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1,25-Dihydroxyvitamin D3 preserves intestinal epithelial barrier function from TNF-α induced injury via suppression of NF-kB p65 mediated MLCK-P-MLC signaling pathway.
Biochem Biophys Res Commun. 2015 May 08; 460(3):873-8.BB

Abstract

Substantial studies have demonstrated the protective effect of 1,25-Dihydroxyvitamin D3 (1,25(OH)2D3) on intestinal barrier function, but the mechanisms are not fully illustrated. In this study, the effect of 1,25(OH)2D3 on TNF-α induced barrier dysfunction was further investigated in Caco-2 cell monolayers. The barrier function of Caco-2 monolayers was evaluated by measuring trans-epithelial electrical resistance (TEER) and FITC-Dextran 40,000 Da (FD-40) trans-membrane flux. ZO-1 and Occludin were chosen as markers of the localization of tight junction (TJ) proteins for immunofluorescence. The expression of MLCK and phosphorylation level of myosin light chain (MLC) were measured by immunoblotting. The activation of NF-kB p65 was analyzed by EMSA and immunofluorescence. The results suggest that 1,25(OH)2D3 preserves intestinal epithelial barrier function from TNF-α induced injury via suppression of NF-kB p65 mediated activation of MLCK-P-MLC signaling pathway.

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Publisher Full Text (DOI)

Authors+Show Affiliations

Division of General Surgery, Peking University First Hospital, Peking University, 8 Xi ShiKu Street, Beijing 100034, People's Republic of China.

Division of General Surgery, Peking University First Hospital, Peking University, 8 Xi ShiKu Street, Beijing 100034, People's Republic of China.

Division of General Surgery, Peking University First Hospital, Peking University, 8 Xi ShiKu Street, Beijing 100034, People's Republic of China.

Division of General Surgery, Peking University First Hospital, Peking University, 8 Xi ShiKu Street, Beijing 100034, People's Republic of China.

Division of General Surgery, Peking University First Hospital, Peking University, 8 Xi ShiKu Street, Beijing 100034, People's Republic of China.

Division of General Surgery, Peking University First Hospital, Peking University, 8 Xi ShiKu Street, Beijing 100034, People's Republic of China.

Division of General Surgery, Peking University First Hospital, Peking University, 8 Xi ShiKu Street, Beijing 100034, People's Republic of China.

Division of Gastroenterology, Peking University First Hospital, Peking University, 8 Xi ShiKu Street, Beijing 100034, People's Republic of China.

Division of General Surgery, Peking University First Hospital, Peking University, 8 Xi ShiKu Street, Beijing 100034, People's Republic of China.

Division of General Surgery, Peking University First Hospital, Peking University, 8 Xi ShiKu Street, Beijing 100034, People's Republic of China. Electronic address: wangpengyuan2014@126.com.

MeSH

Base SequenceCaco-2 CellsCalcitriolDNA PrimersHumansIntestinal MucosaMyosin Light ChainsMyosin-Light-Chain KinaseNF-kappa BReal-Time Polymerase Chain ReactionSignal TransductionTumor Necrosis Factor-alpha

Pub Type(s)

Journal Article

Language

eng

PubMed ID

25838204

Citation

Chen, Shanwen, et al. "1,25-Dihydroxyvitamin D3 Preserves Intestinal Epithelial Barrier Function From TNF-α Induced Injury Via Suppression of NF-kB P65 Mediated MLCK-P-MLC Signaling Pathway." Biochemical and Biophysical Research Communications, vol. 460, no. 3, 2015, pp. 873-8.

Chen S, Zhu J, Chen G, et al. 1,25-Dihydroxyvitamin D3 preserves intestinal epithelial barrier function from TNF-α induced injury via suppression of NF-kB p65 mediated MLCK-P-MLC signaling pathway. Biochem Biophys Res Commun. 2015;460(3):873-8.

Chen, S., Zhu, J., Chen, G., Zuo, S., Zhang, J., Chen, Z., Wang, X., Li, J., Liu, Y., & Wang, P. (2015). 1,25-Dihydroxyvitamin D3 preserves intestinal epithelial barrier function from TNF-α induced injury via suppression of NF-kB p65 mediated MLCK-P-MLC signaling pathway. Biochemical and Biophysical Research Communications, 460(3), 873-8. https://doi.org/10.1016/j.bbrc.2015.03.125

Chen S, et al. 1,25-Dihydroxyvitamin D3 Preserves Intestinal Epithelial Barrier Function From TNF-α Induced Injury Via Suppression of NF-kB P65 Mediated MLCK-P-MLC Signaling Pathway. Biochem Biophys Res Commun. 2015 May 8;460(3):873-8. PubMed PMID: 25838204.

* Article titles in AMA citation format should be in sentence-case

TY - JOUR T1 - 1,25-Dihydroxyvitamin D3 preserves intestinal epithelial barrier function from TNF-α induced injury via suppression of NF-kB p65 mediated MLCK-P-MLC signaling pathway. AU - Chen,Shanwen, AU - Zhu,Jing, AU - Chen,Guowei, AU - Zuo,Shuai, AU - Zhang,Junling, AU - Chen,Ziyi, AU - Wang,Xin, AU - Li,Junxia, AU - Liu,Yucun, AU - Wang,Pengyuan, Y1 - 2015/03/31/ PY - 2015/03/07/received PY - 2015/03/23/accepted PY - 2015/4/4/entrez PY - 2015/4/4/pubmed PY - 2015/8/22/medline KW - 1,25(OH)2D3 KW - MLCK KW - TNF-α KW - Tight junction (TJ) KW - Trans-epithelial electrical resistance (TEER) SP - 873 EP - 8 JF - Biochemical and biophysical research communications JO - Biochem Biophys Res Commun VL - 460 IS - 3 N2 - Substantial studies have demonstrated the protective effect of 1,25-Dihydroxyvitamin D3 (1,25(OH)2D3) on intestinal barrier function, but the mechanisms are not fully illustrated. In this study, the effect of 1,25(OH)2D3 on TNF-α induced barrier dysfunction was further investigated in Caco-2 cell monolayers. The barrier function of Caco-2 monolayers was evaluated by measuring trans-epithelial electrical resistance (TEER) and FITC-Dextran 40,000 Da (FD-40) trans-membrane flux. ZO-1 and Occludin were chosen as markers of the localization of tight junction (TJ) proteins for immunofluorescence. The expression of MLCK and phosphorylation level of myosin light chain (MLC) were measured by immunoblotting. The activation of NF-kB p65 was analyzed by EMSA and immunofluorescence. The results suggest that 1,25(OH)2D3 preserves intestinal epithelial barrier function from TNF-α induced injury via suppression of NF-kB p65 mediated activation of MLCK-P-MLC signaling pathway. SN - 1090-2104 UR - https://www.unboundmedicine.com/medline/citation/25838204/125_Dihydroxyvitamin_D3_preserves_intestinal_epithelial_barrier_function_from_TNF_α_induced_injury_via_suppression_of_NF_kB_p65_mediated_MLCK_P_MLC_signaling_pathway_ DB - PRIME DP - Unbound Medicine ER -