scholarly journals Octreotide Alleviates Autophagy by Up-Regulation of MicroRNA-101 in Intestinal Epithelial Cell Line Caco-2

2018 ◽  
Vol 49 (4) ◽  
pp. 1352-1363 ◽  
Author(s):  
Yuling Li ◽  
Su Wang ◽  
Xingjuan Gao ◽  
Ying Zhao ◽  
Yongwei Li ◽  
...  
Keyword(s):  
Tight Junction ◽  
Cell Viability ◽  
Mtor Pathway ◽  
Common Side Effect ◽  
Epithelial Cell Line ◽  
Therapeutic Effects ◽  
Intestinal Epithelial ◽  
Differentiated Cells ◽  
Monolayer Permeability ◽  
Underlying Mechanisms

Background: Intestinal mucositis is a common side-effect after anti-cancer therapy, which may greatly restrict the therapeutic effects. We aimed to explore the functional role of octreotide (OCT) in lipopolysaccharide (LPS)-induced autophagy of human intestinal epithelial cells as well as the underlying mechanisms. Methods: Cell viability and expression of proteins related to autophagy, AMPK and the mTOR pathway in LPS-treated Caco-2 cells were determined by CCK-8 assay and Western blot analysis, respectively. Effects of OCT on LPS-induced alterations as well as miR-101 expression were measured. Then, miR-101 was aberrantly expressed, and whether OCT alleviated LPS-induced autophagy through miR-101 was tested. Next, whether TGF-β-activated kinase 1 (TAK1) was involved in the regulation of miR-101 in LPS-induced autophagy was studied. Effects of OCT on monolayer permeability and tight junction level were analyzed via measuring transepithelial electrical resistance (TEER) and expression of tight junction proteins. Results: LPS reduced cell viability and increased autophagy through activating AMPK and inhibiting the mTOR pathway in Caco-2 cells. OCT alleviated LPS-induced alterations and repressed degradation of autophagosome. Then, we found that OCT affected autophagy through up-regulating miR-101 in LPS-treated cells. Moreover, miR-101-induced inactivation of AMPK and activation of the mTOR pathway in LPS-treated cells were reversed by inhibition of TAK1 phosphorylation. Finally, we found miR-101 was up-regulated in differentiated cells, and OCT protected the monolayer permeability and tight junction level. Conclusion: OCT repressed autophagy through miR-101-mediated inactivation of TAK1, along with inactivation of AMPK and activation of the mTOR pathway in LPS-treated Caco-2 cells.

scholarly journals PSVI-37 Effects of deoxynivalenol (DON) and selected feed additives on viability and tight junction integrity of IPEC-J2 cells

2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 202-202
Author(s):  
Hang Lu ◽  
Kolapo Ajuwon
Keyword(s):  
Tight Junction ◽  
Feed Additives ◽  
Control Treatment ◽  
Epithelial Cell Line ◽  
Intestinal Epithelial ◽  
Negative Effects ◽  
Semipermeable Membranes ◽  
Livestock Feed ◽  
Epithelial Tight Junction ◽  
Porcine Intestinal Epithelial Cell

Abstract Intestinal epithelial cells represent the frontline of host defense against invasion by enteric pathogens and uptake of potentially harmful substances. Mycotoxins are deleterious feed contaminants routinely found in cereal grains used in livestock feed. Development of strategies to eliminate mycotoxins and mitigate their harmful effects on the animal is of major importance to the livestock industry. The objective of the current study was to determine the effects of deoxynivalenol (DON), a common mycotoxin contaminant, and selected prebiotic, probiotic, and essential oil blended feed additives, on viability and integrity in porcine intestinal epithelial cell line (IPEC-J2). IPEC-J2 cells were grown on either plastic or semipermeable membranes until 100% confluence and treated with DON (12.5 uM) with or without additives (0.01 mg/mL Biolex® and 0.0001 mg/mL Encinnate®, Biomatrix Inc.) Use of DON at 12.5 uM decreased the trans-epithelial electrical resistance (TEER) by 37.5% compared to control, and led to 88.7% increase in FITC-dextran permeability after 3 days of culture, but with no negative effects on cell viability. The decrease in TEER by DON was minimized in the presence of Biolex (22.2%) and Encinnate (20.5%) relative to control. Treatment with DON also decreased the mRNA (24.0%) and protein abundance (64.4%) of claudin 3 and this was partially reversed by the additives, 11.5 and 11.0% decrease for claudin 3 mRNA and 58.7 and 56.7% decrease for the protein relative to control for Biolex and Encinnate, respectively. In summary, DON led to impaired epithelial tight junction integrity and this was partially rescued by the additives tested.

Protective Effect of Schisandrin B Against Cyclosporine A-Induced Nephrotoxicity In Vitro and In Vivo

2012 ◽  
Vol 40 (03) ◽  
pp. 551-566 ◽  
Author(s):  
Shaohua Zhu ◽  
Yan Wang ◽  
Meiwan Chen ◽  
Jing Jin ◽  
Yuwen Qiu ◽  
...  
Keyword(s):  
Cell Viability ◽  
Cyclosporine A ◽  
Tubular Epithelial Cell ◽  
Epithelial Cell Line ◽  
Ros Generation ◽  
Therapeutic Effects ◽  
Schisandrin B ◽  
Protective Effects

Schisandrin B (Sch B) is an active ingredient of the fruit of Schisandra chinensis. It has many therapeutic effects arising from its tonic, sedative, antitussive and antiaging activities and is also used in the treatment of viral and chemical hepatitis. The aim of this study was to investigate the protective effects of Sch B on cyclosporine A (CsA)-induced nephrotoxicity in mice and HK-2 cells (a human proximal tubular epithelial cell line). After gavage with Sch B (20 mg/kg) or olive oil (vehicle), mice received CsA (30 mg/kg) by subcutaneous injection once daily for four weeks. Renal function, histopathology, and tissue glutathione (GSH) and malondialdehyde (MDA) levels were evaluated after the last treatment. The effects of Sch B on CsA–induced oxidative damage in HK-2 cells were investigated by measuring cell viability, the release of lactate dehydrogenase (LDH), the level of reactive oxygen species (ROS), and the cellular GSH and ATP concentrations. Cellular apoptosis was assessed by flow cytometry. Treatment with Sch B in CsA-treated mice significantly suppressed the elevation of blood urea nitrogen (BUN) and serum creatinine levels and attenuated the histopathological changes. Additionally, Sch B also decreased renal MDA levels and increased GSH levels in CsA-treated mice. Using an in vitro model, Sch B (2.5, 5 and 10 μM) significantly increased the cell viability and reduced LDH release and apoptosis induced by CsA (10 μM) in HK-2 cells. Furthermore, Sch B increased the intracellular GSH and ATP levels and attenuated CsA-induced ROS generation. In conclusion, Sch B appears to protect against CsA-induced nephrotoxicity by decreasing oxidative stress and cell death.

scholarly journals 56 Effects of mycotoxins and selected feed additives on viability and tight junction integrity of IPEC-J2 cells

2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 37-37
Author(s):  
Hang Lu ◽  
Nathan Horn ◽  
Kola Ajuwon
Keyword(s):  
Gene Expression ◽  
Tight Junction ◽  
Immune Modulation ◽  
Fumonisin B1 ◽  
Feed Additives ◽  
Epithelial Cell Line ◽  
Protein Abundance ◽  
Intestinal Epithelial ◽  
Negative Effects ◽  
Tlr2 Gene

Abstract Mycotoxins are deleterious feed contaminants routinely found in cereal gains used in livestock feed. Development of strategies to eliminate mycotoxins and mitigate their harmful effects on the animal is of major importance to the livestock industry. The objective of the current study was to determine the effects of deoxynivalenol (DON), aflatoxin B1 (AFB1) and fumonisin B1(FB1), and selected prebiotic, probiotic, and essential oil blended feed additives, on viability and integrity in porcine intestinal epithelial cell line (IPEC-J2). IPEC-J2 cells were treated with each individual mycotoxin (DON, AFB1 and FB1) with or without additives (Microsecure (MS), Biolex (BL) and Encinnate (EN), Biomatrix Inc., U.S.A). Mycotoxins significantly decreased the trans-epithelial electrical resistance (TEER) of IPEC-J2 cells compared to control with no negative effects on cell viability at 72h. Biolex and EN increased TEER under a DON challenge. The decrease in TEER by AFB1 was minimized in the presence of MS (9.8%), BL (17.1%) and EN (22.8%) relative to AFB1 (P < 0.0001). The protein abundance of claudin 3 was decreased with mycotoxins (P < 0.0001) and this was partially reversed by the additives relative to control cells with additives. In addition, BL and EN significantly increased claudin 4 protein abundance (P = 0.02) when challenged with AFB1. Encinnate significantly increased TLR2 gene expression when challenged with DON (P < 0.05) with MS and BL had a numerical TLR2 gene expression. A numerical increasing of TLR2 gene expression was also observed with feed additives when cells were challenged with AFB1 and FB1. In summary, mycotoxins led to impaired epithelial tight junction integrity and this was in-part rescued by the additives tested through restoration of cellular integrity and TLR-2 dependent immune modulation.

The Effects of Lauric Acid on IPEC-J2 Cell Differentiation, Proliferation, and Death

2020 ◽  
Vol 20 (7) ◽  
pp. 572-581
Author(s):  
Yuan Yang ◽  
Jin Huang ◽  
Jianzhong Li ◽  
Huansheng Yang ◽  
Yulong Yin
Keyword(s):  
Cell Death ◽  
Cell Differentiation ◽  
Cell Viability ◽  
P38 Mapk ◽  
Lauric Acid ◽  
Cell Apoptosis ◽  
Epithelial Cell Line ◽  
Intestinal Epithelial ◽  
Metabolism Regulation ◽  
Cell Proliferation Inhibition

Background: Lauric acid (LA) has antimicrobial effects and the potential to replace antibiotics in feeds to prevent postweaning diarrhea and increase overall swine productivity. The effects of lauric acid on the intestinal epithelial cells remain unclear. Materials and Methods: This study investigates the effects of LA on pig intestinal epithelial cell line (IPEC-J2) differentiation, proliferation, and death and explores its underlying mechanisms. It was found that 0.25-0.1 mM LA promoted IPEC-J2 cell differentiation. At 1 mM or higher concentrations, it induced IPEC-J2 cell viability decreases, lipid accumulation, cell proliferation inhibition, and cell apoptosis. The cell death induced did not depend on caspase pathways. : The data demonstrated that LA induced the IPEC-J2 cell autophagy and impaired autophagy flux and autophagy plays a role in protecting against LA induced-cell death. p38 MAPK inhibitor SB202190 attenuated LA-reduced IPEC-J2 cell viability. This associated with an increase in autophagy level and a decrease in lipid accumulations and FABPI levels. Conclusion: Conclusion: In summary, LA promoted the IPEC-J2 cell apoptosis depends on the p38 MAPK pathways and may involve autophagy and TG metabolism regulation.

The Effects of Butyric Acid on the Differentiation, Proliferation, Apoptosis, and Autophagy of IPEC-J2 Cells

2020 ◽  
Vol 20 (4) ◽  
pp. 307-317
Author(s):  
Yuan Yang ◽  
Jin Huang ◽  
Jianzhong Li ◽  
Huansheng Yang ◽  
Yulong Yin
Keyword(s):  
Cell Viability ◽  
P38 Mapk ◽  
Butyric Acid ◽  
Cell Apoptosis ◽  
Mapk Pathway ◽  
Mrna Level ◽  
Dependent Manner ◽  
M Phase ◽  
High Concentrations ◽  
Underlying Mechanisms

Background: Butyric acid (BT), a short-chain fatty acid, is the preferred colonocyte energy source. The effects of BT on the differentiation, proliferation, and apoptosis of small intestinal epithelial cells of piglets and its underlying mechanisms have not been fully elucidated. Methods: In this study, it was found that 0.2-0.4 mM BT promoted the differentiation of procine jejunal epithelial (IPEC-J2) cells. BT at 0.5 mM or higher concentrations significantly impaired cell viability in a dose- and time-dependent manner. In addition, BT at high concentrations inhibited the IPEC-J2 cell proliferation and induced cell cycle arrest in the G2/M phase. Results: Our results demonstrated that BT triggered IPEC-J2 cell apoptosis via the caspase8-caspase3 pathway accompanied by excess reactive oxygen species (ROS) and TNF-α production. BT at high concentrations inhibited cell autophagy associated with increased lysosome formation. It was found that BT-reduced IPEC-J2 cell viability could be attenuated by p38 MAPK inhibitor SB202190. Moreover, SB202190 attenuated BT-increased p38 MAPK target DDIT3 mRNA level and V-ATPase mRNA level that were responsible for normal acidic lysosomes. Conclusion: In conclusion, 1) at 0.2-0.4 mM, BT promotes the differentiation of IPEC-J2 cells; 2) BT at 0.5 mM or higher concentrations induces cell apoptosis via the p38 MAPK pathway; 3) BT inhibits cells autophagy and promotes lysosome formation at high concentrations.

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