Jejunal Alkaline Phosphatase Activity Research Articles

Clostridium butyricum relieves diarrhea by enhancing digestive function, maintaining intestinal barrier integrity, and relieving intestinal inflammation in weaned piglets

The current study aimed to determine how Clostridium butyricum (C. butyricum) relieved diarrhea by evaluating its effects on intestinal digestive enzyme activity, barrier integrity, inflammatory responses, and oxidative stress in weaned piglets. A total of 360 weaned piglets that were 25 days old (180 males and 180 females) were randomly assigned to 5 groups in 6 pens, with 12 pigs per pen, and offered the same antibiotic-free corn-soybean-type basal diet for 30 d. The groups were as follows: no additives (control), basal diet + 250 mg/kg C. butyricum (CB1), basal diet + 500 mg/kg C. butyricum (CB2), basal diet + 1,000 mg/kg C. butyricum (CB3), and basal diet + 2,000 mg/kg C. butyricum (CB4). The incidence of diarrhea decreased quadratically as the amount of C. butyricum supplementation was increased, and this decrease was minimized in the CB2 group. Compared with those in the control group, the duodenal amylase and lipase activities were increased quadratically and maximized in the CB2 group, and the jejunal lipase activity was increased cubically in the CB1 group, while the pancreatic trypsin activity was increased linearly and maximized in the CB4 group. The serum concentration of D-lactic acid responded to the increases in the levels of C. butyricum in a quadratic manner; the minimum level was observed in the CB2 group. Jejunal zonula occludens-1 expression displayed linear and quadratic trends following C. butyricum supplementation, with the maximum response observed in the CB3 group. Quadratic and cubic increases were observed in jejunal alkaline phosphatase activity in the CB3 group. The jejunal IL-10 concentration increased, while the jejunal vascular endothelial growth factor concentration decreased quadratically in response to dietary C. butyricum supplementation in the CB2 group. No significant differences among the groups were observed in terms of the jejunal mucosal concentrations of oxidation-associated enzymes and products. In conclusion, dietary supplementation with appropriate doses of C. butyricum could relieve diarrhea in weaned piglets by enhancing digestive enzyme activity, maintaining intestinal barrier integrity, and relieving intestinal inflammation.

Piglets Are Vulnerable to Intestinal Inflammation and Infection Because of Expression of Low‐Affinity Jejunal Alkaline Phosphatase Activities

Intestinal alkaline phosphatase (IAP) plays a pivotal role in protecting gut health through catalyzing the hydrolytic dephosphorylation of the lipid A moiety of endotoxin lipopolysaccharides (LPS) and other emblematic members of pathogen‐associated‐molecular patterns (PAMPs) such as ATP, ultimately detoxifying endotoxins and preventing gut dysbiosis and disorders. Characterization of IAP hydrolytic activities, including enzyme affinity (Km) and the maximum enzyme activity (Vmax), has been reported in pigs and other mammalian species by using the chromogenic synthetic substrate P‐nitrophenyl phosphate (pNPP) rather than using relevant physiological substrates such as ATP and LPS under the physiological pH conditions. This study was conducted to determine kinetics of young porcine jejunal alkaline phosphatase in hydrolyzing the dephosphorylation of ATP, the lipid A moiety of LPS and pNPP at pH 7.4 and 37 °C. Jejunal samples were collected from four 10‐day old Yorkshire pigs and were pulverized to be homogenous under liquid N2. The LPS kinetic experiments were conducted with 11 gradient concentrations of LPS, isolated from E. coli 0111:B4, ranging 0 – 5 mg/mL in incubation media. The ATP kinetic experiments were conducted with 11 gradient concentrations of ATP, ranging 0 – 5 mM in incubation media. Free phosphorus (Pi) release was measured by using the PiColorLock detection reagent (Innova Biosciences). Contributions of the non‐specific background Pi associated with ATP and LPS in the time course and the kinetic experiments were corrected. The pNPP kinetic experiments were conducted with 16 gradient concentrations of pNPP, ranging 0 – 6 mM in incubation media. All enzymatic incubations were conducted with 19 – 27 μg of the homogenized porcine jejunal protein. The kinetics (parameter estimates ± SE, P < 0.05) of young porcine jejunal alkaline phosphatase activities were obtained, including Vmax values of 2.5 ± 0.4 ug/mg protein·min (n = 11), 1.1 ± 0.2 ug/mg protein·min (n = 11), and 0.12 ± 0. 01 umol/mg protein·min (n = 16); and Km values of 1.255 ± 0.498 mM (n = 11), 2.388 ± 0.794 mg/mL (n = 11), and 0.145 ± 0.036 mM (n = 16), for the hydrolysis of ATP, LPS and pNPP, respectively. Our results reveal the young porcine IAP Km value 41 times higher in comparison with reported Km value of human IAP in dephosphorylating ATP, 3 times higher when compared with reported human IAP dephosphorylating LPS, and 5 times higher when compared with reported human IAP hydrolyzing pNPP. Results of this study show that young porcine jejunal IAP activity affinity is several fold lower in detoxifying endotoxin LPS the other emblematic PAMPs member ATP in comparison with humans, suggesting that young pigs are susceptible to gut inflammation and infection.Support or Funding InformationNSERC Discovery Program of Canada and Metagen Enzyme Corporation

Long-term effect of altered nutrition induced by litter size manipulation and cross-fostering in suckling male rats on development of obesity risk and health complications

We investigated the long-term effect of pre-weaning nutrition on positive and/or adverse regulation of obesity risk and health complications in male Sprague-Dawley rats. Two experimental models were used in the present work: (1) To induce postnatal over- or normal nutrition, the litter size was adjusted to 4 (small litters-SL) and to 10 pups (normal litters-NL) in the nest, (2) in suckling pups at day 10, we used cross-fostering to identify the effect of altered dietary environment on their future body fat regulation, food intake, blood pressure, and the duodenal and jejunal alkaline phosphatase activity. After weaning, these control (NL, SL) and cross-fostered (NL-SL, SL-NL) groups were exposed to standard laboratory diet. On day 50, the SL in comparison with NL rats became heavier and displayed enhanced adiposity accompanied by significantly increased systolic blood pressure (19%) and duodenal (16%) and jejunal (21%) alkaline phosphatase (AP) activity. The impact of pre-weaning over-nutrition of NL-SL pups was associated with long-lasting positive effect on obesity. In contrast, SL-NL rats submitted until weaning to the opposite normalized feeding condition on day 50 showed significantly decreased fat deposition (21%), systolic blood pressure (20%), and AP activity in duodenum and jejunum (14%). These results contribute to a better understanding of how early-acquired dietary habits determine the attenuation or prevention of obesity development in later life and can provide some benefit for optimizing the future dietary strategies in young and adult obese individuals.

Effects of diets supplemented with organic acids and nucleotides on growth, immune responses and digestive tract development in weaned pigs

Sixty-eight (Experiment 1, 46 days feeding) and sixteen (Experiment 2, 21 days feeding) 21-days-old weaned pigs were allotted to four dietary treatments including control, 0.6% organic acids (OA), 0.1% nucleotides (NA) and 0.6% OA plus 0.1% NA for determining the dietary effects. In Experiment 1, OA enhanced peripheral blood mononuclear cells proliferation on day 28 and 46. The plasma immunoglobulin (Ig) A level was elevated by OA (p < 0.06) and NA (p < 0.07), respectively. In Experiment 2, NA increased plasma IgM level, and had an interactive effect with OA on ileal Peyer's patches and mesenteric lymph node lymphocyte proliferation, bile and plasma IgA levels, and jejunal crypt depth. NA elevated gastric pepsin and jejunal alkaline phosphatase activities, however, decreased ileal aminopeptidase N, sucrase or maltase activity. These results suggest that OA and NA have synergistically enhanced the gut-associated lymphocyte responses and NA modulates the digestive tract development of weaned pigs.

Effects of Medium-Chain Triglycerides on Brush Border Membrane-Bound Enzyme Activity in Rat Small Intestine

The effects of various types of dietary fat on brush border membrane-bound enzymes in rat intestinal mucosa were examined. Four groups of five rats were pair-fed defined diets for 10 d. The control group was fed a diet containing 57% sucrose and 2% corn oil as a fixed carbohydrate reference; the three experimental groups received diets containing 57% sucrose and 2% corn oil plus 13% fat in the form of medium-chain triglycerides (MCT) or long-chain triglycerides (LCT) (either lard as a highly saturated fat or corn oil as a highly unsaturated fat). Feeding LCT compared to the control diet, decreased sucrase activity in mucosal brush borders of the duodenum and jejunum. In these segments of MCT-fed rats, sucrase activity was similar to that in the control animals. In another experiment, measuring immunoreactive sucrase-isomaltase in jejunal brush border membranes revealed that feeding a high corn oil diet, but not a high MCT diet, led to a reduction in the sucrase catalytic activity per unit weight of enzyme protein, suggesting that the degradation status of sucrase-isomaltase might be altered by the different types of dietary fats. With MCT feeding, jejunal alkaline phosphatase activity was enhanced to a large extent compared to the activity in other groups. Feeding MCT, compared to lard or corn oil, also increased microvillus phospholipids of the jejunal mucosa. These results suggest that MCT, unlike LCT, do not suppress the activity of mucosal microvillus membrane enzymes in rat small intestine.

Effect of chronic ethanol and food deprivation on intestinal villus morphology and brush border membrane content of lipid and marker enzymes.

Brush border membranes (BBM) were isolated from the jejunum and ileum of control, ad libitum (CAL); control, food-restricted (CFR); control, weight gain (CWG); and ethanol-fed (EF) rabbits. Jejunal alkaline phosphatase activity was similar among control groups, but higher in CAL than EF animals. Sucrase activity was higher in EF and CWG animals than in CAL and CFR. The alkaline phosphatase/sucrase ratio was lower in EF than control animals. Ileal enzyme marker activity was similar among EF and control animals. Sucrase (S) activity was lower in the ileum than in the jejunum. Jejunal free fatty acid and phospholipid/cholesterol (PL/C) were lower in EF than control animals, whereas ileal lipid content was generally similar among all animal groups. Total phospholipid content was similar between sites, but the cholesterol and free fatty acid content were lower in the ileum than the jejunum. The phospholipid/cholesterol ratio was increased only in the ileum of EF animals. The amount of lecithin was decreased in the jejunal BBM of EF animals resulting in a decreased choline/amine phospholipid ratio as compared with control animals. The ileal phospholipid composition was similar among all groups. A large increase in villus height is observed in the jejunum of EF animals. Villus surface area and mucosal surface area are altered with ethanol feeding and food deprivation. Thus, (i) there is a gradient of S and cholesterol between the BBM of jejunum and ileum; (ii) changes in food intake are associated with changes in the morphology as well as the enzyme marker and lipid content of BBM.(ABSTRACT TRUNCATED AT 250 WORDS)