Dietary Transforming Growth Factor-Beta 2 (TGF-β2) Supplementation Reduces Methotrexate-Induced Intestinal Mucosal Injury in a Rat

Abstract

Background/AimsDietary supplementation with transforming growth factor-beta (TGF-β) has been proven to minimize intestinal damage and facilitate regeneration after mucosal injury. In the present study, we evaluated the effects of oral TGF-β2 supplementation on intestinal structural changes, enterocyte proliferation and apoptosis following methotrexate (MTX)-induced intestinal damage in a rat and in a cell culture model.MethodsCaco-2 cells were treated with MTX and were incubated with increasing concentrations of TGF-β2. Cell apoptosis was assessed using FACS analysis by annexin staining and cell viability was monitored using Trypan Blue assay. Male rats were divided into four experimental groups: Control rats, CONTR- TGF-β rats were treated with diet enriched with TGF-β2, MTX rats were treated with a single dose of methotrexate, and MTX- TGF-β rats were treated with diet enriched with TGF-β2. Intestinal mucosal damage, mucosal structural changes, enterocyte proliferation and enterocyte apoptosis were determined at sacrifice. Real Time PCR and Western blot were used to determine bax and bcl-2 mRNA, p-ERK, β-catenin, IL-1B and bax protein expression.ResultsTreatment of MTX-pretreated Caco-2 cells with TGF-B2 resulted in increased cell viability and decreased cell apoptosis. Treatment of MTX-rats with TGF-β2 resulted in a significant increase in bowel and mucosal weight, DNA and protein content, villus-height (ileum), crypt-depth (jejunum), decreased intestinal-injury score, decreased level of apoptosis and increased cell proliferation in jejunum and ileum compared to the untreated MTX group. MTX-TGF-β2 rats demonstrated a lower bax mRNA and protein levels as well as increased bcl-2 mRNA levels in jejunum and ileum compared to MTX group. Treatment with TGF-β2 also led to increased pERK, IL-1B and β-catenin protein levels in intestinal mucosa.ConclusionsTreatment with TGF-β2 prevents mucosal-injury, enhances p-ERK and β-catenin induced enterocyte proliferation, inhibits enterocyte apoptosis and improves intestinal recovery following MTX-induced intestinal-mucositis in rats.

Lactobacillus rhamnosus GG on Rotavirus‐Induced Injury of Ileal Epithelium in Gnotobiotic Pigs

“…By PID 2, the shortened villi with compact enterocytes and increased levels of villin, AJ and TJ proteins were evident in all the HRV inoculated Gn pigs. Study of methotrexate (MTX)-induced intestinal injury in rats reported the similar observation that β-catenin protein level increased in the treated rats compared to the controls at 72 hours after MTX-induced damage (43). More importantly, the decreased AJ and TJ protein expression in the ileum of LGG+HRV pigs compared to HRV alone pigs indicates that LGG attenuated the HRV-associated ileal injury and the associated compensatory/regenerative process after viral infection in the Gn pigs.…”

Bovine Colostrum Before or After Formula Feeding Improves Systemic Immune Protection and Gut Function in Newborn Preterm Pigs

“…In a previous study, we showed that formula-induced villus atrophy in preterm pigs may be due to increased enterocyte apoptosis (33), which is supported by the upregulation of an apoptosis gene, CASP3 in the formula-fed pigs in this study. Bioactive factors found in high amounts in colostrum, such as lactoferrin, TGF-β2, and IGF-I, have been shown to attenuate epithelial apoptosis in cell and animal models (34)(35)(36). In this study, colostrum feeding was able to prevent and restore formula-induced impairment in terms of villus atrophy, digestive and absorptive functions, and permeability, which may relate to its apoptosis-suppressed effects supported by the similarly downregulation of CASP3 in the CF and FC groups, relative to the FF group.…”

GWAS of 972 autologous stem cell recipients with multiple myeloma identifies 11 genetic variants associated with chemotherapy-induced oral mucositis

“…Fibrillin 2 encoded by FBN2 is expressed in keratinocytes [24], is one of the components of the extracellular matrix, and has been shown to play an important role in the regulation of the bioavailability of TGFβ in a tissue [25, 26]. Dietary supplementation of TGFβ-2 reduced methotrexate-induced intestinal mucositis in rats [27], but the effects of TGFβ-2 and TGFβ-3 administration on chemotherapy-induced mucositis in humans have yet to be demonstrated [28, 29]. In contrast, a recent study showed that Smad7 can reduce radiation-induced OM by dampening TGFβ signaling [30], It remains to be determined whether patients with a different SNP rsl0072361 exhibit a differential expression of fibrillin 2 and different TGFβ activity in correlation with their susceptibility to melphalan-induced OM.…”