Objective. To evaluate the in vivo therapeutic effect of pioglitazone, a peroxisome proliferatoractivated receptor ␥ (PPAR␥) agonist, on the development of lesions in a guinea pig model of osteoarthritis (OA), and to determine the influence of pioglitazone on the synthesis of matrix metalloproteinase 13 (MMP-13) and interleukin-1␤ (IL-1␤) in articular cartilage.Methods. The OA model was created by partial medial meniscectomy of the right knee joint. The guinea pigs were divided into 4 treatment groups: unoperated animals that received no treatment (normal), operated animals (OA guinea pigs) that received placebo, OA guinea pigs that received oral pioglitazone at 2 mg/kg/ day, and OA guinea pigs that received oral pioglitazone at 20 mg/kg/day. The animals began receiving medication 1 day after surgery and were killed 4 weeks later. Macroscopic and histologic analyses were performed on the cartilage. The levels of MMP-13 and IL-1␤ in OA cartilage chondrocytes were evaluated by immunohistochemistry.Results. OA guinea pigs treated with the highest dosages of pioglitazone showed a significant decrease, compared with the OA placebo group, in the surface area (size) and grade (depth) of cartilage macroscopic lesions on the tibial plateaus. The histologic severity of cartilage lesions was also reduced. A significantly higher percentage of chondrocytes in the middle and deep layers stained positive for MMP-13 and IL-1␤ in cartilage from placebo-treated OA guinea pigs compared with normal controls. Guinea pigs treated with the highest dosage of pioglitazone demonstrated a significant reduction in the levels of both MMP-13 and IL-1␤ in OA cartilage.Conclusion. This is the first in vivo study demonstrating that a PPAR␥ agonist, pioglitazone, could reduce the severity of experimental OA. This effect was associated with a reduction in the levels of MMP-13 and IL-1␤, which are known to play an important role in the pathophysiology of OA lesions.Osteoarthritis (OA) is a degenerative disease and the major cause of disability in humans. Aging, mechanical stress and traumatic injury, genetic susceptibility, and metabolic predispositions are considered risk factors for this disease. An important feature of OA is the degradation of articular cartilage, composed of abundant extracellular matrix rich in sulfated proteoglycan and type II collagen. This process is likely related to the excess synthesis and release of several catabolic factors such as proinflammatory cytokines, matrix metalloproteinases (MMPs), and nitric oxide (NO) in the tissue (1).OA is characterized by a shift of the balance between production of cartilage matrix and proteolytic degradation toward increased proteolysis. The most influential causative proteases in OA are MMPs, a family of zinc-containing, calcium-dependent proteases. An MMP that is of particular interest in the degradation of cartilage in pathologic conditions is MMP-13 (collagenase 3). MMP-13 has been found to be elevated in both rheumatoid arthritis (RA) and OA joint tissues and, more particularly, ...

Matrix metalloproteinase-13 (MMP-13) has been implicated to play a key role in the pathology of osteoarthritis. On the basis of X-ray crystallography, we designed a series of potent MMP-13 selective inhibitors optimized to occupy the distinct deep S1' pocket including an adjacent branch. Among them, carboxylic acid inhibitor 21k exhibited excellent potency and selectivity for MMP-13 over other MMPs. An effort to convert compound 21k to the mono sodium salt 38 was promising in all animal species studied. Moreover, no overt toxicity was observed in a preliminary repeat dose oral toxicity study of compound 21k in rats. A single oral dose of compound 38 significantly reduced degradation products (CTX-II) released from articular cartilage into the joint cavity in a rat MIA model in vivo. In this article, we report the discovery of highly potent, selective, and orally bioavailable MMP-13 inhibitors as well as their detailed structure-activity data.

Background:The in vivo mechanism of autophagy regulation remains unclear. Results: The effect of insulin on mTORC1-autophagy is greater in muscle than in the liver, whereas that of amino acids is greater in liver than in muscle. Conclusion: Autophagy is differentially regulated by insulin and amino acids in a tissue-dependent manner. Significance: This information increases the understanding of protein homeostasis in vivo.

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On the basis of a superposition study of X-ray crystal structures of complexes of quinazoline derivative 1 and triazole derivative 2 with matrix metalloproteinase (MMP)-13 catalytic domain, a novel series of fused pyrimidine compounds which possess a 1,2,4-triazol-3-yl group as a zinc binding group (ZBG) was designed. Among the herein described and evaluated compounds, 31f exhibited excellent potency for MMP-13 (IC = 0.036 nM) and selectivities (greater than 1,500-fold) over other MMPs (MMP-1, -2, -3, -7, -8, -9, -10, and -14) and tumor necrosis factor-α converting enzyme (TACE). Furthermore, the inhibitor was shown to protect bovine nasal cartilage explants against degradation induced by interleukin-1 and oncostatin M. In this article, we report the discovery of extremely potent, highly selective, and orally bioavailable fused pyrimidine derivatives that possess a 1,2,4-triazol-3-yl group as a novel ZBG for selective MMP-13 inhibition.

Polycomb-group (PcG) proteins play crucial roles in self-renewal of stem cells by suppressing a host of genes through histone modifications. Identification of the downstream genes of PcG proteins is essential for elucidation of the molecular mechanisms of stem cell self-renewal. However, little is known about the PcG target genes in tissue stem cells. We found that the PcG protein, Ring1B, which regulates expression of various genes through monoubiquitination of histone H2AK119, is essential for expansion of hepatic stem/progenitor cells. In mouse embryos with a conditional knockout of Ring1B, we found that the lack of Ring1B inhibited proliferation and differentiation of hepatic stem/progenitor cells and thereby inhibited hepatic organogenesis. These events were characterized by derepression of cyclin-dependent kinase inhibitors (CDKIs) Cdkn1a and Cdkn2a, known negative regulators of cell proliferation. We conducted clonal culture experiments with hepatic stem/progenitor cells to investigate the individual genetic functions of Ring1B, Cdkn1a, and Cdkn2a. The data showed that the cell-cycle inhibition caused by Ring1B depletion was reversed when Cdkn1a and Cdkn2a were suppressed simultaneously, but not when they were suppressed individually. Conclusion: Our results show that expansion of hepatic stem/progenitor cells requires Ring1B-mediated epigenetic silencing of Cdkn1a and Cdkn2a, demonstrating that Ring1B simultaneously regulates multiple CDKIs in tissue stem/progenitor cells. (HEPATOLOGY 2014;60:323-333)

Pulmonary hypertension (PH) is a life-threatening lung disease. PH with concomitant lung diseases, e.g., idiopathic pulmonary fibrosis, is associated with poor prognosis. Development of novel therapeutic vasodilators for treatment of these patients is a key imperative. We evaluated the efficacy of dual activation of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) using an active, small-molecule phosphodiesterase (PDE4)/PDE5 dual inhibitor (Compound A). Compound A increased both cAMP and cGMP levels in WI-38 lung fibroblasts and suppressed the expressions of type-1 collagen α1 chain and fibronectin. Additionally, compound A reduced right ventricular weight/left ventricular weight+septal weight ratio, brain natriuretic peptide expression levels in right ventricle, C─C motif chemokine ligand 2 expression levels in lung, and plasma surfactant protein D. Our data indicate that dual activation of cAMP/cGMP pathways may be a novel treatment strategy for PH.

Role of prostaglandins on feeding-associated induction of ornithine decarboxylase in small intestine was studied. Rats received intraperitoneal injection of either saline, or 16,16-dimethyl prostaglandin E2, or TRY-200 (a stable prostaglandin I2 analog), or refeeding, after a 44 hr-fast. Four hours later, mucosae from duodenum, jejunum, and ileum were scraped for subsequent measurements of enzyme activity of ornithine decarboxylase by a radiometric technique. Refeeding resulted in a profound induction of enzyme activity throughout the small intestine. Parenteral administration of prostaglandin I2 also led to a significant induction with the level similar to refeeding. The stimulatory effect of prostaglandin I2 was completely abolished by a specific and irreversible enzyme inhibitor, difluoromethylornithine. Prostaglandin E2 had a similar but lesser effect than prostaglandin I2 on the induction of the enzyme activity. Pretreatment with indomethacin, a cyclooxygenase inhibitor had no effect on feeding-associated enzyme induction. These results indicate that although exogenous prostaglandin I2 appears to be a potent stimulant for ornithine decarboxylase activity in rat small intestine, endogenous prostaglandins seem to play little or no role in feeding-associated induction of ornithine decarboxylase.