT3信号通路相关蛋白表达水平。结果LPS组细胞活力较对照组下降(P<0.05)。中/高剂量实验组细胞活力较LPS组上升(P<0.05)。因此选择有显著差异的较低浓度(4μg/ml AZI)作为AZI组进行后续实验。与对照组相比,LPS组细胞生长受抑制,TNF-α、IL-1β、IL-6、E-钙黏蛋白(E-cadherin)表达降低(P<0.05),细胞迁移率、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、纤维粘连蛋白(FN)、p-JAK2、p-STAT3蛋白表达升高(P<0.05)。与LPS组相比,AZI组和抑制剂组显著扭转了上述指标的变化(P<0.05)。与AZI组相比,AZI+抑制剂组细胞生长状态较好,E-cadherin蛋白表达进一步升高(P<0.05),细胞迁移率、TNF-α、IL-1β、IL-6、N-cadherin、Vimentin、FN、p-JAK2、p-STAT3蛋白表达进一步降低(P<0.05),AZI+激活剂组则显著逆转了上述指标的变化(P<0.05)。结论阿奇霉素能够通过抑制JAK2/STAT3信号通路减轻对A549细胞的炎症损伤,促进细胞生长,并抑制其迁移与上皮间质转化(EMT)进程。 展开更多
关键词 阿奇霉素 肺泡上皮细胞 Janus激酶2/信号转导和转录启动因子3 迁移 上皮间质转化
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基于生物信息学分析探讨羟基红花黄色素A通过JAK2/STAT3信号通路抑制缺血性脑卒中后神经元凋亡的机制
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作者 吴艺舸 殷丽君 +4 位作者 王泽乾 贾思锋 温春丽 宋丽娟 马存根 《中西医结合心脑血管病杂志》 2024年第18期3326-3335,3352,共11页
目的:利用生物信息学技术筛选并鉴定缺血性脑卒中(IS)的关键基因,基于基因富集分析结合相关实验探讨羟基红花黄色素A(HSYA)对脑缺血损伤后神经元凋亡的影响及机制。方法:从GEO数据库获得IS相关的样本数据,进行差异表达基因(DEGs)分析及... 目的:利用生物信息学技术筛选并鉴定缺血性脑卒中(IS)的关键基因,基于基因富集分析结合相关实验探讨羟基红花黄色素A(HSYA)对脑缺血损伤后神经元凋亡的影响及机制。方法:从GEO数据库获得IS相关的样本数据,进行差异表达基因(DEGs)分析及基因富集分析,获得关键基因与关键信号通路,并通过体内外实验进行相关验证。建立Sprague-Dawley大鼠大脑中动脉闭塞再灌注模型(MCAO/R),采用蛋白免疫印迹法(Western Blot)和免疫荧光检测Janus激酶2(JAK2)/信号传导转录激活因子3(STAT3)通路的磷酸化水平及凋亡相关蛋白的表达;线粒体膜电位探针检测线粒体膜电位情况进而明确细胞凋亡水平。利用HT-22海马神经元细胞建立糖氧剥夺/复糖氧模型(OGD/R),使用JAK2/STAT3信号通路抑制剂进一步验证HSYA对神经元凋亡的作用机制。结果:通过生物信息学分析研究GSE22255数据集发现23个显著上调的DEGs和2个显著下调的DEGs。富集分析发现其与脂多糖的反应、细胞凋亡的调控、炎症反应、急性炎症反应调节、分子干预信号通路相关。生物过程方面,通过建立特征基因功能网络发现,特征基因与急性炎症反应、凋亡信号通路的调节、脂多糖介导的反应、稳态分子的反应等功能相关。基因集富集分析显示,肿瘤坏死因子α(TNF-α)介导的核因子κB(NF-κB)信号通路、血红素代谢信号通路、细胞凋亡相关信号通路、JAK/STAT3信号通路、P53信号通路发挥着关键作用。筛选出JAK2/STAT3信号通路和凋亡相关通路为研究重点。与假手术组比较,MCAO/R组大鼠磷酸化JAK2(p-JAK2)、磷酸化STAT3(p-STAT3)和促凋亡相关蛋白表达升高(P<0.001),抑凋亡相关蛋白表达降低(P<0.001)。HSYA干预后可抑制JAK2/STAT3信号通路磷酸化激活和神经元凋亡(P<0.01)。体外实验显示,OGD/R组JAK2/STAT3通路被磷酸化激活,促凋亡相关蛋白表达较Normal组升高(P<0.001),抑凋亡相关蛋白表达低于Normal组(P<0.001);加入抑制剂AG490后JAK2、STAT3磷酸化程度降低(P<0.01)。与Normal组比较,OGD/R组凋亡相关蛋白半胱氨酸蛋白酶3(Cleaved Caspase-3)、Bcl-2关联X蛋白(Bax)表达水平升高(P<0.001),Bcl-2表达降低(P<0.001)。HSYA抑制了神经元的凋亡(P<0.01)。结论:JAK2/STAT3信号通路和凋亡相关信号通路在IS后发挥着关键作用,HSYA可能通过调控JAK2/STAT3信号通路,抑制缺血缺氧后神经元凋亡,从而减轻脑损伤。 展开更多
关键词 缺血性脑卒中 生物信息学分析 羟基红花黄色素A 脑缺血 神经元 Janus激酶2/信号传导转录激活因子3信号通路 凋亡 实验研究
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