An Active Fraction of Trillium tschonoskii Promotes the Regeneration of Intestinal Epithelial Cells After Irradiation

Despite significant scientific advances toward the development of safe and effective radiation countermeasures, no drug has been approved for use in the clinic for prevention or treatment of radiation-induced acute gastrointestinal syndrome (AGS). Thus, there is an urgent need to develop potential drugs to accelerate the repair of injured intestinal tissue. In this study, we investigated that whether some fractions of Traditional Chinese Medicine (TCM) have the ability to regulate intestinal crypt cell proliferation and promotes crypt regeneration after radiation. By screening the different supplements from a TCM library, we found that an active fraction of the rhizomes of Trillium tschonoskii Maxim (TT), TT-2, strongly increased the colony-forming ability of irradiated rat intestinal epithelial cell line 6 (IEC-6) cells. TT-2 significantly promoted the proliferation and inhibited the apoptosis of irradiated IEC-6 cells. Furthermore, in a small intestinal organoid radiation model, TT-2 promoted irradiated intestinal organoid growth and increased Lgr5+ intestinal stem cell (ICS) numbers. More importantly, the oral administration of TT-2 remarkably enhanced intestinal crypt cell proliferation and promoted the repair of the intestinal epithelium of mice after abdominal irradiation (ABI). Mechanistically, TT-2 remarkably activated the expression of ICS-associated and proliferation-promoting genes and inhibited apoptosis-related gene expression. Our data indicate that active fraction of TT can be developed into a potential oral drug for improving the regeneration and repair of intestinal epithelia that have intestinal radiation damage.

PSXVI-7 Supplemental effects of Lactobacillus extract postbiotics in prevention of post-weaning diarrhea caused by enterotoxigenic F18+ Escherichia coli in nursery pigs

This study determined the supplemental effects of Lactobacillus extract (LBE) postbiotics on intestinal health and prevention of postweaning diarrhea caused by F18+ Escherichia coli (ETEC) in nursery pigs. Sixty-four weaned pigs (6.6 ± 0.7 kg BW) were allotted in a RCBD to 4 dietary treatments (NC: no-challenge; PC: challenge/no-treat; BMD: challenge/bacitracin; LBE: challenge/LBE 0.2%) and fed diets for 28 d. At d 7, challenged groups were orally inoculated with ETEC (2.4 x 1010 CFU) and NC group received sterile solution. Growth performance was analyzed weekly and pigs were euthanized on d 28 to measure intestinal health. Data were analyzed using the SAS 9.4. During post-challenge period, PC tended to decrease (P = 0.067) ADG (373 to 284 g/d), whereas BMD increased (P < 0.05) ADG (284 to 408 g/d); LBE tended to increase (P = 0.081) ADG (284 to 370 g/d). PC increased fecal score (P < 0.05, 3.4 to 3.9) on d 14, whereas BMD decreased it (P < 0.05, 3.9 to 3.5) on d 21. PC increased (P < 0.05) protein carbonyl (0.76 to 1.12 nmol/mg protein), crypt cell proliferation (28 to 36%), and Helicobacter rodentium (0.4 to 3.7%). However, BMD decreased (P < 0.05) crypt cell proliferation (36 to 32%) and Helicobacter spp. (15.0 to 1.4%); and increased (P < 0.05) villus height (309 to 377 µm), Bifidobacterium boum (0.04 to 2.0%), Pelomonas spp. (1.5 to 8.5%), and Microbacterium ginsengisoli (0.5 to 3.0%). LBE reduced (P < 0.05) crypt cell proliferation (36 to 27%) and Helicobacter rodentium (3.7 to 0.04%); and increased (P < 0.05) Lactobacillus salivarius (0.3 to 4.1%) and Propionibacterium acnes (0.4 to 7.4%). Collectively, ETEC reduced growth performance by adversely affecting microbiota and intestinal health. BMD and LBE improved growth performance by enhancing intestinal health and increasing beneficial microbiota in ETEC challenged pigs.

The murine intestinal stem cells are highly sensitive to the modulation of the T3/TRα1-dependent pathway

The thyroid hormone T3 and its nuclear receptor TRα1 control gut development and homeostasis through the modulation of intestinal crypt cell proliferation. Despite increasing data, in depth analysis on their specific action on intestinal stem cells is lacking.By using ex vivo 3D organoid cultures and molecular approaches we observed early responses to T3 involving the T3-metabolizing enzyme Dio1 and the transporter Mct10, accompanied by a complex response of stem cell- and progenitor-enriched genes. Interestingly, specific TRα1 loss-of-function (inducible or constitutive) was responsible for low ex vivo organoid development and impaired stem cell activity. T3-treatment of animals in vivo not only confirmed the positive action of this hormone on crypt cell proliferation but also demonstrated its key action in modulating i) the number of the stem cells, ii) the expression of their specific markers and iii) the commitment of progenitors into lineage-specific differentiation.In conclusion, T3 treatment or TRα1 modulation has a rapid and strong effect on intestinal stem cells, broadening our perspectives in the study of T3/TRα1-dependent signaling in these cells.

Understanding intestinal health in nursery pigs and the relevant nutritional strategies

In the modern pig production, pigs are weaned at early age with immature intestine. Dietary and environmental factors challenge the intestine, specifically the jejunum, causing inflammation and oxidative stress followed by destruction of epithelial barrier and villus structures in the jejunum. Crypt cell proliferation increases to repair damages in the jejunum. Challenges to maintain the intestinal health have been shown to be related to changes in the profile of mucosa-associated microbiota in the jejunum of nursery pigs. All these processes can be quantified as biomarkers to determine status of intestinal health related to growth potential of nursery pigs. Nursery pigs with impaired intestinal health show reduced ability of nutrient digestion and thus reduced growth. A tremendous amount of research effort has been made to determine nutritional strategies to maintain or improve intestinal health and microbiota in nursery pigs. A large number of feed additives have been evaluated for their effectiveness on improving intestinal health and balancing intestinal microbiota in nursery pigs. Selected prebiotics, probiotics, postbiotics, and other bioactive compounds can be used in feeds to handle issues with intestinal health. Selection of these feed additives should aim modulating biomarkers indicating intestinal health. This review aims to define intestinal health and introduce examples of nutritional approaches to handle intestinal health in nursery pigs.

Dose–response and functional role of whey permeate as a source of lactose and milk oligosaccharides on intestinal health and growth of nursery pigs

Two experiments were conducted to evaluate dose–response and supplemental effects of whey permeate on growth performance and intestinal health of nursery pigs. In experiment (exp.) 1, 1,080 pigs weaned at 6.24 kg body weight (BW) were allotted to five treatments (eight pens/treatment) with increasing levels of whey permeate in three phases (from 10% to 30%, 3% to 23%, and 0% to 9% for phase 1, 2, and 3, respectively) fed until 11 kg BW and then fed a common phase 4 diet (0% whey permeate) until 25 kg BW in a 48-d feeding trial. Feed intake and BW were measured at the end of each phase. In exp. 2, 1,200 nursery pigs at 7.50 kg BW were allotted to six treatments (10 pens/treatment) with increasing levels of whey permeate from 0% to 18.75% fed until 11 kg BW. Feed intake and BW were measured during 11 d. Six pigs per treatment (1 per pens) were euthanized to collect the jejunum to evaluate tumor necrosis factor-alpha, interleukin-8 (IL-8), transforming growth factor-beta 1, mucin 2, histomorphology, digestive enzyme activity, crypt cell proliferation rate, and jejunal mucosa-associated microbiota. Data were analyzed using contrasts in the MIXED procedure and a broken-line analysis using the NLIN procedure of SAS. In exp. 1, increasing whey permeate had a quadratic effect (P < 0.05) on feed efficiency (G:F; maximum: 1.35 at 18.3%) in phase 1. Increasing whey permeate linearly increased (P < 0.05) average daily gain (ADG; 292 to 327 g/d) and G:F (0.96 to 1.04) of pigs in phase 2. In exp. 2, increasing whey permeate linearly increased (P < 0.05) ADG (349 to 414 g/d) and G:F (0.78 to 0.85) and linearly increased (P < 0.05) crypt cell proliferation rate (27.8% to 37.0%). The breakpoint from a broken-line analysis was obtained at 13.6% whey permeate for maximal G:F. Increasing whey permeate tended to change IL-8 (quadratic, P = 0.052; maximum: 223 pg/mg at 10.9%), to decrease Firmicutes:Bacteroidetes (P = 0.073, 1.59 to 1.13), to increase (P = 0.089) Bifidobacteriaceae (0.73% to 1.11%), and to decrease Enterobacteriaceae (P = 0.091, 1.04% to 0.52%) and Streptococcaceae (P = 0.094, 1.50% to 0.71%) in the jejunal mucosa. In conclusion, dietary inclusion of whey permeate increased the growth of nursery pigs from 7 to 11 kg BW. Pigs grew most efficiently with 13.6% whey permeate. Improvement in growth performance is partly attributed to stimulating intestinal immune response and enterocyte proliferation with positive changes in jejunal mucosa-associated microbiota in nursery pigs.

PSIV-22 Supplemental effects of dietary lysophospholipids in lactation diets on sow performance, milk composition, and intestinal health of piglets

Dietary lysophospholipids could enhance nutrient utilization through a structural change of enterocyte membrane with increasing permeability. The objective of this study was to determine supplemental effects of dietary lysophospholipids in lactation diets on sow performance, milk characteristics, and intestinal health of piglets. The 52 pregnant sows were allotted to 2 treatments in randomized complete block design with parity and BW as blocks at d 110 of pregnancy. The treatments were CON (no added lysophospholipids) and LPL (at 0.05% lysophospholipids; Lipidol-Ultra, Pathway Intermediates, Shrewsbury, UK). The lactation diets were formulated to meet or exceed nutrient requirements suggested by NRC (2012). Milk samples from 12 sows per treatment were collected to measure gross energy, protein, fat, fatty acid profile, and immunoglobulins (IgG and IgA) on d 1 and d 18 of lactation. Twelve piglets per treatment were euthanized on d 18 to collect tissues to measure tumor necrosis factor-α (TNF-α), interleukin-8 (IL-8), malondialdehyde, protein carbonyl, IgA, microbiota in jejunal and colonic mucosa, morphology and crypt cell proliferation rate in the jejunum. Data were analyzed using the MIXED procedure of SAS. Sows fed LPL tended to increase (P = 0.084) litter size (11.9 vs. 12.6) on d 18 of lactation and decrease (P = 0.079) ADFI (8.72 vs. 8.02 kg) during d 9 to d 18 of lactation. Sows fed LPL tended to increase (P = 0.092) IgG (1.14 vs. 1.94 g/L) in the milk. Sows fed LPL increased (P < 0.05) crypt cell proliferation rate (39.38 vs. 40.94%) in the jejunum. Supplementation of lysophospholipids in lactation diet did not affect proinflammatory cytokines, oxidative stress markers, and microbiota in jejunum and colon of piglets on d 18 of lactation. In conclusion, supplementation of dietary lysophopholipids improved productive performance and the intestinal cell proliferation of piglets with enhancing IgG concentration in the milk.

Supplemental effects of dietary lysophospholipids in lactation diets on sow performance, milk composition, gut health, and gut-associated microbiome of offspring

Dietary lysophospholipids (LPL) would influence milk composition of sows, thus positively affect intestinal health of offspring. The objective of this study was to determine effects of dietary LPL fed to lactating sows on performance, milk characteristics, gut health, and gut-associated microbiome of offspring. Sixty pregnant sows were allotted to 2 treatments in a randomized complete block design with parity and BW as blocks on day 110 of gestation. Treatments were CON (no added LPL) and LPL (0.05% LPL; Lipidol-Ultra, Pathway Intermediates, Shrewsbury, UK). Sows were fed 2 kg/d from day 110 of gestation until farrowing and ad libitum after farrowing. Diets were formulated to meet NRC requirement for lactating sows. Colostrum and milk samples from 12 sows per treatment were collected to measure nutrients and immunoglobulins on days 1 and 18 of lactation, respectively. Twelve piglets per treatment (1 piglet per litter) were euthanized on day 18 to collect tissues to measure tumor necrosis factor-α, interleukin-8 (IL-8), malondialdehyde, protein carbonyl, IgA, histomorphology, crypt cell proliferation rate, and microbiota in the jejunum and colon. Data were analyzed using the MIXED procedure of SAS, and the mortality was analyzed using the GLIMMIX procedure of SAS. There was no difference in sow BW, parity, and litter size between treatments on day 0 of lactation. Sows fed LPL had increased (P < 0.05) litter BW gain (53.9 vs. 59.4 kg) and decreased piglet mortality (13.9% vs. 10.6%) on day 18 of lactation. Sows fed LPL had increased (P < 0.05) omega-6:omega-3 (22.1 vs. 23.7) and unsaturated:saturated (1.4 vs. 1.6) fatty acids ratios with increased oleic acid (29.1% vs. 31.4%) and tended to have increased (P = 0.092) IgG (1.14 vs. 1.94 g/L) and linoleic acid (17.7% vs. 18.7%) in the milk on day 18 of lactation. Piglets from sows fed LPL had increased (P < 0.05) IL-8 (184 vs. 245 pg/mg) and crypt cell proliferation rate (39.4% vs. 40.9%) and tended to have increased (P = 0.095) Firmicutes:Bacteroidetes ratio (1.0 vs. 3.5) in the jejunum. In conclusion, sows fed with LPL had milk with increased IgG, oleic acids, and linoleic acids without changes in BW and backfat during lactation. These changes could contribute to improved survivability and intestinal health of piglets by increasing IL-8 concentration, enhancing balance among gut-associated microbiome, and increasing enterocyte proliferation in the jejunum.

GLP-2, EGF, and the Intestinal Epithelial IGF-1 Receptor Interactions in the Regulation of Crypt Cell Proliferation

Glucagon-like peptide-2 (GLP-2) is an intestinotrophic hormone that promotes intestinal growth and proliferation through downstream mediators, including epidermal growth factor (EGF) and insulin-like growth factor-1 (IGF-1). EGF synergistically enhances the proliferative actions of IGF-1 in intestinal cell lines, and both of these factors are known to be essential for the trophic effects of GLP-2 in vivo. However, whether EGF and IGF-1 interact to mediate the proliferative actions of GLP-2 in vivo remains unknown. Normal and knockout (KO) mice lacking the intestinal epithelial IGF-1 receptor (IE-IGF-1R) were therefore treated chronically with EGF and/or long-acting human hGly2GLP-2, followed by determination of intestinal growth parameters. Intestines from control and IE-IGF-1R KO mice were also used to generate organoids (which lack the GLP-2 receptor) and were treated with EGF and/or IGF-1. Combination treatment with EGF and hGly2GLP-2 increased small intestinal weight and crypt-villus height in C57Bl/6 mice in an additive manner, whereas only hGly2GLP-2 treatment increased crypt cell proliferation. However, although combination treatment also increased small intestinal weight and crypt-villus height in IE-IGF-1R KO mice, the proliferative responses to hGly2GLP-2 alone or with EGF were diminished in these animals. Finally, IGF-1 treatment of organoids undergoing EGF withdrawal was not additive to the effect of EGF replacement on proliferation, but could restore normal proliferation in the absence of EGF. Together, these findings demonstrate that the intestinal proliferative effects of hGly2GLP-2 are augmented by exogenous EGF in a manner that is partially dependent upon IE-IGF-1R signaling.

Supplemental effects of dietary nucleotides on intestinal health and growth performance of newly weaned pigs

Intestinal challenges upon weaning would increase the needs of nucleotides for enterocyte proliferation, whereas de novo synthesis maybe insufficient. This study aimed to evaluate supplemental effects of dietary nucleotides on intestinal health and growth performance in newly weaned pigs. Fifty newly weaned pigs (19-d-old, 25 barrows and 25 gilts, 4.76 ± 0.42 kg BW) were individually housed and allotted to 5 treatments with increasing nucleotide supplementation (0, 50, 150, 250, and 500 mg/kg) based on a randomized complete block design with the initial BW and sex as blocks. Dietary nucleotides were provided from YT500 (Hinabiotech, Guangzhou, China). Pigs were fed for 21 d based on 2 phases (phase 1: 11 d and phase 2: 10 d) and experimental diets were formulated to meet or exceed nutrient requirements suggested by NRC (2012). Feed intake and BW were recorded. Titanium oxide (0.4%) was added as an indigestible marker from day 17. Plasma collected on day 18 was used to measure tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and malondialdehyde (MDA). Pigs were euthanized on day 21 to collect tissues to evaluate TNF-α, IL-6, MDA, morphology, and crypt cell proliferation rate in the jejunum. Ileal digesta were collected to measure ileal nutrient digestibility. Data were analyzed using contrasts in the MIXED procedure of SAS. Nucleotide supplementation increased (P < 0.05) ADFI in phase 1. Nucleotide supplementation at 50 and 150 mg/kg increased (P < 0.05) ADG in phase 1, whereas increased (P < 0.05) ADFI and tended to increase (P = 0.082) ADG in overall. Increasing nucleotide supplementation changed (quadratic, P < 0.05) villus height-crypt ratio (at 247 mg/kg) and decreased (linear, P < 0.05) crypt cell proliferation rate in the jejunum. Increasing nucleotide supplementation reduced (P < 0.05) jejunal IL-6 (at 50 and 150 mg/kg) and tended to change (quadratic, P = 0.074) plasma MDA (at 231 mg/kg). Nucleotide supplementation at 50 and 150 mg/kg increased (P < 0.05) ileal digestibility of energy and ether extract. In conclusion, nucleotide supplementation at a range of 50 to 250 mg/kg in the diets seems to be beneficial to newly weaned pigs by enhancing growth performance possibly due to reduced intestinal inflammation and oxidative stress as well as improved intestinal villi structure and energy digestibility.