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Total polysaccharides of the Sijunzi decoction attenuate tumor necrosis factor-α-induced damage to the barrier function of a Caco-2 cell monolayer via the nuclear factor-κB-myosin light chain kinase-myosin light chain pathway.
World J Gastroenterol. 2018 Jul 14; 24(26):2867-2877.WJ

Abstract

AIM

To explore the protective effects and underlying mechanisms of total polysaccharides of the Sijunzi decoction (TPSJ) on the epithelial barriers in vitro.

METHODS

Caco-2 cell monolayers were treated with or without TPSJ in the presence or absence of TNF-α, and paracellular permeability and transepithelial electrical resistance (TEER) were measured to evaluate the epithelial barrier function. Immunofluorescence and western blotting were respectively used to evaluate the distribution and expression of the tight junction proteins claudin 1, claudin 2, zo3, and occludin in Caco-2 cells. Western blotting was also used to evaluate the cellular expression of myosin light chain (MLC), phosphorylated MLC (pMLC), MLC kinase (MLCK), and nuclear factor (NF)-κB p65.

RESULTS

TPSJ promoted the proliferation of Caco-2 cells and inhibited TNF-α-induced secretion of pro-inflammatory cytokines. Furthermore, TPSJ significantly ameliorated both the reduction of TEER and the increased paracellular permeability observed in tumor necrosis factor (TNF)-α-damaged Caco-2 monolayers. Furthermore, TPSJ remarkably attenuated TNF-α-induced morphological changes, downregulated the expression of claudin 1, claudin 2, zo3, and occludin, and markedly suppressed TNF-α-mediated upregulation of p-MLC and MLCK expression. Finally, TPSJ inhibited the activation and expression of NF-κB p65.

CONCLUSION

Our results demonstrate that TPSJ alleviates the TNF-α-induced impairment of the intestinal epithelial cell barrier function by suppressing NF-κB p65-mediated phosphorylation of MLCK and MLC.

Authors+Show Affiliations

The Second Clinical College (Guangdong Provincial Hospital of Chinese Medicine), Guangzhou University of Chinese Medicine, Guangzhou 510006, Guangdong Province, China.The Second Clinical College (Guangdong Provincial Hospital of Chinese Medicine), Guangzhou University of Chinese Medicine, Guangzhou 510006, Guangdong Province, China.The Second Clinical College (Guangdong Provincial Hospital of Chinese Medicine), Guangzhou University of Chinese Medicine, Guangzhou 510006, Guangdong Province, China.The Second Clinical College (Guangdong Provincial Hospital of Chinese Medicine), Guangzhou University of Chinese Medicine, Guangzhou 510006, Guangdong Province, China.The Second Clinical College (Guangdong Provincial Hospital of Chinese Medicine), Guangzhou University of Chinese Medicine, Guangzhou 510006, Guangdong Province, China.The Second Clinical College (Guangdong Provincial Hospital of Chinese Medicine), Guangzhou University of Chinese Medicine, Guangzhou 510006, Guangdong Province, China. linghan99@gzucm.edu.cn.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

30018481

Citation

Lu, Yue, et al. "Total Polysaccharides of the Sijunzi Decoction Attenuate Tumor Necrosis Factor-α-induced Damage to the Barrier Function of a Caco-2 Cell Monolayer Via the Nuclear factor-κB-myosin Light Chain Kinase-myosin Light Chain Pathway." World Journal of Gastroenterology, vol. 24, no. 26, 2018, pp. 2867-2877.
Lu Y, Li L, Zhang JW, et al. Total polysaccharides of the Sijunzi decoction attenuate tumor necrosis factor-α-induced damage to the barrier function of a Caco-2 cell monolayer via the nuclear factor-κB-myosin light chain kinase-myosin light chain pathway. World J Gastroenterol. 2018;24(26):2867-2877.
Lu, Y., Li, L., Zhang, J. W., Zhong, X. Q., Wei, J. A., & Han, L. (2018). Total polysaccharides of the Sijunzi decoction attenuate tumor necrosis factor-α-induced damage to the barrier function of a Caco-2 cell monolayer via the nuclear factor-κB-myosin light chain kinase-myosin light chain pathway. World Journal of Gastroenterology, 24(26), 2867-2877. https://doi.org/10.3748/wjg.v24.i26.2867
Lu Y, et al. Total Polysaccharides of the Sijunzi Decoction Attenuate Tumor Necrosis Factor-α-induced Damage to the Barrier Function of a Caco-2 Cell Monolayer Via the Nuclear factor-κB-myosin Light Chain Kinase-myosin Light Chain Pathway. World J Gastroenterol. 2018 Jul 14;24(26):2867-2877. PubMed PMID: 30018481.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Total polysaccharides of the Sijunzi decoction attenuate tumor necrosis factor-α-induced damage to the barrier function of a Caco-2 cell monolayer via the nuclear factor-κB-myosin light chain kinase-myosin light chain pathway. AU - Lu,Yue, AU - Li,Leng, AU - Zhang,Jin-Wei, AU - Zhong,Xiao-Qin, AU - Wei,Jian-An, AU - Han,Ling, PY - 2018/03/29/received PY - 2018/05/25/revised PY - 2018/06/09/accepted PY - 2018/7/19/entrez PY - 2018/7/19/pubmed PY - 2018/11/6/medline KW - Inflammatory bowel disease KW - Nuclear factor-κB pathway KW - Tight junction KW - Total polysaccharides of the Sijunzi decoction SP - 2867 EP - 2877 JF - World journal of gastroenterology JO - World J Gastroenterol VL - 24 IS - 26 N2 - AIM: To explore the protective effects and underlying mechanisms of total polysaccharides of the Sijunzi decoction (TPSJ) on the epithelial barriers in vitro. METHODS: Caco-2 cell monolayers were treated with or without TPSJ in the presence or absence of TNF-α, and paracellular permeability and transepithelial electrical resistance (TEER) were measured to evaluate the epithelial barrier function. Immunofluorescence and western blotting were respectively used to evaluate the distribution and expression of the tight junction proteins claudin 1, claudin 2, zo3, and occludin in Caco-2 cells. Western blotting was also used to evaluate the cellular expression of myosin light chain (MLC), phosphorylated MLC (pMLC), MLC kinase (MLCK), and nuclear factor (NF)-κB p65. RESULTS: TPSJ promoted the proliferation of Caco-2 cells and inhibited TNF-α-induced secretion of pro-inflammatory cytokines. Furthermore, TPSJ significantly ameliorated both the reduction of TEER and the increased paracellular permeability observed in tumor necrosis factor (TNF)-α-damaged Caco-2 monolayers. Furthermore, TPSJ remarkably attenuated TNF-α-induced morphological changes, downregulated the expression of claudin 1, claudin 2, zo3, and occludin, and markedly suppressed TNF-α-mediated upregulation of p-MLC and MLCK expression. Finally, TPSJ inhibited the activation and expression of NF-κB p65. CONCLUSION: Our results demonstrate that TPSJ alleviates the TNF-α-induced impairment of the intestinal epithelial cell barrier function by suppressing NF-κB p65-mediated phosphorylation of MLCK and MLC. SN - 2219-2840 UR - https://www.unboundmedicine.com/medline/citation/30018481/Total_polysaccharides_of_the_Sijunzi_decoction_attenuate_tumor_necrosis_factor_α_induced_damage_to_the_barrier_function_of_a_Caco_2_cell_monolayer_via_the_nuclear_factor_κB_myosin_light_chain_kinase_myosin_light_chain_pathway_ L2 - https://www.wjgnet.com/1007-9327/full/v24/i26/2867.htm DB - PRIME DP - Unbound Medicine ER -