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[维生素D受体对缺氧环境下结肠细胞黏膜屏障蛋白的影响]

[Effects of Vitamin D Receptor on Mucosal Barrier Proteins in Colon Cells under Hypoxic Environment].

作者信息

Wang Zheng, Yang Hong, Jin Meng, Zhang Hui Min, Chen Xuan Fu, Wu Mei Xu, Guo Ming Yue, Huang Chang Zhi, Qian Jia Ming

机构信息

Department of Gastroenterology,PUMC Hospital,CAMS and PUMC,Beijing 100730,China.

State Key Laboratory of Molecular Oncology,Cancer Hospital,CAMS and PUMC,Beijing 100021,China.

出版信息

Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2019 Aug 30;41(4):506-511. doi: 10.3881/j.issn.1000-503X.11243.

Abstract

To investigate the expressions of mucosal barrier proteins in colon cell line DLD-1 under hypoxic environment and its mechanism. Methods After DLD-1 cells were treated separately with hypoxia(l% O),vitamin D(100 nmol/L),or vitamin D plus hypoxia for 48 hours,the expressions of vitamin D receptor(VDR),tight junction proteins zonula occludens-1(ZO-1),occludin,Claudin-1,and adherent junction protein(E-cadherin)were determined by Western blot.Stable VDR knock-down(Sh-VDR)DLD-1 cell line and control DLD-1 cell line were established by lentivirus package technology and the protein expressions after hypoxia treatment were detected. Results Compared with control group,the expressions of occludin,Claudin-1,and VDR increased significantly after hypoxia treatment(all <0.001).In addition to the protein expressions of occludin,Claudin-1 and VDR,the expressions of ZO-1 and E-cadherin were also obviously higher in vitamin D plus hypoxia group than in single vitamin D treatment group(all <0.001).After hypoxia treatment,Sh-VDR cell line showed significantly decreased expressions of ZO-1(<0.001),occludin(<0.05),Claudin-1(<0.01)and E-cadherin(<0.001)when compared with untreated Sh-VDR cell line. Conclusion VDR acts as a regulator for the expressions of intestinal mucosal barrier proteins under hypoxia environment in DLD-1 colon cell line,indicating that VDR pathway may be another important protective mechanism for gut barrier in low-oxygen environment.

摘要

探讨低氧环境下结肠癌细胞系DLD-1中黏膜屏障蛋白的表达及其机制。方法 将DLD-1细胞分别用低氧(1% O₂)、维生素D(100 nmol/L)或维生素D联合低氧处理48小时后,采用蛋白质免疫印迹法检测维生素D受体(VDR)、紧密连接蛋白闭合蛋白1(ZO-1)、闭锁小带蛋白(occludin)、Claudin-1及黏附连接蛋白(E-钙黏蛋白)的表达。通过慢病毒包装技术构建稳定敲低VDR(Sh-VDR)的DLD-1细胞系和对照DLD-1细胞系,并检测低氧处理后的蛋白表达。结果 与对照组相比,低氧处理后occludin、Claudin-1及VDR的表达显著增加(均P<0.001)。维生素D联合低氧组除occludin、Claudin-1及VDR蛋白表达外,ZO-1和E-钙黏蛋白的表达也明显高于单纯维生素D处理组(均P<0.001)。低氧处理后,Sh-VDR细胞系与未处理的Sh-VDR细胞系相比,ZO-1(P<0.001)、occludin(P<0.05)、Claudin-1(P<0.01)及E-钙黏蛋白(P<0.001)的表达显著降低。结论 在DLD-1结肠癌细胞系低氧环境下,VDR作为肠黏膜屏障蛋白表达的调节因子,提示VDR途径可能是低氧环境中肠道屏障的另一种重要保护机制。

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