OpenLB
Open Library of Bioscience
Advanced Search
Journal of cellular physiology
07, 2020;235 (7-8): 5525-5540.

Astragalus polysaccharides alleviates LPS-induced inflammation via the NF-κB/MAPK signaling pathway.

Na Dong, Xinran Li, Chenyu Xue, Lei Zhang, Chensi Wang, Xinyao Xu, Anshan Shan
Author Information
  1. Na Dong: Laboratory of Molecular Nutrition and Immunity, Northeast Agricultural University, Harbin, P.R. China.
  2. Xinran Li: Laboratory of Molecular Nutrition and Immunity, Northeast Agricultural University, Harbin, P.R. China. ORCID
  3. Chenyu Xue: Laboratory of Molecular Nutrition and Immunity, Northeast Agricultural University, Harbin, P.R. China.
  4. Lei Zhang: Laboratory of Molecular Nutrition and Immunity, Northeast Agricultural University, Harbin, P.R. China.
  5. Chensi Wang: Laboratory of Molecular Nutrition and Immunity, Northeast Agricultural University, Harbin, P.R. China.
  6. Xinyao Xu: Laboratory of Molecular Nutrition and Immunity, Northeast Agricultural University, Harbin, P.R. China.
  7. Anshan Shan: Laboratory of Molecular Nutrition and Immunity, Northeast Agricultural University, Harbin, P.R. China.
PMID: 32037545 DOI: 10.1002/jcp.29452

Abstract

Early weaning usually causes intestinal disorders, enteritis, and diarrhea in young animals and human infants. Astragalus polysaccharides (APS) possesses anti-inflammatory activity. To study the anti-inflammatory mechanisms of APS and its potential effects on intestinal health, we performed an RNA sequencing (RNA-seq) study in lipopolysaccharide (LPS)-stimulated porcine intestinal epithelial cells (IPEC-J2) in vitro. In addition, LPS-stimulated BALB/c mice were used to study the effects of APS on intestinal inflammation in vivo. The results from the RNA-seq analysis show that there were 107, 756, and 5 differentially expressed genes in the control versus LPS, LPS versus LPS+APS, and control versus LPS+APS comparison groups, respectively. The results of Kyoto Encyclopedia of Genes and Genomes enrichment analysis indicated that the mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) signaling pathways play significant roles in the regulation of inflammatory factors and chemokine expression by APS. Further verification of the above two pathways by using western blot and immunofluorescence analysis revealed that the gene expression levels of the phosphorylated p38 MAPK, ERK1/2, and NF-κB p65 were inhibited by APS, while the expression of IκB-α protein was significantly increased (p < .05), indicating that APS inhibits the production of inflammatory factors and chemokines by the inhibition of activation of the MAPK and NF-κB inflammatory pathways induced by LPS stimulation. Animal experiments further demonstrated that prefeeding APS in BALB/c mice can alleviate the expression of the jejunal inflammatory factors interleukin 6 (IL-6), IL-Iβ, and tumor necrosis factor-α induced by LPS stimulation and improve jejunal villus morphology.

Keywords

Astragalus polysaccharides IPEC-J2 Intestinal inflammation MAPK NF-κB RNA-Seq

References

  1. Abraham, C., & Medzhitov, R. (2011). Interactions between the host innate immune system and microbes in inflammatory bowel disease. Gastroenterology, 140(6), 1729-1737.
  2. Afonina, I. S., Tynan, G. A., Logue, S. E., Cullen, S. P., Bots, M., Lüthi, A. U., … Lavelle, E. C. (2011). Granzyme B-dependent proteolysis acts as a switch to enhance the proinflammatory activity of IL-1α. Molecular Cell, 44(2), 265-278.
  3. Andrews, C., Mclean, M. H., & Durum, S. K. (2018). Cytokine tuning of intestinal epithelial function. Frontiers in Immunology, 9, 1270.
  4. Blander, J. M. (2016). Death in the intestinal epithelium-Basic biology and implications for inflammatory bowel disease. FEBS Journal, 283(14), 2720-2730.
  5. Camps, M., Nichols, A., & Arkinstall, S. (2000). Dual specificity phosphatases: A gene family for control of MAP kinase function. FASEB Journal, 14(1), 6-16.
  6. Cario, E., Rosenberg, I. M., Brandwein, S. L., Beck, P. L., Reinecker, H. C., & Podolsky, D. K. (2000). Lipopolysaccharide activates distinct signaling pathways in intestinal epithelial cell lines expressing toll-like receptors. Journal of Immunology, 164, 966-972.
  7. Cho, E. J., Shin, J. S., Noh, Y. S., Cho, Y. W., Hong, S. J., Park, J. H., … Lee, K. T. (2011). Anti-inflammatory effects of methanol extract of Patrinia scabiosaefolia in mice with ulcerative colitis. Journal of Ethnopharmacology, 136(3), 428-435.
  8. Choi, J. H., Chung, K. S., Jin, B. R., Cheon, S. Y., Nugroho, A., Roh, S. S., & An, H. J. (2017). Anti-inflammatory effects of an ethanol extract of Aster glehni via inhibition of NF-κB activation in mice with DSS-induced colitis. Food & Function, 8(7), 2611-2620.
  9. Cox, C. M., Lu, R., Salcin, K., & Wilson, J. M. (2018). The endosomal protein endotubin is required for enterocyte differentiation. Cellular & Molecular Gastroenterology & Hepatology, 5(2), 145-156.
  10. Das, T., Chen, Z., Hendriks, R. W., & Kool, M. (2018). A20/Tumor necrosis factor α-induced protein 3 in immune cells controls development of autoinflammation and autoimmunity: Lessons from mouse models. Frontiers in Immunology, 9, 104.
  11. Deuring, J. J., De, H. C., Kuipers, E. J., Peppelenbosch, M. P., & Cj, V. D. W. (2013). The cell biology of the intestinal epithelium and its relation to inflammatory bowel disease. International Journal of Biochemistry & Cell Biology, 45(4), 798-806.
  12. Diomede, F., Zingariello, M., Cavalcanti, M. F. X. B., Merciaro, I., Pizzicannella, J., Isla, N. D., … Trubiani, O. (2017). MyD88/ERK/NFkB pathways and pro-inflammatory cytokines release in periodontal ligament stem cells stimulated by Porphyromonas gingivalis. European Journal of Histochemistry, 61(2).
  13. Dixon, L. J., Kabi, A., Nickerson, K. P., & Mcdonald, C. (2015). Combinatorial effects of diet and genetics on inflammatory bowel disease pathogenesis. Inflammatory Bowel Diseases, 21(4), 912-922.
  14. Günther, C., Buchen, B., He, G. W., Hornef, M., Torow, N., Neumann, H., … Bleich, A. (2015). Caspase-8 controls the gut response to microbial challenges by Tnf-α-dependent and independent pathways. Gut, 64(4), 601-610.
  15. Griffith, J. W., Sokol, C. L., & Luster, A. D. (2014). Chemokines and chemokine receptors: Positioning cells for host defense and immunity. Annual Review of Immunology, 32(1), 659-702.
  16. Gringhuis, S. I., Kaptein, T. M., Wevers, B. A., Mesman, A. W., & Geijtenbeek, T. B. H. (2014). Fucose-specific DC-SIGN signalling directs T helper cell type-2 responses via IKKε- and CYLD-dependent Bcl3 activation. Nature Communications, 5, 3898.
  17. Ho, Y. C., Lee, S. S., Yang, M. L., Huang-Liu, R., Lee, C. Y., Li, Y. C., & Kuan, Y. H. (2017). Zerumbone reduced the inflammatory response of acute lung injury in endotoxin-treated mice via Akt-NFκB pathway. Chemico-biological interactions, 271, 9-14.
  18. Hughes, C. E., & Rjb, N. (2018). A guide to chemokines and their receptors. FEBS Journal, 285, 2944-2971.
  19. Hung, K. S., Hsiao, C. C., Pai, T. W., Hu, C. H., Tzou, W. S., Wang, W. D., & Chen, Y. R. (2018). Functional enrichment analysis based on long noncoding RNA associations. BMC Systems Biology, 12(4), 45.
  20. Jiang, J. B., Qiu, J. D., Yang, L. H., Jun-Ping, H. E., Smith, G. W., & Hong-Quan, L. I. (2010). Therapeutic effects of astragalus polysaccharides on inflammation and synovial apoptosis in rats with adjuvant-induced arthritis. International Journal of Rheumatic Diseases, 13(4), 396-405.
  21. Jijon, H. B., Buret, A., Hirota, C. L., Hollenberg, M. D., & Beck, P. L. (2012). The EGF receptor and HER2 participate in TNF-α-dependent MAPK activation and IL-8 secretion in intestinal epithelial cells. Mediators of Inflammation, 2012(4), 207398. 2012,(2012-09-5).
  22. Jin, M., Zhao, K., Huang, Q., & Shang, P. (2014). Structural features and biological activities of the polysaccharides from Astragalus membranaceus. International Journal of Biological Macromolecules, 64(2), 257-266.
  23. Keyse, S. M. (1998). Protein phosphatases and the regulation of MAP kinase activity. Seminars in Cell & Developmental Biology, 9(2), 143-152.
  24. Kommadath, A., Bao, H., Arantes, A. S., Plastow, G. S., Tuggle, C. K., Bearson, S. M., … Stothard, P. (2014a). Gene co-expression network analysis identifies porcine genes associated with variation in Salmonella shedding. BMC Genomics, 15(1), 452.
  25. Kommadath, A., Hua, B., Arantes, A. S., Plastow, G. S., Tuggle, C. K., Bearson, S. M., … Stothard, P. (2014b). Gene co-expression network analysis identifies porcine genes associated with variation in Salmonella shedding. BMC Genomics, 15(1), 452. 15,1(2014-06-09).
  26. Lee, A., Qiao, Y., Grigoriev, G., Chen, J., Parkmin, K. H., Park, S. H., … Kalliolias, G. D. (2013). Tumor necrosis factor α induces sustained signaling and a prolonged and unremitting inflammatory response in rheumatoid arthritis synovial fibroblasts. Arthritis & Rheumatism, 65(4), 928-938.
  27. Li, T. T., Ogino, S., & Qian, Z. R. (2014). Toll-like receptor signaling in colorectal cancer: Carcinogenesis to cancer therapy. World Journal of Gastroenterology, 20(47), 17699-17708.
  28. Liu, F., Li, G., Wen, K. T., Cao, D., Zhang, Y., & Yuan, L. (2010). Porcine small intestinal epithelial cell line (IPEC-J2) of rotavirus infection as a new model for the study of innate immune responses to rotaviruses and probiotics. Viral Immunology, 23(2), 135-149.
  29. Liu, Q., Li, A., Yu, S., Qin, S., Han, N., Pestell, R. G., … Wu, K. (2018). DACH1 antagonizes CXCL8 to repress tumorigenesis of lung adenocarcinoma and improve prognosis. Journal of Hematology & Oncology, 11(1), 53.
  30. Luissint, A. C., Parkos, C. A., & Nusrat, A. (2016). Inflammation and the intestinal barrier: Leukocyte-epithelial cell interactions, cell junction remodeling, and mucosal repair. Gastroenterology, 151(4), 616-632.
  31. Lynn, A. M., Harmsen, W. S., Tremaine, W. J., Bazerbachi, F., Dayyeh, B. K. A., & Loftus, E. V. (2018). Su1887-Impact of obesity on future IBD-related complications in a population-based cohort of Crohn's disease (CD) and ulcerative colitis (UC) patients. Gastroenterology, 154(6), S-620-S-621.
  32. Maeda, A., Bandow, K., Kusuyama, J., Kakimoto, K., Ohnishi, T., Miyawaki, S., & Matsuguchi, T. (2015). Induction of CXCL2 and CCL2 by pressure force requires IL-1β-MyD88 axis in osteoblasts. Bone, 74, 76-82.
  33. Mendis, M., Leclerc, E., & Simsek, S. (2016). Arabinoxylan hydrolyzates as immunomodulators in Caco-2 and HT-29 colon cancer cell lines. Food & Function, 8(1).
  34. Miao, E. A., Rajan, J. V., & Aderem, A. (2011). Caspase-1-induced pyroptotic cell death. Immunological Reviews, 243(1), 206-214.
  35. Ng, P. W., Iha, H., Iwanaga, Y., Bittner, M., Chen, Y., Jiang, Y., … Jeang, K. T. (2001). Genome-wide expression changes induced by HTLV-1 Tax: Evidence for MLK-3 mixed lineage kinase involvement in Tax-mediated NF-kappaB activation. Oncogene, 20(33), 4484-4496.
  36. Peterson, L. W., & Artis, D. (2014). Intestinal epithelial cells: Regulators of barrier function and immune homeostasis. Nature Reviews Immunology, 14(3), 141-153.
  37. Salvo, R. E., Alonso, C. C., Pardo, C. C., Casado, B. M., & Vicario, M. (2015). The intestinal barrier function and its involvement in digestive disease. Revista Espanola De Enfermedades Digestivas, 107(11), 686.
  38. Sun, S., Zheng, K., Zhao, H., Lu, C., Liu, B., Yu, C., … He, X. (2014). Regulatory effect of Astragalus polysaccharides on intestinal intraepithelial γδT cells of tumor bearing mice. Molecules, 19(9), 15224-15236.
  39. Tanaka, T., Narazaki, M., & Kishimoto, T. (2014). IL-6 in inflammation, immunity, and disease. Cold Spring Harbor Perspectives in Biology, 6(10), a016295.
  40. Turner, J. R. (2009). Intestinal mucosal barrier function in health and disease. Nature Reviews Immunology, 9(11), 799-809.
  41. Vande, W. L., Van, O. N., Jacques, P., Fossoul, A., Verheugen, E., Vogel, P., … Van, L. G. (2014). Negative regulation of the NLRP3 inflammasome by A20 protects against arthritis. Nature, 512(7512), 69-73.
  42. Vasagiri, N., & Kutala, V. K. (2014). Structure, function, and epigenetic regulation of BNIP3: A pathophysiological relevance. Molecular Biology Reports, 41(11), 7705-7714.
  43. Wang, J., Li, G. R., Tan, B. E., Xiong, X., Kong, X. F., Xiao, D. F., … Kim, S. W. (2015). Oral administration of putrescine and proline during the suckling period improves epithelial restitution after early weaning in piglets. Journal of Animal Science, 93(4), 1679-1688.
  44. Wang, X., Shen, J., Li, S., Zhi, L., Yang, X., & Yao, J. (2014). Sulfated Astragalus polysaccharide regulates the inflammatory reaction in LPS-infected broiler chicks. International Journal of Biological Macromolecules, 69(8), 146-150.
  45. Xiao, Z., Liu, L., Tao, W., Pei, X., Wang, G., & Wang, M. (2018). Clostridium tyrobutyricum protect intestinal barrier function from LPS-induced apoptosis via P38/JNK signaling pathway in IPEC-J2 cells. Cellular Physiology & Biochemistry, 46(5), 1779-1792.
  46. Xiong, X., Yang, H., Tan, B., Yang, C., Wu, M., Liu, G., … Wang, J. (2015). Differential expression of proteins involved in energy production along the crypt-villus axis in early-weaning pig small intestine. Ajp Gastrointestinal & Liver Physiology, 309(4), 229-237. ajpgi.00095.02015.
  47. Xu, C. T., Meng, S. Y., & Pan, B. R. (2004). Drug therapy for ulcerative colitis. World Journal of Gastroenterology, 10(16), 2311-2317.
  48. Xue, H., Gan, F., Qian, G., Hu, J., Hao, S., Xu, J., … Huang, K. (2017). Astragalus polysaccharides attenuate PCV2 infection by inhibiting endoplasmic reticulum stressin vivoandin vitro. Scientific Reports, 7, 40440.
  49. Yang, L. P., Shen, J. G., Xu, W. C., Li, J., & Jiang, J. Q. (2013). Secondary metabolites of the genus Astragalus: Structure and biological-activity update. Chemistry & Biodiversity, 10(6), 1004-1054.
  50. Yang, M., Lin, H. B., Gong, S., Chen, P. Y., Geng, L. L., Zeng, Y. M., & Li, D. Y. (2014a). Effect of Astragalus polysaccharides on expression of TNF-α, IL-1β and NFATc4 in a rat model of experimental colitis. Cytokine, 70(2), 81-86.
  51. Yeh, Y. C., Yang, C. P., Lee, S. S., Horng, C. T., Chen, H. Y., Cho, T. H., … Kuan, Y. H. (2016). Acute lung injury induced by lipopolysaccharide is inhibited by wogonin in mice via reduction of Akt phosphorylation and RhoA activation. Journal of Pharmacy & Pharmacology, 68(2), 257-263.
  52. Yuan, Y., Sun, M., & Li, K. -S. (2009). Astragalus mongholicus polysaccharide inhibits lipopolysaccharide-induced production of TNF-α and interleukin-8. World Journal of Gastroenterology, 15(29), 3676-3680.
  53. Zhou, C., Liu, Z., Jiang, J., Yu, Y., & Zhang, Q. (2012). Differential gene expression profiling of porcine epithelial cells infected with three enterotoxigenic Escherichia coli strains. BMC Genomics, 13(1), 330.
  54. Zhou, M., He, J., Shen, Y., Zhang, C., Wang, J., & Chen, Y. (2017). New frontiers in genetics, gut microbiota, and immunity: A Rosetta stone for the pathogenesis of inflammatory bowel disease. BioMed Research International, 2017(9. 8201672-17.
  55. Zimmerman, N. P., Vongsa, R. A., Wendt, M. K., & Dwinell, M. B. (2010). Chemokines and chemokine receptors in mucosal homeostasis at the intestinal epithelial barrier in inflammatory bowel disease. Inflammatory Bowel Diseases, 14(7), 1000-1011.

MeSH Term

Animals
Astragalus Plant
Gene Expression Regulation
Humans
Inflammation
Interleukin-1beta
Interleukin-6
Lipopolysaccharides
MAP Kinase Signaling System
Mice
NF-KappaB Inhibitor alpha
NF-kappa B
Phosphorylation
RNA-Seq
eIF-2 Kinase
p38 Mitogen-Activated Protein Kinases

Chemicals

Interleukin-1beta
Interleukin-6
Lipopolysaccharides
NF-kappa B
NF-KappaB Inhibitor alpha
eIF-2 Kinase
p38 Mitogen-Activated Protein Kinases
Journal Article Research Support, Non-U.S. Gov't

Word Cloud

Created with Highcharts 10.0.0APSLPSintestinalMAPKNF-κBinflammatoryexpressionAstragaluspolysaccharidesstudyinflammationanalysisversuspathwaysfactorsanti-inflammatoryeffectsRNA-seqIPEC-J2BALB/cmiceresultscontrolLPS+APSproteinsignalinginducedstimulationjejunalEarlyweaningusuallycausesdisordersenteritisdiarrheayounganimalshumaninfantspossessesactivitymechanismspotentialhealthperformedRNAsequencinglipopolysaccharide-stimulatedporcineepithelialcellsvitroadditionLPS-stimulatedusedvivoshow1077565differentiallyexpressedgenescomparisongroupsrespectivelyKyotoEncyclopediaGenesGenomesenrichmentindicatedmitogen-activatedkinasenuclearfactor-κBplaysignificantrolesregulationchemokineverificationtwousingwesternblotimmunofluorescencerevealedgenelevelsphosphorylatedp38ERK1/2p65inhibitedIκB-αsignificantlyincreasedp <05indicatinginhibitsproductionchemokinesinhibitionactivationAnimalexperimentsdemonstratedprefeedingcanalleviateinterleukin6IL-6IL-Iβtumornecrosisfactor-αimprovevillusmorphologyalleviatesLPS-inducedviaNF-κB/MAPKpathwayIntestinalRNA-Seq

Similar Articles

Cited By