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作 者:杨雯雨 马新润 朱勇[1] 孟潇潇[1] 杨正峰 田锐[1] YANG Wen-yu;MA Xin-run;ZHU Yong;MENG Xiao-xiao;YANG Zheng-feng;TIAN Rui(Department of Crutical Care Medicine,Shanghai General Hospital,Shanghai Jiaotong University,School of Medicine,Shanghai,201620,China;Institute of Clinical Immunology,Center for Translational Medicine,Shanghai General Hospital,Shanghai Jiaotong University,School of Medicine,Shanghai,201620,China)
机构地区:[1]上海交通大学附属第一人民医院急诊危重病科,上海201620 [2]上海交通大学附属第一人民医院临床转化研究院,上海201620
出 处:《现代生物医学进展》2021年第21期4006-4010,共5页Progress in Modern Biomedicine
基 金:国家自然科学基金项目(81971555)。
摘 要:目的:探讨SOCE对百草枯(PQ)致肺纤维化过程的具体调控作用。方法:体外培养肺泡上皮A549细胞,设置分组为对照组、PQ组和PQ+SKF96365组。对照组不作任何处理;PQ组给予800μM的PQ溶液处理细胞24 h;PQ+SKF96365组先加入10μM的SKF96365预处理细胞2 h,然后再加800μM PQ处理24 h。免疫荧光检测PQ染毒后SOCE相关蛋白的活化和胞内分布、NFATc1的表达和入核情况。清洁级小鼠40只,随机分为空白对照组、PQ组、SKF96365组和PQ+SKF96365组。PQ组及PQ+SKF96365组给予百草枯(20 mg/kg)一次性腹腔注射,对照组给予等量生理盐水;SKFS96365组及PQ+SKF96365组每天腹腔注射一次10 mg/kg的SKF96365溶液,持续3天,行HE染色、Masson染色观察肺组织病理状态改变以及胶原纤维的变化情况。结果:与正常组相比,PQ染毒组STIM1蛋白活化出现寡聚化现象并且ORAI1、TRPC1膜定位显著;PQ引起细胞质内NFATc1信号向细胞核内转移,密度差异具有统计学意义(P<0.01),使用SOCE抑制剂后NFATc1信号减弱向细胞质内转移(P<0.01)。与对照组相比,PQ中毒的小鼠肺泡结构被破坏,肺间质大量胶原纤维沉积;与PQ组相比,PQ+SKF96365组肺泡结构相对完整并且间质胶原纤维沉积减少。结论:PQ中毒可活化SOCE并通过增加NFAT入核激活下游转录信号,促进肺纤维化的发生。Objective:To explore the effect of SOCE in PQ-induced pulmonary fibrosis.Methods:A549 cells were cultured in vitro and divided into control group,PQ group and PQ+SKF96365 group.The control group did not take any treatments,the cells of PQ group were given 800μM PQ for 24 h while the cells of PQ+SKF96365 group were given 10μM SKF96365 before given 800μM PQ for 24 h.The activation of SOCE-associated proteins and the translocation of NFATc1 after PQ poisoning was detected by immunofluorescence.A total of forty C57 BL/6 mice were divided randomly into control group,PQ group,SKF96365 group and PQ+SKF96365 group,with 10 mice in each group.The mice of PQ group and PQ+SKF96365 group were given PQ(20 mg/kg)by intraperitoneal injection and the mice of control group were given the equal volume of sterile saline solution instead;The mice of SKF96365 group and PQ+SKF96365 group were given SKF96365(10 mg/kg)once a day for 3 days.The pathological changes of lung tissues and the distribution of collagen fibers were observed by HE and Masson staining.Results:Compared with control group,PQ treated cells exhibit STIM1 aggregation and enhanced membrane distribution of ORAI1 and TRPC1.NFATc1 is significant translocated from cytosol into nucleus after PQ poisoning.The inhibitor of SOCE can decrease the expression and the translocation of NFATc1.Compared with the control group,the alveolar structure of PQ poisoned mice was destroyed and a large area of collagen fiber deposition can be observed.Compared with the PQ group,the alveolar structure of PQ+SKF96365 exhibited is maintained relatively complete intact and the deposition of collagen fiber was largely decreased.Conclusion:PQ poisoning can activate SOCE and promote the occurrence of pulmonary fibrosis by increasing the translocation of NFATc1 from cytosol into nucleus to activate downstream transcription.
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