Abstract
Conflicting results have been reported that butyrate in normal piglets leads either to an increase or to a decrease of jejunal villus length, implying a possible effect on the proliferation of enterocytes. No definitive study was found for the biological effects of butyrate in porcine jejunal epithelial cells. The present study used IPEC-J2 cells, a non-transformed jejunal epithelial line to evaluate the direct effects of sodium butyrate on cell proliferation, cell cycle regulation, and apoptosis. Low concentrations (0.5 and 1 mM) of butyrate had no effect on cell proliferation. However, at 5 and 10 mM, sodium butyrate significantly decreased cell viability, accompanied by reduced levels of p-mTOR and PCNA protein. Sodium butyrate, in a dose-dependent manner, induced cell cycle arrest in G0/G1 phase and reduced the numbers of cells in S phase. In addition, relative expression of p21, p27, and pro-apoptosis bak genes, and protein levels of p21Waf1/Cip1, p27Kip1, cyclinD3, CDK4, and Cleave-caspase3 were increased by higher concentrations of sodium butyrate (1, 5, 10 mM), and the levels of cyclinD1 and CDK6 were reduced by 5 and 10 mM butyrate. Butyrate increased the phosphorylated form of the signaling molecule p38 and phosphorylated JNK. In conclusion, the present in vitro study indicated that sodium butyrate inhibited the proliferation of IPEC-J2 cells by inducing cell cycle arrest in the G0/G1 phase of cell cycles and by increasing apoptosis at high concentrations.
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References
Basu A, Haldar S (1998) The relationship between BcI2, Bax and p53: consequences for cell cycle progression and cell death. Mol Hum Reprod 4:1099–1109
Berschneider, HM (1989) Development of normal cultured small intestinal epithelial cell lines which transport Na and Cl. Gastroenterology: A41
Biagi G, Piva A, Moschini M, Vezzali E, Roth FX (2007) Performance, intestinal microflora, and wall morphology of weanling pigs fed sodium butyrate. J Anim Sci 85:1184–1191
Chen TH, Chen WM, Hsu KH, Kuo CD, Hung SC (2007) Sodium butyrate activates ERK to regulate differentiation of mesenchymal stem cells. Biochem Biophys Res Commun 355:913–918
Chopin V, Toillon RA, Jouy N, Le Bourhis X (2004) P21WAF1/CIP1 is dispensable for G1 arrest, but indispensable for apoptosis induced by sodium butyrate in MCF-7 breast cancer cells. Oncogene 23:21–29
Davie JR (2003) Inhibition of histone deacetylase activity by butyrate. J Nutr 133:2485–2493
Fernandes-Alnemri T, Litwack G, Alnemri ES (1994) CPP32, a novel human apoptotic protein with homology to Caenorhabditis elegans cell death protein Ced-3 and mammalian interleukin-1 beta-converting enzyme. J Biol Chem 269:30761–30764
Hanczakowska E, Niwińska B, Grela ER, Węglarzy K, Okoń K (2014) Effect of dietary glutamine, glucose and/or sodium butyrate on piglet growth, intestinal environment, subsequent fattener performance, and meat quality. Czech Journal of Animal Science 59:460–470
Hofmanová J, Vaculová A, Koubková Z, Hýžd’alová M, Kozubík A (2009) Human fetal colon cells and colon cancer cells respond differently to butyrate and PUFAs. Mol Nutr Food Res 53:102–113
Khan S, Jena GB (2014) Protective role of sodium butyrate, a HDAC inhibitor on beta-cell proliferation, function and glucose homeostasis through modulation of p38/ERK MAPK and apoptotic pathways: study in juvenile diabetic rat. Chem Biol Interact 213:1–12
Kien CL, Blauwiekel R, Bunn JY, Jetton TL, Frankel WL, Holst JJ (2007) Cecal infusion of butyrate increases intestinal cell proliferation in piglets. J Nutr 137:916–922
Kotunia A, Wolin’ ski J, Laubitz D, Jurkowska M, Rome V, Guilloteau P, Zabielski R (2004) Effect of sodium butyrate on the small intestine development in neonatal piglets fed by artificial sow. J Physiol Pharmacol 55:59–68
Kranenburn O, Scharnhorst V, Van der Eb AJ, Zantema A (1995) Inhibition of cyclin-dependent kinase activity triggers neuronal differentiation of mouse neuroblastoma cells. J Cell Biol 131:227–234
Lacorn M, Goerke M, Claus R (2010) Inulin-coated butyrate increases ileal MCT1 expression and affects mucosal morphology in the porcine ileum by reduced apoptosis. J Anim Physiol Anim Nutr (Berl) 94:670–676
Lallemand F, Courilleau D, Sabbah M, Redeuilh G, Mester J (1996) Direct inhibition of the expression of cyclin D1 gene by sodium butyrate. Biochem Biophys Res Commun 229:163–169
Le Gall M, Gallois M, Seve B, Louveau I, Holst JJ, Oswald IP, Guilloteau P (2009) Comparative effect of orally administered sodium butyrate before or after weaning on growth and several indices of gastrointestinal biology of piglets. Br J Nutr 102:1285–1296
Li CJ, Elsasser TH (2005) Butyrate-induced apoptosis and cell cycle arrest in bovine kidney epithelial cells: involvement of caspase and proteasome pathways. J Anim Sci 83:89–97
Li Y, Liu J, Tang LJ, Shi YW, Ren W, Hu WX (2007) Apoptosis induced by lycorine in KM3 cells is associated with the G0/G1 cell cycle arrest. Oncol Rep 17:377–384
Mentschel J, Claus R (2003) Increased butyrate formation in the pig colon by feeding raw potato starch leads to a reduction of colonocyte apoptosis and a shift to the stem cell compartment. Metabolism 52:1400–1405
Mohana Kumar B, Song HJ, Cho SK, Balasubramanian S, Choe SY, Rho GJ (2007) Effect of histone acetylation modification with sodium butyrate, a histone deacetylase inhibitor, on cell cycle, apoptosis, ploidy and gene expression in porcine fetal fibroblasts. J Reprod Dev 53:903–913
Nofrarias M, Martinez-Puig D, Pujols J, Majó N, Pérez JF (2007) Longterm intake of resistant starch improves colonic mucosal integrity and reduces gut apoptosis and blood immune cells. Nutrition 23:861–870
Pierce KM, Callan JJ, McCarthy P, O’Doherty JV (2007) The interaction between lactose level and crude protein concentration on piglet post-weaning performance, nitrogen metabolism, selcted faecal microbial populations and faecal volatile fatty acid concentrations. Anim Feed Sci Technol 132:267–282
Roediger WE (1980) The colonic epithelium in ulcerative colitis: an energy-deficiency disease? Lancet 316:712–715
Sakata T (1989) Stimulatory effect of short-chain fatty acids on epithelial cell proliferation of isolated and denervated jejunal segment of the rat. Scand J Gastroenterol 24:886–890
Scheppach W, Dusel G, Kuhn T, Loges C, Karch H, Bartram HP, Richter F, Christl SU, Kasper H (1996) Effect of L-glutamine and n-butyrate on the restitution of rat colonic mucosa after acid induced injury. Gut 38:878–885
Siavoshian S, Segain JP, Kornprobst M, Bonnet C, Cherbut C, Galmiche JP, Blottiere HM (2000) Butyrate and trichostatin A effects on the proliferation/differentiation of human intestinal epithelial cells: induction of cyclin D3 and p21 expression. Gut 46:507–514
Wang JF, Chen X, Wang ZX, Dong SH, Lai ZW (2005) Effect of sodium butyrate on the structure of the small intestine mucous epithelium of weaning piglets. Chin J Vet Sci Technol 35:298–301
Yaku K, Enami Y, Kurajyo C, Matsui-Yuasa I, Konishi Y, Kojima-Yuasa A (2012) The enhancement of phase 2 enzyme activities by sodium butyrate in normal intestinal epithelial cells is associated with Nrf2 and p53. Mol Cell Biochem 370:7–14
Zuo L, Lu M, Zhou Q, Wei W, Wang Y (2013) Butyrate suppresses proliferation and migration of RKO colon cancer cells though regulating endocan expression by MAPK signaling pathway. Food Chem Toxicol 62:892–900
Acknowledgements
We gratefully acknowledge W. B. Currie (Cornell University, Ithaca, NY) for suggestions on presentation. This work was supported by grants from the National Natural Science Foundation of China (Grant No. 31402086); Guangdong international science and technology cooperation program (2014A050503049), National Basic Research Program (973) of China (No. 2013CB127301, 2013CB127304), and the Science and Technology Program of Guangdong Province (2016LM1080, 2016A020210041, 2013A061401020).
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Editor: Tetsuji Okamoto
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Qiu, Y., Ma, X., Yang, X. et al. Effect of sodium butyrate on cell proliferation and cell cycle in porcine intestinal epithelial (IPEC-J2) cells. In Vitro Cell.Dev.Biol.-Animal 53, 304–311 (2017). https://doi.org/10.1007/s11626-016-0119-9
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DOI: https://doi.org/10.1007/s11626-016-0119-9