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Zinc recovers altered intestinial ion‐transport and barrier function caused by Shigella infection in T84 cells (902.9)
Author(s) -
Kazi Mirajul,
Paramita Sarkar,
Sheikh Irshad,
Chakraborty Subhra
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.902.9
Subject(s) - paracellular transport , tight junction , ussing chamber , claudin , shigella flexneri , transcellular , barrier function , chemistry , shigella , secretion , biophysics , microbiology and biotechnology , caco 2 , biology , biochemistry , in vitro , permeability (electromagnetism) , escherichia coli , membrane , gene
Zinc (Zn) has been shown to improve diarrhea. However, mechanism linking Zn to improve inflammatory diarrhea caused by Shigella sp. is not known. We tested that Zn restore paracellular and transcellular ion transport by modulating tight junction protein expression and chloride secretion, altered by Shigella infection. Human colonic epithelial cells, T84 were grown onto transwell inserts for measurements of permeability to dextrans, transepithelial electrical resistance (TER), short‐circuit current (Isc), and dilution potential (DP) in Ussing chamber. Cells infected with Shigella flexneri 2a caused reduction of TER by 40%, DP [4 ± 1.2 vs. 1.5 ± 0.3 mV] which was prevented in presence of Zn. An increase in paracellular flux of 4kDa dextran [2.1± 0.6 vs. 0.8 ± 0.2 µg/3.5hr/cm 2 ] with respect to 70kDa was evident while in the presence of Zn, flux of 4kDa dextran was reversed by 52%. Immunofluorescence study revealed removal of claudin 1 and 2 from tight junction complex correlated with the loss of TER and paracellular flux. Studies with Ussing chambers demonstrated reduced cAMP stimulated electrogenic ion transport in infected cells. Zn application prevented this effect. Our results suggest that in T84 cells, Shigella infection caused (1) altered barrier function (2) altered Cl secretion. Zn application reversed these transport functions. These conclude that Zn may have beneficial role in inflammatory diarrhea. Grant Funding Source : Supported by Dept. of Biotechnology, Govt. of India