LE 135

All-trans-retinoic Acid Increases SLC26A3 DRA (Down-regulated in Adenoma) Expression in Intestinal Epithelial Cells via HNF-1beta.[Pubmed:25887398]

J Biol Chem. 2015 Jun 12;290(24):15066-77.

All-trans-retinoic acid (ATRA) is an active vitamin A derivative known to modulate a number of physiological processes, including growth and development, differentiation, and gene transcription. The protective effect of ATRA in gut inflammation and diarrheal diseases has been documented. In this regard, down-regulated in adenoma (DRA, a key luminal membrane Cl(-) transporter involved in NaCl absorption) has been shown to be suppressed in intestinal inflammation. This suppression of DRA is associated with diarrheal phenotype. Therefore, current studies were undertaken to examine the effects of ATRA on DRA expression. DRA mRNA levels were significantly elevated ( approximately 4-fold) in response to ATRA with induction starting as early as 8 h of incubation. Similarly, ATRA increased DRA protein expression by approximately 50%. Furthermore, DRA promoter activity was significantly increased in response to ATRA indicating transcriptional activation. ATRA effects on DRA expression appeared to be mediated via the RAR-beta receptor subtype, as ATRA remarkably induced RAR-beta mRNA levels, whereas RAR-beta knockdown substantially attenuated the ability of ATRA to increase DRA expression. Results obtained from agonist (CH-55) and antagonist (LE-135) studies further confirmed that ATRA exerts its effects through RAR-beta. Furthermore, ATRA treatment resulted in a significant increase in HNF-1beta mRNA levels. The ability of ATRA to induce DRA expression was inhibited in the presence of HNF-1beta siRNA indicative of its involvement in ATRA-induced effects on DRA expression. In conclusion, ATRA may act as an antidiarrheal agent by increasing DRA expression via the RAR-beta/HNF-1beta-dependent pathway.

Chondrogenic effects of exogenous retinoic acid or a retinoic acid receptor antagonist (LE135) on equine chondrocytes and bone marrow-derived mesenchymal stem cells in monolayer culture.[Pubmed:21728848]

Am J Vet Res. 2011 Jul;72(7):884-92.

OBJECTIVE: To determine effects of various concentrations of retinoic acid (RA) or a synthetic RA receptor antagonist (LE135) on equine chondrocytes or bone marrow-derived equine mesenchymal stem cells (BMDMSCs) in monolayer cultures. SAMPLE: Articular cartilage and BMDMSCs from 5 clinically normal horses. PROCEDURES: Monolayers of chondrocytes cultured in standard media and of BMDMSCs cultured in chondrogenic media were treated with RA at concentrations of 0, 0.1, 1, or 10 muM or LE135 at concentrations of 0, 0.1, 1, or 10 muM on day 0. On days 7 and 14, samples were analyzed for DNA concentration, chondrocyte morphology or features consistent with chondrogenesis (ie, chondral morphology [scored from 0 to 4]), and gene expression of collagen type Ia (CI), collagen type II (CII), and aggrecan. RESULTS: Chondrocytes treated with RA had more mature chondral morphology (range of median scores, 3.0 to 4.0) than did untreated controls (range of median scores, 0.5 to 0.5). Chondrocytes treated with LE135 did not sustain chondrocyte morphology. All BMDMSCs had evidence of chondral morphology or high CII:CI ratio. Retinoic acid (1 or 10 muM) or LE135 (10 muM) treatment decreased DNA content of BMDMSC cultures. At 0.1 and 1 muM concentrations, LE135 weakly but significantly increased chondral morphology scores, compared with untreated controls, but lack of aggrecan expression and lack of increased CII:CI ratio, compared with that of controls, did not affect chondrogenesis. CONCLUSIONS AND CLINICAL RELEVANCE: RA promoted maturation and hypertrophy in chondrocytes but not BMDMSCs in monolayer cultures. Deficiency or blockade of RA may prevent hypertrophy and maturation of differentiated chondrocytes.

All-trans retinoic acid ameliorates trinitrobenzene sulfonic acid-induced colitis by shifting Th1 to Th2 profile.[Pubmed:20187766]

J Interferon Cytokine Res. 2010 Jun;30(6):399-406.

Inflammatory bowel disease is characterized with uncontrolled immune response in inflamed mucosa, with dominance of Th1 cells. Recently, all-trans retinoic acid has been shown that can lead T-cell response by suppressing Th17 development via retinoic acid receptor (RAR), but it is still unknown whether all-trans retinoic acid can modulate Th1 response of inflammatory bowel disease. In the experiment, we investigated the effect of all-trans retinoic acid on trinitrobenzene sulfonic acid (TNBS)-induced murine colitis, and the possible mechanism. Mice were intraperitoneally treated daily with all-trans retinoic acid (the agonist of RAR-alpha) or LE135 (the antagonist of RAR-alpha) or medium, and sacrificed 6 days later. Colon was collected for histological analysis and myeloperoxidase (MPO) activity measurement. Lamina propria mononuclear cells (LPMCs) were isolated, cultured, and assayed for the expressions of T-bet and GATA-3 by the use of Western blot and for cytokine levels by the use of ELISA. All-trans retinoic acid treatment inhibited inflammatory responses as shown by lower histological inflammatory scores and MPO activity, compared with LE135 and medium groups. Furthermore, in LPMCs culture supernatants, the levels of Th1 cytokines (INF-gamma, IL-12, and TNF-alpha) were decreased while those of Th2 cytokines (IL-4 and IL-10) were increased significantly in all-trans retinoic acid-treated mice. In addition, T-bet expression in LPMCs was inhibited and GATA-3 expression was up-regulated in all-trans retinoic acidtreated mice. On the contrary, LE135 showed the reverse effects in colon inflammation and cytokine profile. By shifting Th1 to Th2 profile in inflamed mucosa, all-trans retinoic acid down-regulates inflammatory response and ameliorates acute TNBS-induced colitis, which suggests the ligand of RAR-alpha-based pharmaceutical strategies for managing inflammatory bowel disease.

Anti-hyperglycemic effect of Opuntia streptacantha Lem.[Pubmed:21111796]

J Ethnopharmacol. 2011 Jan 27;133(2):940-3.

AIM OF THE STUDY: This experiment studied two extracts of Opuntia streptacantha, a plant used by the Mexican population to treat type 2 diabetes, in different assays to contribute to the understanding of the hypoglycemic mechanism of this plant. MATERIALS AND METHODS: Two different extracts were prepared and tested: the first extract was a filtrate of the traditional liquefied extract (LE) preparation of the cladode; and the second filtrate extract (FE) is a filtered sample of the first. Both extracts contained a newly identified compound for Opuntia (4-hydroxy)-phenyl acetic acid derivate, they were tested on streptozotocin (STZ)-diabetic rats in a series of two tests. The first test was performed to confirm if STZ-diabetic rats presented a hypoglycemic effect after administration of the extracts (LE 135 mg/kg and FE 27 mg/kg). In the second experiment, the extracts were administered before an oral glucose tolerance test (OGTT) to confirm if they have an anti-hyperglycemic effect (LE 135 mg/kg, FE 12 and 27 mg/kg). RESULTS: The extracts administered to STZ-diabetic rats did not produce a significant hypoglycemic effect compared to the control group, while the same extracts administered before an OGTT produced an anti-hyperglycemic effect compared to the control group. CONCLUSIONS: The filtered, traditional LE of the cladode of Opuntia streptacantha produces an anti-hyperglycemic effect when administered before a glucose challenge, and this anti-hyperglycemic effect is maintained after filtering the extract. Administration of both plants can improve glycemic control by blocking the hepatic glucose output, especially in the fasting state. These data support the traditional use of the plants as "agua de uso", a cold infusion of the plant consumed over the course of a day.

All-trans retinoic acid down-regulates inflammatory responses by shifting the Treg/Th17 profile in human ulcerative and murine colitis.[Pubmed:19477911]

J Leukoc Biol. 2009 Oct;86(4):959-69.

IBD is characterized by uncontrolled immune responses in inflamed mucosa, with dominance of IL-17-producing cells and deficiency of Treg cells. The aim of this study was to explore the effect and mechanisms of RA, the ligand of RARalpha, on immune responses in human and murine colitis. Colonic biopsies from patients with UC were cultured and treated with RA as the agonist of RARalpha or LE135 as the antagonist of RARalpha. Expressions of IL-17 and FOXP3 were detected by immunohistochemistry. Murine colitis was induced by intrarectal administration with TNBS at Day 1. Mice were then i.p.-treated with RA or LE135 daily for 7 days. Cytokine levels in the cultures of mouse LPMCs were measured. Expressions of FOXP3 and IL-17 in colon tissues or MLN were detected by immunohistological analysis. Body weight and colon inflammation were evaluated. RA treatment up-regulated FOXP3 expression and down-regulated IL-17 expression in colon biopsies of patients and in colon tissues and MLN of mice with colitis compared with controls. LPMCs from RA-treated mice produced lower levels of proinflammatory cytokines (TNF-alpha, IL-1beta, IL-17) but more regulatory cytokines (IL-10, TGF-beta) compared with that of untreated mice. LE135 showed the opposite effect of RA. Furthermore, RA ameliorated TNBS-induced colitis in a dose-dependent manner, as seen by improved body weight and colon inflammation. RA down-regulates colon inflammatory responses in patients with IBD in vitro and in murine colitis in vivo, representing a potential therapeutic approach in IBD treatment.