Glycerolipid Biosynthesis in Isolated Rat Intestinal Epithelial Cells

Intestinal epithelial cells were prepared from fasted rats by dispersion with collagenase (EC 3.4.24.3). The structural and metabolic integrity of the cells was verified by electron microscopy, a high percentage of Trypan Blue exclusion, a low degree of release of lactate dehydrogenase (EC 1.1.1.27) in the medium, and by the retention of sensitivity to agents known to modify metabolic and transport activity in everted sacs of intestinal mucosa. The isolated intestinal epithelial cells were used to study glycerolipid biosynthesis from glucose, glycerol, 2-monoacylglycerol, and free fatty acids. The cells actively incorporated the labeled precursors into glycerolipids without specific cofactor requirements. Addition of fatty acids stimulated the incorporation of both glucose and glycerol into triacylglycerols and glycerophospholipids, the greatest effect being observed with palmitate. The stimulation of monoacylglycerol acylation appeared to depend on both the nature of the monoacylglycerol and fatty acid supplied. Stereospecific analyses of the diacylglycerols formed from 2-monoacylglycerols and free fatty acids showed that 1,2-diacyl-sn-glycerols (62–70%) were the major and that 2,3-diacyl-sn-glycerols (30–38%) the minor intermediates in triacylglycerol biosynthesis. The data indicate that isolated intestinal epithelial cells exhibit a total capacity of glycerolipid synthesis and a stereochemical course of reaction which is comparable to that observed for triacylglycerol formation in everted sacs of intestinal mucosa, but much less specific than that seen in microsomal preparations of intestinal mucosa.

Download Full-text

Enterococcus faecium PNC01 isolated from the intestinal mucosa of chicken as an alternative for antibiotics to reduce feed conversion rate in broiler chickens

Microbial Cell Factories ◽

10.1186/s12934-021-01609-z ◽

2021 ◽

Vol 20 (1) ◽

Author(s):

Yang He ◽

Xuan Liu ◽

Yuanyang Dong ◽

Jiaqi Lei ◽

Koichi Ito ◽

...

Keyword(s):

Epithelial Cells ◽

Intestinal Mucosa ◽

Conversion Rate ◽

Broiler Chickens ◽

Intestinal Epithelial Cells ◽

Relative Length ◽

Intestinal Epithelial ◽

Feed Conversion ◽

Broiler Production ◽

Feed Conversion Rate

Abstract Background The development and utilization of probiotics had many environmental benefits for replacing antibiotics in animal production. Bacteria in the intestinal mucosa have better adhesion to the host intestinal epithelial cells compared to bacteria in the intestinal contents. In this study, lactic acid bacteria were isolated from the intestinal mucosa of broiler chickens and investigated as the substitution to antibiotic in broiler production. Results In addition to acid resistance, high temperature resistance, antimicrobial sensitivity tests, and intestinal epithelial cell adhesion, Enterococcus faecium PNC01 (E. faecium PNC01) was showed to be non-cytotoxic to epithelial cells. Draft genome sequence of E. faecium PNC01 predicted that it synthesized bacteriocin to perform probiotic functions and bacteriocin activity assay showed it inhibited Salmonella typhimurium from invading intestinal epithelial cells. Diet supplemented with E. faecium PNC01 increased the ileal villus height and crypt depth in broiler chickens, reduced the relative length of the cecum at day 21, and reduced the relative length of jejunum and ileum at day 42. Diet supplemented with E. faecium PNC01 increased the relative abundance of Firmicutes and Lactobacillus, decreased the relative abundance of Bacteroides in the cecal microbiota. Conclusion E. faecium PNC01 replaced antibiotics to reduce the feed conversion rate. Furthermore, E. faecium PNC01 improved intestinal morphology and altered the composition of microbiota in the cecum to reduce feed conversion rate. Thus, it can be used as an alternative for antibiotics in broiler production to avoid the adverse impact of antibiotics by altering the gut microbiota. Graphic Abstract

Download Full-text

LC/ESI/MS Analysis of Saturated and Unsaturated Fatty Acids in Rat Intestinal Epithelial Cells

Current Drug Metabolism ◽

10.2174/138920006779010584 ◽

2006 ◽

Vol 7 (8) ◽

pp. 929-937 ◽

Cited By ~ 18

Author(s):

Seon Hwa Lee ◽

Caterina Pettinella ◽

Ian Blair

Keyword(s):

Fatty Acids ◽

Epithelial Cells ◽

Unsaturated Fatty Acids ◽

Intestinal Epithelial Cells ◽

Intestinal Epithelial ◽

Esi Ms ◽

Ms Analysis

Download Full-text

Diacylglycerol Biosynthesis in Everted Sacs of Rat Intestinal Mucosa

Canadian Journal of Biochemistry ◽

10.1139/o75-162 ◽

1975 ◽

Vol 53 (11) ◽

pp. 1170-1183 ◽

Cited By ~ 17

Author(s):

W. C. Breckenridge ◽

A. Kuksis

Keyword(s):

Fatty Acids ◽

Phosphatidic Acid ◽

Thin Layer ◽

Free Fatty Acids ◽

Thin Layer Chromatography ◽

Intestinal Mucosa ◽

Gas Liquid Chromatography ◽

Acid Pathway ◽

Layer Chromatography ◽

Everted Sacs

The molecular specificity in the biosynthesis of diacylglycerols by rat intestinal mucosa was examined by means of radioactive markers, thin-layer chromatography with silver nitrate and gas-liquid chromatography with radioactivity monitoring. Bile salt micelles of alternately labeled monoacylglycerols and free fatty acids were incubated with everted sacs of intestinal mucosa for various periods of time and the diacylglycerols were isolated by solvent extraction and thin-layer chromatography. Stereospecific analyses of the X-1,2-diacylglycerols labeled from 2-monoacylgiycerols showed that the sn-1,2-isomers (45–55%) were slightly in excess of the sn-2,3-isomers (34–45%) with the X-1,3-diacylglycerols accounting for the rest of the radioactivity (5–10%). This suggests that racemic diacylglycerols may be intermediates in the resynthesis of dietary fat in rat intestinal mucosa. Detailed analyses of the molecular species of the sn-1,2-diacylglycerols labeled from free fatty acids revealed that 10–45% of the total did not contain the acid present in the 2-monoacylglycerol supplied, and therefore had originated from the phosphatidic acid pathway. These findings are at variance with those obtained in isolated microsomes, which have suggested an inhibition of the phosphatidic acid pathway by monoacylglycerols as well as have given evidence of an exclusive synthesis of sn-1,2-diacylglycerols from 2-monoacylglycerols.

Download Full-text