The beneficial effects of intestinal microorganisms rely on short-chain fatty acids (SCFAs), which serve as a crucial material basis.However, disruptions in the intestinal microecology may lead to an absence or excessive absorption of SCFAs, and had adverse effects on the body.Therefore, intestinal epithelial Caco-2 cells were used to establish a model for absorbing mixed SCFAs in the intestine to explore the regulatory effect and possible mechanism of Lactobacillus plantarum on the absorption of SCFAs.Results showed that the surface of Caco-2 cell monolayer was covered with well-developed microvilli, with excellent compactness and integrity at the 21 days of cultivation.The survival rate of intestinal epithelial Caco-2 cells was higher when the mixed SCFAs (acetic acid∶propionic acid∶butyric acid=3∶1∶1, molar ratio) in the model were 5.0 mmol/L.The absorption of acetic acid, propionic acid, and butyric acid by cells in the model was significantly promoted by L.plantarum f28, f2, and f5 (P<0.05), and the promotion of L.plantarum f5 on cells absorption of acetic acid was significantly higher than that of other strains (P<0.05).While L.plantarum f16 showed a significant inhibitory effect (P<0.05) on acetic acid absorption, but its promotion of the absorption of propionic acid and butyric acid was significantly higher than that of the other strains (P<0.05).Meanwhile, L.plantarum f19 significantly inhibited cellular absorption of butyric acid (P<0.05).Both L.plantarum f2 and f19 significantly upregulated the protein and gene expression levels of monocarboxylate transporter 1 (MCT1) in the cells (P<0.05), while L.plantarum f28 and f5 played a significant downregulation role (P<0.05).Additionally, L.plantarum f28, f5, f16, and f19 significantly upregulated the protein and gene expression levels of sodium-coupled monocarboxylate transporter-1 (SMCT1) in the cells (P<0.05).Additionally, L.plantarum f16 significantly upregulated the protein and gene expression levels of Na+/H+ exchanger3 (NHE3) (P<0.05), while L.plantarum f2, f5, and fl9 had a significant down-regulating effect (P<0.05).In conclusion, the L.plantarum could regulate their absorption of SCFAs by up-regulating or down-regulating the expression level of SCFAs transporters in the intestinal epithelial Caco-2 cells, providing theoretical basis for improving the bioavailability of SCFAs in the body and related product development and application.
[1] ALEXANDER C, SWANSON K S, FAHEY G C, et al. Perspective: Physiologic importance of short-chain fatty acids from nondigestible carbohydrate fermentation[J]. Advances in Nutrition, 2019, 10(4):576-589.
[2] WŁODARCZYK J, PŁOSKA M, PŁOSKI K, et al. The role of short-chain fatty acids in inflammatory bowel diseases and colorectal cancer[J]. Postepy Biochemii, 2021, 67(3):223-230.
[3] LI X, SHIMIZU Y, KIMURA I. Gut microbial metabolite short-chain fatty acids and obesity[J]. Bioscience of Microbiota, Food and Health, 2017, 36(4):135-140.
[4] FERNÁNDEZ J, REDONDO-BLANCO S, GUTIÉRREZ-DEL-RÍO I, et al. Colon microbiota fermentation of dietary prebiotics towards short-chain fatty acids and their roles as anti-inflammatory and antitumour agents: A review[J]. Journal of Functional Foods, 2016, 25:511-522.
[5] MA J Y, PIAO X S, MAHFUZ S, et al. The interaction among gut microbes, the intestinal barrier and short chain fatty acids[J]. Animal Nutrition (Zhongguo Xu Mu Shou Yi Xue Hui), 2021, 9:159-174.
[6] DALILE B, VAN OUDENHOVE L, VERVLIET B, et al. The role of short-chain fatty acids in microbiota-gut-brain communication[J]. Nature Reviews. Gastroenterology & Hepatology, 2019, 16(8):461-478.
[7] HUR K Y, LEE M S. Gut microbiota and metabolic disorders[J]. Diabetes & Metabolism Journal, 2015, 39(3):198-203.
[8] BLOEMEN J G, VENEMA K, VAN DE POLL M C, et al. Short chain fatty acids exchange across the gut and liver in humans measured at surgery[J]. Clinical Nutrition, 2009, 28(6):657-661.
[9] MODESTO A, CAMERON N R, VARGHESE C, et al. Meta-analysis of the composition of human intestinal gases[J]. Digestive Diseases and Sciences, 2022, 67(8):3842-3859.
[10] YANG T, RICHARDS E M, PEPINE C J, et al. The gut microbiota and the brain-gut-kidney axis in hypertension and chronic kidney disease[J]. Nature Reviews Nephrology, 2018, 14:442-456.
[11] SUN Y Y, JING D D. Relationship between Helicobacter pylori infection and gastrointestinal microecology[J]. World Chinese Journal of Digestology, 2020, 28(24):1261-1265.
[12] GOPAL E, FEI Y J, SUGAWARA M, et al. Expression of slc5a8 in kidney and its role in Na(+)-coupled transport of lactate[J]. The Journal of Biological Chemistry, 2004, 279(43):44522-44532.
[13] BORTHAKUR A, GILL R K, HODGES K, et al. Enteropathogenic Escherichia coli inhibits butyrate uptake in Caco-2 cells by altering the apical membrane MCT1 level[J]. American Journal of Physiology. Gastrointestinal and Liver Physiology, 2006, 290(1): G30-G35.
[14] GILL R K, BORTHAKUR A, HODGES K, et al. Mechanism underlying inhibition of intestinal apical Cl/OH exchange following infection with enteropathogenic E. coli[J]. The Journal of Clinical Investigation, 2007, 117(2):428-437.
[15] PENG L Y, HE Z J, CHEN W, et al. Effects of butyrate on intestinal barrier function in a Caco-2 cell monolayer model of intestinal barrier[J]. Pediatric Research, 2007, 61(1):37-41.
[16] OVERBY H B, FERGUSON J F. Gut Microbiota-Derived Short-Chain Fatty Acids Facilitate Microbiota: Host Cross talk and Modulate Obesity and Hypertension[J]. Current Hypertension Reports, 2021, 23(2):8.
[17] ZHANG J, ZHAO X, JIANG Y Y, et al. Antioxidant status and gut microbiota change in an aging mouse model as influenced by exopolysaccharide produced by Lactobacillus plantarum YW11 isolated from Tibetan kefir[J]. Journal of Dairy Science, 2017, 100(8):6025-6041.
[18] BORTHAKUR A, ANBAZHAGAN A N, KUMAR A, et al. The probiotic Lactobacillus plantarum counteracts TNF-{alpha}-induced downregulation of SMCT1 expression and function[J]. American Journal of Physiology. Gastrointestinal and Liver Physiology, 2010, 299(4): G928-G934.
[19] CHEN D W, CHEN C M, QU H X, et al. Screening of Lactobacillus strains that enhance SCFA uptake in intestinal epithelial cells[J]. European Food Research and Technology, 2021, 247(5):1049-1060.
[20] KUMAR A, ALREFAI W A, BORTHAKUR A, et al. Lactobacillus acidophilus counteracts enteropathogenic E. coli-induced inhibition of butyrate uptake in intestinal epithelial cells[J]. American Journal of Physiology. Gastrointestinal and Liver Physiology, 2015, 309(7): G602-G607.
[21] NASIRI G, AZIMIRAD M, GOUDARZI H, et al. The inhibitory effects of live and UV-killed Akkermansia muciniphila and its derivatives on cytotoxicity and inflammatory response induced by Clostridioides difficile RT001 in vitro[J]. International Microbiology, 2024, 27(2):393-409.
[22] 范晨雨. 促肠上皮细胞吸收短链脂肪酸乳酸菌的筛选及其作用机制[D]. 扬州: 扬州大学, 2020.
FAN C Y. Screening and mechanism of lactic acid bacteria promoting intestinal epithelial cells to absorb short-chain fatty acids[D].Yangzhou: Yangzhou University, 2020.
[23] STERLING D, ALVAREZ B V, CASEY J R. The extracellular component of a transport metabolon. Extracellular loop 4 of the human AE1 Cl-/HCO-3 exchanger binds carbonic anhydrase IV[J]. The Journal of Biological Chemistry, 2002, 277(28):25239-25246.
[24] NOACH A B J, KUROSAKI Y, BLOM-ROOSEMALEN M C M, et al. Cell-polarity dependent effect of chelation on the paracellular permeability of confluent caco-2 cell monolayers[J]. International Journal of Pharmaceutics, 1993, 90(3):229-237.
[25] MING X, THAKKER D R. Role of basolateral efflux transporter MRP4 in the intestinal absorption of the antiviral drug adefovir dipivoxil[J]. Biochemical Pharmacology, 2010, 79(3):455-462.
[26] 周慧敏. 利用Caco-2细胞模型研究肠道可吸收乳清肽的结构特征[D]. 南京: 南京师范大学, 2015.
ZHOU H M. Study on structural characteristics of intestinal absorbable whey peptide by Caco-2 cell model[D]. Nanjing: Nanjing Normal University, 2015.
[27] SANGSAWAD P, ROYTRAKUL S, CHOOWONGKOMON K, et al. Transepithelial transport across Caco-2 cell monolayers of angiotensin converting enzyme (ACE) inhibitory peptides derived from simulated in vitro gastrointestinal digestion of cooked chicken muscles[J]. Food Chemistry, 2018, 251:77-85.
[28] 关溯, 赵立子, 陈杰, 等. Caco-2细胞模型的建立及意义[J]. 山东医药, 2005, 45(26):1-3.
GUAN S, ZHAO L Z, CHEN J, et al. Establishment and value of Caco-2 cell model[J]. Shandong Medical Journal, 2005, 45(26):1-3.
[29] YEE S. In vitro permeability across Caco-2 cells (colonic) can predict in vivo (small intestinal) absorption in man: Fact or myth[J]. Pharmaceutical Research, 1997, 14(6):763-766.
[30] LI M H, LONG X, WAN H J, et al. Monocarboxylate transporter 1 promotes proliferation and invasion of renal cancer cells by mediating acetate transport[J]. Cell Biology International, 2021, 45(6):1278-1287.
[31] 林华林, 秦颖超, 周加义, 等. 丁酸梭菌及其代谢产物对畜禽肠道健康影响的研究进展[J]. 饲料工业, 2019, 40(6):7-12.
LIN H L, QIN Y C, ZHOU J Y, et al. Research progress on the effects of Clostridium butyricum and its metabolites on intestinal health of livestock[J]. Feed Industry, 2019, 40(6):7-12.
[32] BORTHAKUR A, GILL R K, TYAGI S, et al. The probiotic Lactobacillus acidophilus stimulates chloride/hydroxyl exchange activity in human intestinal epithelial cells[J]. The Journal of Nutrition, 2008, 138(7):1355-1359.
[33] HU J, MA L B, NIE Y F, et al. A microbiota-derived bacteriocin targets the host to confer diarrhea resistance in early-weaned piglets[J]. Cell Host & Microbe, 2018, 24(6):817-832.
[34] MUSCARIELLO L, DE SIENA B, MARASCO R. Lactobacillus cell surface proteins involved in interaction with mucus and extracellular matrix components[J]. Current Microbiology, 2020, 77(12):3831-3841.
[35] HOLST B, GLENTING J, HOLMSTRØM K, et al. Molecular switch controlling expression of the mannose-specific adhesin, msa, in Lactobacillus plantarum[J]. Applied and Environmental Microbiology, 2019, 85(10): e02954-e02918.
