Correlation between Plasma Glucagon-Like Peptide 2 Levels and Proliferative Makers in Small Intestinal Injury in Rats Induced by Methotrexate Administration

Abstract

Chemotherapy is an important primary and adjuvant therapy for cancer patients. The cytotoxicity of antineoplastic agents affects not only tumor cells but also rapidly proliferating normal cells, such as those of the gastrointestinal mucosa.1) Common complications of chemotherapy thus include stomatitis and enterocolitis.2) Methotrexate (MTX) is an antimetabolite drug that blocks the production of biologically active forms of folic acid. The major lesions resulting from its cytotoxic effects occur in bone marrow and the intestinal tract. High doses of MTX cause severe enterocolitis, weight loss, anorexia and intestinal atrophy. 3,4) The mucosa of the digestive tract, particularly that of the small intestine, is a critical site of action of anticancer agents including MTX, as the undifferentiated epithelial cells in the crypts, which ensure continuous and relatively fast regeneration of the epithelium, are particularly sensitive to these cytostatic drugs.5-7) Patients receiving these anticancer agents frequently experience gastrointestinal side effects, such as nausea, vomiting and diarrhea, reflecting gastrointestinal mucosal damage or enterocolitis.2) Chemotherapy-associated diarrhea may be problematic, and even life threatening, if complicated by mucosal ulceration and inflammatory changes. Moreover, patients with advanced malignancy often suffer from malnutrition, which increases toxicity and decreases tumor responses to treatment,2) and such conditions can be dose limiting in therapeutic regimens. A method for prediction of adverse reactions to chemotherapy is thus needed to avoid their occurrence.Gastrointestinal mucosal integrity is regulated by several growth factors and cytokines, such as transforming growth factors-beta and IL-11.8-11) Glucagon-like peptide 2 (GLP-2) has been demonstrated to have intestinotrophic effects. 12)GLP-2, a 33-amino acid peptide member of a family of proglucagon-derived peptides (PGDP) that also includes enteroglucagon, is secreted from L-type entero-endocrine cells of the distal small intestine and colon.12) GLP-2 increases crypt cell proliferation and decreases the rate of apoptosis in small intestinal epithelium.13) Acute or chronic administration of GLP-2 induces functional changes in the intestine, improving its capacity for nutrient absorption. [13][14][15] In addition, GLP-2 treatment has been shown to enhance intestinal adaptation following major small bowel resection, 16,17) to prevent 18) and to reduce the severity of dextran sulfate-induced colitis 19) and indomethacin-induced enteritis 20) in animal models. On the other hand, plasma GLP-2 levels appear to vary with intestinal mucosal condition and injury. However, although changes in various conditions of the intestinal mucosa are now being widely studied, [21][22][23] the relationship between plasma GLP-2 levels and intestinal mucosal injury induced by various drugs, including antineoplastic agents, has yet to be clearly determined. Moreover, no standard methods of diagnosis or therapeutic protocols exist for chemo...

The prolonged effect of glucagon-like peptide 2 pretreatment on growth performance and intestinal development of weaned piglets

“…These findings reveal a relationship between the GLP-2 level and intestinal adaptation. Previous studies showed that supplementation of enteral nutrients or exogenous GLP-2 induced an increase in plasma concentration of GLP-2 in neonatal piglets and rats with intestinal injury or short bowel syndrome [ 3 , 4 , 12 ]. Similarly, in the present study, injection of a high dose of GLP-2 significantly prevented the decline of basal levels of plasma GLP-2 and increased the GLP-2 above the weaning level.…”

Sodium Alginate Inhibits Methotrexate-Induced Gastrointestinal Mucositis in Rats

“…14,17) In rats, methotrexate-induced gastrointestinal mucositis is characterized histologically by crypt loss, callus fusion and atrophy, capillary dilatation, and infiltration of mixed inflammatory cells. 15,18,19) These side effects necessitate dose reduction and discontinuation of selected cancer therapies, and increase healthcare costs, prolong hospital stays, increase re-admission rates, compromise patient nutritional status, impair patient quality of life, and are occasionally fatal. Thus, therapeutic agents that ameliorate chemotherapy-induced gastrointestinal mucositis are urgently required.…”

Potentiation of Glucagon-Like Peptide-2 Dynamics by Methotrexate Administration in Rat Small Intestine

“…13) RNA Extraction and RT-PCR Messenger RNA expression was quantified by real-time RT-PCR using a 7500 Fast Real-Time PCR system (Life Technologies, Applied Biosystems, Foster City, CA, U.S.A.) and PrimeScript One Step RT-PCR kit (TaKaRa Bio, Shiga, Japan), as previously described. 14) The following sense and antisense primers, respectively, were used: proglucagon, 5′-AAC AAC ATT GCC AAA CGT CA-3′ and 5′-CAA TGA ATT CCT TTG CTG CC-3′; GLP-2 receptor, 5′-CGG ATT CTG GAA ATT CTT C-3′ and 5′-CCC CAG GAA CCG GAA ATT CTT C-3′; IGF-1, 5′-AAG CCT ACA AAG TCA GCT CG-3′ and 5′-GGT CTT GTT TCC TGC ACT TC-3′; DPP-4, 5′-TCC CAA CTC CAG AGG ACA AC-3′ and 5′-CAG GGT TTG GAG TCT GAG-3′; TGF-β1, 5′-TGG CGT TAC CTT GGT AAC C-3′ and 5′-CGT GTT GAG CCC TTT CCA G-3′; TGF-β2, 5′-ATC GAT GGC ACC TCC ACA TAT G-3′ and 5′-GCG AAG GCA GCA ATT ATG CTG-3′; TGF-β3, 5′-AAG CGC ACA GAG CAG AGA ATC-3′ and 5′-AGT GTC AGT GAC ATC GAA G 3′; β-catenin, 5′-GCC AAG TGG GTG GCA TAG A-3′ and 5′-TCC CTG TCA CCA GCA CGA A-3′; the exchange protein activated by cAMP 1 (Epac1), 5′-GTG TTG GTG AAG GTC AAT TCT G-3′ and 5′-CCA CAC CAC GGG CAT C-3′; Epac2, 5′-TGT TAA AGT GTC TGA GAC CAG CA-3′ and 5′-AAA GGC TGT CCC AAT TCC CAG-3′; and glyceraldehyde 3-phosphate dehydrogenase, 5′-ATG TTC CAG TAT GAC TCC ACT CAC G-3′ and 5′-GAA GAC ACC AGT AGA CTC CAC GAC A-3′. The PCR products were calculated relative to glyceraldehyde 3-phosphate dehydrogenase.…”