sal repair and enhanced enterocyte turnover. While the proliferative zone in MTX-rats moved progressively upwards in the crypts toward the crypt-villus junction, the proliferative zone of MTX- TGFb2 rats was only mildly affected, showing a slight shift upwards within the crypts. In addition, exposure to oral TGFb2 significantly enhanced intestinal recovery following MTXinduced damage. This is evident from the significant increase in bowel and mucosal weight, increased DNA and protein content in ileum. Histologically, marked increases in villus height suggest increased absorptive surface area and closely correlate with increased cell mass. Similar to control rats, TGFb2 supplementation resulted in a significant increase in mucosal cell proliferation in functioning MedChemExpress BIX01294 intestine, but decreased significantly cell apoptosis PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/22210242 rate, which may represent the main mechanism that maintains mucosal structure following MTX-induced damage. Enhanced cell proliferation in the current study was correlated with elevated bcatenin and p-ERK protein levels that may suggest an activated stem cell activity and stimulated MAPK signaling pathway. Our results show also that the intrinsic pathway, with its regulation by the bcl-2 family of proteins, was altered by TGFb2 in accordance with changes in cell apoptosis: the mRNA and protein levels of the pro-apoptotic bax decreased, while those of the antiapoptotic bcl2 mRNA levels increased. Correspondingly, bax/bcl-2 ratio decreased in MTX- TGFb rats compared to MTX animals, suggesting increased enterocyte survival. Further investigation is needed to define the regulation of this special apoptotic state with respect to the Fas/Fasl-mediated extrinsic pathway. This positive effect was accompanied by decreased levels of IL-1B protein in intestinal mucosa, suggesting anti-inflammatory effect of TGF- b2. Next, we investigated whether the effects of TGF-b2 on enterocyte proliferation and apoptosis were correlated with TGF-b2 receptor expression throughout the gastrointestinal tract and along the villuscrypt axis. We have shown that the pro-proliferative and anti-apoptotic effect of TGF-b2 on enterocyte turnover is correlated with elevated TGF-b2 receptor expression following TGF-b2 administration. In the crypt compartment, a significant increase in TGFb2 receptor expression following TGF-b2 administration coincided with increased cell proliferation. In villus tips, MTX- TGF-b rats demonstrated higher receptor immunoreactivity compared to MTX animals. Since TGF-b exerts anti-apoptotic effects, this increase in TGF-b2 receptor expression coincides with decreased cell apoptosis in villus tips following TGF-b2 administration. In conclusion, treatment with TGF-b2 
increased cell viability and decreased of cell apoptosis in Caco-2 cell line. In a rat model of MTX-induced mucositis, dietary TGF-b2 supplementation reverse intestinal damage, causes anti-inflammatory effect and stimulates intestinal recovery. Enhanced cell proliferation and inhibited programmed cell death may be responsible for this effect. The pro-proliferative and anti-apoptotic effects of TGF-b2 are correlated with TGF-b2 receptor expression along the villus-crypt axis. Dietary TGF-b2 may be clinically beneficial as an agent to prevent intestinal damage and stimulate intestinal recovery in patients with chemotherapy-induced mucositis. TGF-b2 Reduces MTX Induced Intestinal Injury Clathrin adaptor protein complexes play a key role in the transport of proteins by regul
