Toxicity Of Nonsteroidal Anti-inflammatory Drugs Research Articles

Can NO-indomethacin counteract the topical gastric toxicity induced by indomethacin interactions with phospholipid bilayers?

Nitric oxide (NO)-releasing nonsteroidal anti-inflammatory drugs (NSAIDs) have been developed to overcome the gastrointestinal and cardiovascular toxicity of NSAIDs, by chemically associating a NO-releasing moiety with commercial NSAIDs. Since increasing evidence supports that NSAIDs toxicity is related to their topical actions in membrane lipids, this work aims to evaluate the impact of adding a NO-releasing moiety to parent NSAIDs regarding their effect on lipid bilayers. Thus, the interactions of NO-indomethacin and indomethacin (parent drug) with 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) bilayers were described herein at pH 3.0 and 7.4. Diverse experimental techniques were combined to characterize the partitioning and location of drugs in DMPC bilayers, and to analyze their effect on the lipid phase transition and the bilayer structure and dynamics. The partitioning of NO-indomethacin into DMPC bilayers was similar to that of charged indomethacin and smaller than that of neutral indomethacin. Both drugs were found to insert the DMPC bilayer and the membrane location of indomethacin was pH-dependent. NO-indomethacin and indomethacin induced a decrease of the main phase transition temperature of DMPC. The effect of these drugs on the bilayer structure and dynamics was dependent on diverse factors, namely drug ionization state, drug:lipid molar ratio, temperature and lipid phase. It is noteworthy that NO-indomethacin induced more pronounced alterations in the biophysical properties of DMPC bilayers than indomethacin, considering equivalent membrane concentrations. Such modifications may have in vivo implications, particularly in the gastric mucosa, where NO-NSAIDs-induced changes in the protective properties of phospholipid layers may contribute to the occurrence of adverse effects.

Discontinuing a non-steroidal anti-inflammatory drug (NSAID) in patients with knee osteoarthritis: Design and protocol of a placebo-controlled, noninferiority, randomized withdrawal trial

BackgroundKnee osteoarthritis (OA) is the most common cause of knee pain in older adults. Despite the limited data supporting their use, non-steroidal anti-inflammatory drugs (NSAID) are among the most commonly prescribed medications for knee OA. The use of NSAIDs for knee pain warrants careful examination because of toxicity associated with this class of medications. MethodsWe describe the design of a placebo-controlled, noninferiority, randomized withdrawal trial to examine discontinuation of an NSAID in patients with painful knee OA. Participants will be veterans enrolled in the VA Healthcare System with knee OA pain despite NSAID use and/or relatively higher risk of NSAID toxicity. After a two-week run-in period where eligible subjects will replace their current NSAID with the study NSAID (meloxicam), those remaining eligible (target N=544) will be randomized to receive four weeks of either placebo or continued meloxicam. The primary outcome is knee pain (Western Ontario and McMaster Universities Osteoarthritis Index pain subscale, range 0—20) at four weeks post-randomization. The primary hypothesis is that placebo will be noninferior to (that is, not much worse than) meloxicam within a noninferiority margin of 1. Secondary outcomes include lower extremity disability, global impression of change, adherence to study medication and use of co-therapies. DiscussionThis study is the first clinical trial to date examining the effects of withdrawing an NSAID for OA knee pain. If successful, this trial will provide evidence against the continued use of NSAIDs in patients with OA knee pain. Trial registrationClinicalTrials.gov: NCT01799213. Registered February 22, 2013.

Drug-Disease Interaction: Effect of Inflammation and Nonsteroidal Anti-Inflammatory Drugs on Cytochrome P450 Metabolites of Arachidonic Acid

Inflammatory conditions increase cardiovascular (CV) risk. Some nonsteroidal anti-inflammatory drugs (NSAIDs) that are used to treat pain and inflammation are also associated with CV complications. Inflammation, but not NSAIDs, disrupts the balance of vasodilator and vasoconstrictor components of the renin-angiotensin system within the heart. Herein, we report the effect of both inflammation and NSAIDs (rofecoxib, celecoxib, and meloxicam) on the physiologically active cytochrome P450 metabolites of arachidonic acid (ArA) in the rat with adjuvant arthritis. After oral administration of 7 daily therapeutically equivalent doses of NSAIDs or vehicle, the anti-inflammatory response, as well as the ArA metabolites and drug concentrations in plasma, heart and kidneys were assessed. Inflammation in the form of adjuvant arthritis caused a significant tissue-dependent imbalance of ArA metabolites by elevating the ratio of cardiotoxic 20-hydroxyeicosatetraenoic acid over cardioprotective epoxyeicosatrienoic acids in the heart, and reducing the ratio in the kidney. The observed imbalance was augmented by cardiotoxic rofecoxib but not by other examined NSAIDs with known milder cardiotoxicity. The cardio-renal toxicity of NSAIDs with known severe CV side effects may be due to altered cytochrome P450–mediated ArA acid metabolism. The ArA metabolism profile may be a marker of NSAIDs safety and toxicity.

The Risk of Major NSAID Toxicity with Celecoxib, Ibuprofen, or Naproxen: A Secondary Analysis of the PRECISION Trial

BackgroundThe relative safety of long-term use of nonsteroidal anti-inflammatory drugs is unclear. Patients and providers are interested in an integrated view of risk . We examined the risk of major nonsteroidal anti-inflammatory drug toxicity in the PRECISION trial. MethodsWe conducted a post hoc analysis of a double-blind, randomized, controlled, multicenter trial enrolling 24,081 patients with osteoarthritis or rheumatoid arthritis at moderate or high cardiovascular risk. Patients were randomized to receive celecoxib 100 to 200 mg twice daily, ibuprofen 600 to 800 mg thrice daily, or naproxen 375 to 500 mg twice daily. All patients were provided with a proton pump inhibitor. The outcome was major nonsteroidal anti-inflammatory drug toxicity, including time to first occurrence of major adverse cardiovascular events, important gastrointestinal events, renal events, and all-cause mortality. ResultsDuring follow-up, 4.1% of subjects sustained any major toxicity in the celecoxib arm, 4.8% in the naproxen arm, and 5.3% in the ibuprofen arm. Analyses adjusted for aspirin use and geographic region found that subjects in the naproxen arm had a 20% (95% CI 4-39) higher risk of major toxicity than celecoxib users and that 38% (95% CI 19-59) higher risk. These risks translate into numbers needed to harm of 135 (95% CI, 72-971) for naproxen and 82 (95% CI, 53-173) for ibuprofen, both compared with celecoxib. ConclusionsAmong patients with symptomatic arthritis who had moderate to high risk of cardiovascular events, approximately 1 in 20 experienced a major toxicity over 1 to 2 years. Patients using naproxen or ibuprofen experienced significantly higher risk of major toxicity than those using celecoxib.

REDUCED RISK OF NSAID INDUCED ADVERSE EVENTS WITH CONCOMITANT USE OF MISOPROSTOL

Background: Non-steroidal anti-inflammatory drugs (NSAIDs) are the most commonly used prescription and over-the-counter analgesic medications worldwide. NSAID toxicity, however, limits their use by raising cardio-renal morbidity and mortality risk. Withdrawal of the balancing vasodilatory

Nonsteroidal Anti-Inflammatory Therapy: A Journey Toward Safety.

The efficacy of nonsteroidal anti-inflammatory drugs (NSAIDs) against inflammation, pain, and fever has been supporting their worldwide use in the treatment of painful conditions and chronic inflammatory diseases until today. However, the long-term therapy with NSAIDs was soon associated with high incidences of adverse events in the gastrointestinal tract. Therefore, the search for novel drugs with improved safety has begun with COX-2 selective inhibitors (coxibs) being straightaway developed and commercialized. Nevertheless, the excitement has fast turned to disappointment when diverse coxibs were withdrawn from the market due to cardiovascular toxicity. Such events have once again triggered the emergence of different strategies to overcome NSAIDs toxicity. Here, an integrative review is provided to address the breakthroughs of two main approaches: (i) the association of NSAIDs with protective mediators and (ii) the design of novel compounds to target downstream and/or multiple enzymes of the arachidonic acid cascade. To date, just one phosphatidylcholine-associated NSAID has already been approved for commercialization. Nevertheless, the preclinical and clinical data obtained so far indicate that both strategies may improve the safety of nonsteroidal anti-inflammatory therapy.

Evolving therapeutic strategies to improve nonsteroidal anti-inflammatory drug safety.

Nonsteroidal anti-inflammatory drugs (NSAIDs) possess potent anti-inflammatory and analgesic properties through inhibition of cyclooxygenase enzymes (COX-1 and COX-2), which are responsible for synthesis of proinflammatory mediators. NSAIDs are frequently used for treatment of acute and chronic pain conditions. However, their use is associated with serious dose-dependent gastrointestinal (GI), cardiovascular, renal, and hepatic adverse effects, which pose a serious clinical concern for both patients and physicians. During the past 2 decades, approaches to improving the tolerability of NSAIDs were mainly directed toward discovery of COX-2 selective NSAIDs (coxibs), which were expected to minimize the risk of GI injury. Unfortunately, the results from multiple clinical studies have shown that treatment with coxibs may increase the risk for cardiovascular complications. This review summarizes current strategies used to reduce the toxicity of NSAIDs and outlines novel therapeutic approaches still in preclinical development. To minimize the risk of GI ulcerations and bleeding, combination therapies with gastroprotective agents are currently recommended. The new therapeutic agents anticipated to have similar effects include nitric oxide- and hydrogen sulfide-releasing NSAIDs. Novel manufacturing technologies enhance dissolution and absorption of NSAID products, allowing for their administration at low doses, which could lead to improved drug tolerability without diminishing the analgesic and anti-inflammatory efficacy of NSAIDs. This principle is in line with the current recommendation by the US Food and Drug Administration that NSAIDs should be used at the lowest effective dosage. Finally, NSAID formulations targeted directly to the site of inflammation are expected to reduce systemic drug exposure and thus decrease the risk of systemic adverse effects.

Organ and prenatal toxicity of nonsteroidal anti-inflammatory drugs

Abstract Non-selective cyclooxygenase (COX) inhibitors, commonly referred to as nonsteroidal anti-inflammatory drugs (NSAIDs), are among the most taken pharmaceuticals. In adults, they can have a series of side effects, including especially gastroenterotoxicity, hepatotoxicity, nephrotoxicity, chondrotoxicity, and neurotoxicity, and they can induce allergic reactions. Any exacerbation of symptoms depends on the chemical structure of the drug, its dosage and duration of exposure, individual sensitivity, comorbidities and the degree of inhibition of basic COX isoenzymes - the constitutive (COX-2) and induced (COX-1) expressions. However, data on prenatal toxicity are inconsistent. Classic nonselective COX inhibitors do not result in an increase in the risk of developing significant congenital defects; however, if used in the late-pregnancy period, they can have an adverse effect on the foetus, by inducing the premature closure of the ductus arteriosus and by producing a tocolytic effect. Individual reports also indicate the increased risk of developing heart and anterior abdominal wall defects, as well as hypospadias.

Abstract 1903: Application of lipidomics to a sulindac intervention of aromatase inhibitor-induced pain

Abstract (a) Introduction. The regular use of non-steroidal anti-inflammatory drugs (NSAIDs) has been associated with a lower risk for epithelial cancers including breast. NSAIDs block cyclooxygenase (COX)-1 and -2 enzyme metabolism of Ω-6 polyunsaturated fatty acids (PUFA) to prostaglandins (PGs). Additionally, Ω-6 and Ω-3 PUFA are metabolized by lipoxygenases (LOX) and cytochrome P450 (CYP450) enzyme families to produce over 100 metabolites known as ‘oxylipins’. Oxylipins exhibit a wide spectrum of biological activity including mediators of pain and inflammation. The overarching objective of the work is to determine if sulindac (an NSAID) intervention changes oxylipin profiles in breast cancer patients, and if individual oxylipins are related to pain caused by aromatase inhibitors (AI). (b) Methods. This work takes place within the context of an R01-funded single arm, open-label clinical trial of sulindac (N = 100). Breast cancer patients that are stable on AIs complete pain and quality of life questionnaires, and provide blood and urine samples after a 4-week NSAID washout, after a 3 month observation period, at baseline and 3, 6 and 12 months on sulindac intervention (150 mg bid). Oxylipins were quantified in blood and urine (n = 10) at the end of washout, baseline, and after 3 months on sulindac using liquid chromatography (LC) mass spectrometry (MS)-based methods. (c) Results. We quantified 62 total oxylipins in plasma and 87 in urine. Thus far, our analysis has focused on plasma oxylipins. Wilcoxon signed-rank tests were used to compare oxylipin levels at baseline and after 3 months of sulindac. COX metabolites of both Ω-6 and Ω-3 PUFA were non-significantly decreased. In terms of Ω-6 metabolites, 7 LOX products were significantly decreased 11-HETE (P = 0.028), 15-HETE (P = 0.037), 9,12,13-TriHOME (P = 0.017), 9,10,13-TriHOME (P = 0.012), 8-HETE (P = 0.028), 9-HETE (P = 0.028), and 12-HETE (P = 0.047). Significantly decreased Ω-6 CYP450 metabolites were 15(S)-HETrE (P = 0.028), and 5,6-DiHETrE (P = 0.028). The Ω-3 metabolites that significantly decreased were the LOX-derived 5-HEPE (P = 0.022), and 15-HEPE (P = 0.028), and CYP450-derived 5,15-DiHETE (P = 0.047), 5,6-DiHETE (P = 0.009), and 15,16-DiHODE (P = 0.012). Thus far 53% of participants reported pain other than “every day types of pain” post-washout. The Ω-6 CYP450 metabolite 11(12)-EpETrE was significantly correlated with both “worst pain” (ρ = -0.669; P = 0.034) and severity (ρ = -0.778; P = 0.008). The Ω-6 CYP450 metabolite 20-HETE was significantly correlated with severity (ρ = -0.714; P = 0.047). (d) Conclusions. Our preliminary data indicate that oxylipin profiles change in response to sulindac and may be related to pain. Success in shifting plasma oxylipin profiles toward an anti-inflammatory/anti-thromobotic/cardioprotective profile would maximize efficacy and to reduce potential toxicities of NSAIDs within the context of for prevention of breast and other cancers. Citation Format: Jessica Anne Miller, Betsy Werthiem, Jun Yang, Bruce Hammock, Denise Roe, Alison Stopeck, Patricia A. Thompson. Application of lipidomics to a sulindac intervention of aromatase inhibitor-induced pain. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1903. doi:10.1158/1538-7445.AM2015-1903

Non-Helicobacter pylori, Non-NSAID Peptic Ulcer Disease: An Important Consideration in the Evaluation of Patients with Gastric or Duodenal Ulcers

Peptic ulcers are common although the pattern of aetiology is changing. The traditionally most frequent causes of Helicobacter pylori infection and non-steroidal anti-inflammatory toxicity are likely to assume less relative importance as the proportion of cases attributed to other causes rises. Primary care physicians should be aware the pathogenesis of peptic ulceration in non-H. pylori non-NSAID induced disease. This article considers the various less prevalent causes of peptic ulcers and outlines their evaluation and management. Greater awareness of such conditions will improve patient care when traditional causes and remedies are absent.

Hepatotoxicity and Changes in Liver Enzymes Due to Use of Non-Steroidal Anti Inflammatory Drugs (NSAIDs) in Non-Traumatic Musculoskeletal Painful Disorders

The present study aims at critical analysis and investigation of liver function test and hepatotoxity with use of NSAIDs in patients of Non- Traumatic musculoskeletal painful disorders. NSAIDs are most commonly used by medical fraternity worldwide for relief from acute, recurring or chronic pain conditions and are easily available in India in many forms over-the-counter analgesic, sometimes resulting in adverse drug reactions (ADR). The use of NSAIDs and gastrointestinal and renal disturbance is well known. The most frequent ADR due to NSAIDs use was gastrointestinal effects leading to discontinuation of treatment The liver plays a central role in drug metabolism. However, little is known about the cause and effect of NSAIDs on liver and liver enzymes. Most NSAIDs induce hepatic toxicity are due to individual patient susceptibility (idiosyncrasy), and intrinsic associated with mainly hepatocellular tissue. Hepatic toxicity response ranges from asymptomatic or transient derangement in liver function test to fulminant liver damage. The hepatotoxicity due to risk factors associated with NSAID toxicity must be kept in mind by physicians whenever prescribing NSAIDs, because much of underreporting is usually due to transient and asymptomatic effects on liver.

Non-steroidal anti-inflammatory drug use in chronic pain conditions with special emphasis on the elderly and patients with relevant comorbidities: management and mitigation of risks and adverse effects.

Non-steroidal anti-inflammatory drugs (NSAIDs) are among the most frequently used drugs, and this widespread use is complicated by safety issues. A Literature review was conducted. NSAIDs are a leading cause of drug-related morbidity, especially in the elderly and patients with comorbidities. Most adverse effects are related to generalized inhibition of the major targets of NSAIDs: cyclooxygenases I and II. These enzymes are not only involved in pain and inflammation pathogenesis but are also required in the gastrointestinal (GI) tract for mucosal protection and gut motility, and in the kidneys for functional integrity. Thus, the mechanisms of NSAID toxicity are well understood, but the consequences are largely uncontrolled in clinical practice. GI ulcers, including bleeding ulcers, may occur in several percent of all chronic unprotected, high-dose NSAID users. Renal side effects may precipitate renal failure, resulting in acute dialysis and chronic retention. This includes sodium retention, resulting in arterial hypertension, heart failure, and atherosclerotic events. Cardiovascular risk may be tripled by chronic high-dose NSAID use in long-term clinical trials though "real-life studies" indicate lower risk ratios. Off-target side effects include allergic reactions, drug-induced liver injury, and central nervous system effects. Management of pain and inflammation must consider those risks and find alternative drugs or approaches to limit the negative impact of NSAIDs on mortality and morbidity. Alternative drugs, low-dose/short-term use, but especially non-pharmacologic approaches, such as physiotherapy, exercise, neurophysiologic measures, and local therapies, need to be further utilized. The appalling equation "less pain-more deaths/morbidity" ultimately necessitates treatment optimization in the individual patient.

Nonsteroidal anti-inflammatory drugs and cardiovascular outcomes in women: results from the women's health initiative.

Conclusive data about cardiovascular toxicity of nonsteroidal anti-inflammatory drugs (NSAIDs) are sparse. We hypothesized that regular NSAID use is associated with increased risk for cardiovascular events in postmenopausal women, and that this association is stronger with greater cyclooxygenase (cox)-2 when compared with cox-1 inhibition. Postmenopausal women enrolled in the Women's Health Initiative were classified as regular users or nonusers of nonaspirin NSAIDs. Cox regression examined NSAID use as a time-varying covariate and its association with the primary outcome of total cardiovascular disease defined as cardiovascular death, nonfatal myocardial infarction, or nonfatal stroke. Secondary analyses considered the association of selective cox-2 inhibitors (eg, celecoxib), nonselective agents with cox-2>cox-1 inhibition (eg, naproxen), and nonselective agents with cox-1>cox-2 inhibition (eg, ibuprofen) with the primary outcome. Overall, 160 801 participants were available for analysis (mean follow-up, 11.2 years). Regular NSAID use at some point in time was reported by 53 142 participants. Regular NSAID use was associated with an increased hazard for cardiovascular events versus no NSAID use (hazard ratio [HR], 1.10; 95% confidence interval, 1.06-1.15; P<0.001). Selective cox-2 inhibitors were associated with a modest increased hazard for cardiovascular events (hazard ratio, 1.13; 1.04-1.23; P=0.004 and celecoxib only: HR, 1.13; 1.01-1.27; P=0.031). Among aspirin users, concomitant selective cox-2 inhibitor use was no longer associated with increased hazard for cardiovascular events. There was an increased risk for agents with cox-2>cox-1 inhibition (HR, 1.17; 1.10-1.24; P<0.001 and naproxen only: HR, 1.22; 1.12-1.34; P<0.001). This harmful association remained among concomitant aspirin users. We did not observe a risk elevation for agents with cox-1>cox-2 inhibition (HR, 1.01; 0.95-1.07; P=0.884 and ibuprofen only: HR, 1.00; 0.93-1.07; P=0.996). Regular use of selective cox-2 inhibitors and nonselective NSAIDs with cox-2>cox-1 inhibition showed a modestly increased hazard for cardiovascular events. Nonselective agents with cox-1>cox-2 inhibition were not associated with increased cardiovascular risk. www.clinicaltrials.gov. Unique identifier: NCT00000611.

Tu1230 Comparative Proteome Analysis of Indomethacin-Treated Intestinal Cells Reveals Down-Regulation of Collagen I in NSAID-Induced Intestinal Injury

Background: Small intestinal lesions induced by NSAIDs are of great concern in clinical settings. Various hypotheses, mainly of which related to the inhibition of prostaglandin synthesis, have been proposed for the origin of these inflammatory responses, however, the precise mechanism is yet to be known. The cellular process of the lesions must involve upand down-regulations of a large number of proteins and complex interactions between them. To elucidate it, global and systematic identification of the proteins in intestinal cells affected by non-steroidal anti-inflammatory drugs (NSAIDs) is essential. Two-dimensional polyacrylamide gel electrophoresis allows the analysis of total proteins from both cells and tissues, so it can help to elucidate the complex interactions and mechanisms inside cells. Aims: To identify the proteins that play a critical role in NSAIDs toxicity in intestinal cells and to clarify the mechanism of intestinal mucosal injuries induced by NSAIDs. Materials and Methods: We performed 2-DE on IEC-6 rat normal intestinal cells that were treated with indomethacin (200 μΜ, 24 h) or a vehicle control. The differentially displayed proteins were identified through matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Results: We found 18 up-regulated and 8 down-regulated proteins that showed a statistically significant difference of p < 0.05 and a ≥1.5 average fold difference in spot volume as a result of indomethacin treatment. Among these, we found that collagen I and the proteins involved in collagen I synthesis and maturation were all down-regulated. Consistent with this, immunohistochemical analysis showed that the indomethacin-treated rat intestinal mucosal cells exhibits decreased collagen I expression on its apical surface. Furthermore, the cell-protective effect of collagen I on intestinal mucosal cells was demonstrated by the use of a collagen-synthesis inhibitor and cell cultivation on collagen-coated plates versus uncoated plates. Conclusions: For the first time in the literature, a proteomic approach was employed to identify the global proteome alterations of the intestinal mucosa using indomethacin-treated and untreated IEC-6 cells. These results show that collagen I may play an important role in cytoprotection against indomethacin-induced intestinal mucosal injury. Insights gained from the current study may facilitate the development of new therapeutic strategies against NSAID-induced intestinal injury.

Tu1232 Expressions of CYP3A and P-Glycoprotein Were Decreased in the Secondary Inflammatory Response of Dextran Sodium Sulfate-Induced Mice Colitis and Its Contribution to the Blood Concentration of Cyclosporine A

Background: Small intestinal lesions induced by NSAIDs are of great concern in clinical settings. Various hypotheses, mainly of which related to the inhibition of prostaglandin synthesis, have been proposed for the origin of these inflammatory responses, however, the precise mechanism is yet to be known. The cellular process of the lesions must involve upand down-regulations of a large number of proteins and complex interactions between them. To elucidate it, global and systematic identification of the proteins in intestinal cells affected by non-steroidal anti-inflammatory drugs (NSAIDs) is essential. Two-dimensional polyacrylamide gel electrophoresis allows the analysis of total proteins from both cells and tissues, so it can help to elucidate the complex interactions and mechanisms inside cells. Aims: To identify the proteins that play a critical role in NSAIDs toxicity in intestinal cells and to clarify the mechanism of intestinal mucosal injuries induced by NSAIDs. Materials and Methods: We performed 2-DE on IEC-6 rat normal intestinal cells that were treated with indomethacin (200 μΜ, 24 h) or a vehicle control. The differentially displayed proteins were identified through matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Results: We found 18 up-regulated and 8 down-regulated proteins that showed a statistically significant difference of p < 0.05 and a ≥1.5 average fold difference in spot volume as a result of indomethacin treatment. Among these, we found that collagen I and the proteins involved in collagen I synthesis and maturation were all down-regulated. Consistent with this, immunohistochemical analysis showed that the indomethacin-treated rat intestinal mucosal cells exhibits decreased collagen I expression on its apical surface. Furthermore, the cell-protective effect of collagen I on intestinal mucosal cells was demonstrated by the use of a collagen-synthesis inhibitor and cell cultivation on collagen-coated plates versus uncoated plates. Conclusions: For the first time in the literature, a proteomic approach was employed to identify the global proteome alterations of the intestinal mucosa using indomethacin-treated and untreated IEC-6 cells. These results show that collagen I may play an important role in cytoprotection against indomethacin-induced intestinal mucosal injury. Insights gained from the current study may facilitate the development of new therapeutic strategies against NSAID-induced intestinal injury.

Tu1234 Effects of the Dipeptidyl Peptidase-4 Inhibitor, Sitagliptin, on Indomethacin-Induced Intestinal Mucosal Injury in a Model of Type 2 Diabetes

Background: Small intestinal lesions induced by NSAIDs are of great concern in clinical settings. Various hypotheses, mainly of which related to the inhibition of prostaglandin synthesis, have been proposed for the origin of these inflammatory responses, however, the precise mechanism is yet to be known. The cellular process of the lesions must involve upand down-regulations of a large number of proteins and complex interactions between them. To elucidate it, global and systematic identification of the proteins in intestinal cells affected by non-steroidal anti-inflammatory drugs (NSAIDs) is essential. Two-dimensional polyacrylamide gel electrophoresis allows the analysis of total proteins from both cells and tissues, so it can help to elucidate the complex interactions and mechanisms inside cells. Aims: To identify the proteins that play a critical role in NSAIDs toxicity in intestinal cells and to clarify the mechanism of intestinal mucosal injuries induced by NSAIDs. Materials and Methods: We performed 2-DE on IEC-6 rat normal intestinal cells that were treated with indomethacin (200 μΜ, 24 h) or a vehicle control. The differentially displayed proteins were identified through matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Results: We found 18 up-regulated and 8 down-regulated proteins that showed a statistically significant difference of p < 0.05 and a ≥1.5 average fold difference in spot volume as a result of indomethacin treatment. Among these, we found that collagen I and the proteins involved in collagen I synthesis and maturation were all down-regulated. Consistent with this, immunohistochemical analysis showed that the indomethacin-treated rat intestinal mucosal cells exhibits decreased collagen I expression on its apical surface. Furthermore, the cell-protective effect of collagen I on intestinal mucosal cells was demonstrated by the use of a collagen-synthesis inhibitor and cell cultivation on collagen-coated plates versus uncoated plates. Conclusions: For the first time in the literature, a proteomic approach was employed to identify the global proteome alterations of the intestinal mucosa using indomethacin-treated and untreated IEC-6 cells. These results show that collagen I may play an important role in cytoprotection against indomethacin-induced intestinal mucosal injury. Insights gained from the current study may facilitate the development of new therapeutic strategies against NSAID-induced intestinal injury.

Tu1231 Inhibition of RAC-1 Ameliorates Dextran Sodium Sulfate-Induced Colitis in Mice

Background: Small intestinal lesions induced by NSAIDs are of great concern in clinical settings. Various hypotheses, mainly of which related to the inhibition of prostaglandin synthesis, have been proposed for the origin of these inflammatory responses, however, the precise mechanism is yet to be known. The cellular process of the lesions must involve upand down-regulations of a large number of proteins and complex interactions between them. To elucidate it, global and systematic identification of the proteins in intestinal cells affected by non-steroidal anti-inflammatory drugs (NSAIDs) is essential. Two-dimensional polyacrylamide gel electrophoresis allows the analysis of total proteins from both cells and tissues, so it can help to elucidate the complex interactions and mechanisms inside cells. Aims: To identify the proteins that play a critical role in NSAIDs toxicity in intestinal cells and to clarify the mechanism of intestinal mucosal injuries induced by NSAIDs. Materials and Methods: We performed 2-DE on IEC-6 rat normal intestinal cells that were treated with indomethacin (200 μΜ, 24 h) or a vehicle control. The differentially displayed proteins were identified through matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Results: We found 18 up-regulated and 8 down-regulated proteins that showed a statistically significant difference of p < 0.05 and a ≥1.5 average fold difference in spot volume as a result of indomethacin treatment. Among these, we found that collagen I and the proteins involved in collagen I synthesis and maturation were all down-regulated. Consistent with this, immunohistochemical analysis showed that the indomethacin-treated rat intestinal mucosal cells exhibits decreased collagen I expression on its apical surface. Furthermore, the cell-protective effect of collagen I on intestinal mucosal cells was demonstrated by the use of a collagen-synthesis inhibitor and cell cultivation on collagen-coated plates versus uncoated plates. Conclusions: For the first time in the literature, a proteomic approach was employed to identify the global proteome alterations of the intestinal mucosa using indomethacin-treated and untreated IEC-6 cells. These results show that collagen I may play an important role in cytoprotection against indomethacin-induced intestinal mucosal injury. Insights gained from the current study may facilitate the development of new therapeutic strategies against NSAID-induced intestinal injury.

Oxidative stress and liver damage in birds exposed to diclofenac and lead

Responses of wildlife to multiple stressors fit in the ecological concept of trade-off. While toxicity of non-steroidal anti-inflammatory drugs and heavy metals for free-ranging birds has been shown in single exposures, the present study aims to evaluate oxidative stress, and liver and kidney damage caused by single and combined effects of diclofenac and lead in the Japanese quail. Forty Japanese quail (Coturnix coturnix japonica) were divided into equal groups of controls, diclofenac, Pb, and Pb+diclofenac exposures. The birds were exposed to the respective chemicals through insertion of lead shots (1.5 g) into the crop on day 0 of the experiment and/or administration of 5 mg/kg of diclofenac intramuscularly in two treatments on days 0 and 5. Groups in liver and kidney tissues of birds were then compared after 10 days using histopathology and biochemistry markers such as glutathione reductase (GR), ferric reducing antioxidant power (FRAP), and lipid peroxidation measured as total thiobarbituric acid reactive species (TBARS). The liver damage score gradient was Pb+diclofenac exposure group &gt; Pb exposure group &gt; diclofenac exposure group and hepatic TBARS values were significantly increased in the group of birds exposed to a combination of diclofenac and lead compared to the healthy control group. The study has shown that, apart from the reported nephrotoxicity of diclofenac, hepatic toxicity should also be considered. Avian clinicians should be cautious when selecting drugs for therapy of wild birds with unknown history of exposure to toxic substances.

Abstract A15: Chemopreventive efficacy of Phospho-Ibuprofen, Rosuvastatin and their combination on AOM-induced colon adenocarcinoma formation in F344 rats

Abstract Phospho-Ibuprofen (P-I) is a derivative of ibuprofen designed to reduce the traditional toxicities of non-steroidal anti-inflammatory drugs (NSAIDs). P-I is studied for its anti-inflammatory and cancer properties in arthritis, breast cancer models and showed better gastrointestinal safety compared to NSAIDs. Statins has been effective for cholesterol lowering and protecting cardiovascular diseases in addition to its antineoplastic effects in preclinical and clinical observations. A promising strategy to enhance the cancer preventive efficacy of statins and NSAIDs in combination is to produce synergy without any unwanted side effects. Hence, experiments were designed to evaluate the chemopreventive efficacies of P-I and Rosuvastatin, which were administered individually and in combination on azoxymethane (AOM)-induced colon cancer formation in F344 rats. Male F344 rats (~30-45 animals/group) were fed AIN-76A diet and colon tumors were induced with two weekly doses of AOM (15 mg/kg body wt). Eight weeks after the AOM-treatment, groups of rats were fed AIN-76A diets containing ibuprofen (200 ppm), P-I (360 and 720 ppm), Rosuvastatin (50 and 100 ppm), or their low dose combinations for 40 weeks. Colonic tumors were evaluated histopathologically as invasive and noninvasive adenocarcinomas (AdCa) and modulating effects of P-I, and Rosuvastatin and combinations were studied on markers of colon tumor cell proliferation and apoptosis. Results: Administration of P-I at 720 ppm significantly suppressed AOM-induced colon non-invasive and invasive AdCa formation (multiplicity (Mean±SEM): Non-invasive AdCa: Untreated, 1.37±0.22; P-I treated, 0.70±0.14 (p&amp;lt;0.006); Invasive AdCa, Untreated, 2.22±0.35; P-I treated, 1.27±0.20 (p&amp;lt;0.01). Rosuvastatin showed significant inhibition of noninvasive AdCa [50 ppm 1.43±0.21 (p&amp;lt;0.03); 100 ppm 1.06±0.18 (p&amp;lt;0.003)]. Importantly, low dose combination of 360 ppm P-I + 50 ppm Rosuvastatin showed significant synergistic/additive effect on the inhibition of colon non-invasive (44.5%, p&amp;lt;0.016) and invasive AdCa (58.2%, p&amp;lt;0.0002) multiplicity, suggesting a more pronounced effect on invasive AdCa. Whereas administration of 200 ppm Ibuprofen equalant to 360 ppm P-I failed to show any significant protective (p&amp;gt;0.05) effect on AOM-induced AdCa multiplicity or incidence. Proliferation markers, proliferating cell nuclear antigen (PCNA), Cdk2, Cdc25C and Cav1, in tumors of rats exposed to P-I, Rosuvastatin and/or combinations were significantly suppressed. Also, colonic tumors from rats fed combination diet showed significant suppression of Erk and pErk protein expression as compared to colon tumors from rats fed the control diet. This is the first report on the combination using low-dose P-I and Rosuvastatin, which significantly suppressed the colon adenocarcinomas. Our results suggest that low-dose P-I with Rosuvastatin might potentially be a useful combination for colon cancer prevention/treatment in high risk individuals. (Supported by NCI-CN-N01-53300). Citation Format: Naveena B. Janakiram, Altaf Mohammed, Yuting Zhang, Misty Brewer, Taylor Bryant, Laura Biddick, Stan Lightfoot, Vernon E. Steele, Chinthalapally V. Rao. Chemopreventive efficacy of Phospho-Ibuprofen, Rosuvastatin and their combination on AOM-induced colon adenocarcinoma formation in F344 rats. [abstract]. In: Proceedings of the Twelfth Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2013 Oct 27-30; National Harbor, MD. Philadelphia (PA): AACR; Can Prev Res 2013;6(11 Suppl): Abstract nr A15.

Non-Steroidal Anti-Inflammatory Drugs (NSAIDs) are Associated with a High Incidence of Gastrointestinal Side-Effects

Inhibition of the cyclooxygenase &nbsp;(COX) enzyme is the basis for both the efficacy and toxicity of NSAIDs. The aim of this study was to avaluate the non-steroidal anti-inflammatory drugs were used in neuro polyclinic hospital of Dr. Mintohardjo Jakarta, and to evaluate gastrointestinal tolerability of meloxicam 15 mg compared with diclofenac sodium 100 mg. The methode of this study was cross-sectional observation and&nbsp; cohort prospective on December 2010-March 2011. The data of dyspepsia associated were used non-steroidal anti-inflammatory drugs consist of pain in upper abdomen, nausea, vomiting, upper &nbsp;abdominal bloating and early satiety collected with PADYQ (The porto alegre dyspeptic symptoms questionnaire) were assessed at baseline and after 2 and 4 weeks of treatment. The non-steroidal anti-inflammatory drugs used in neuro polyclinic hospital of Dr. Mintohardjo Jakarta were meloxicam (48.21%), diclofenac sodium (31.07%), mefenamic acid (15.36%), piroxicam (3.93%)&nbsp; dan acetaminophen (1.43%). Insiden of adverse event after 2 weeks treatment was significantly lower in the meloxicam group compared with diclofenac sodium group in pain in upper abdomen and&nbsp;upper abdominal bloating (p=0.020 and&nbsp; p=0.037). These result suggest that meloxicam was much better tolerated than diclofenac sodium after 2 weeks treatment.