The Potency of Indonesia’s Pomegranate Peel Ethanol Extract (Punica Granatum Linn.) as Anti-Inflammatory Agent in Mice Colon Induced by Dextran Sodium Sulfate: Focus on Cyclooxygenase-2 and Inos Expressions

Abstract

Objective: Inflammatory disease occurs in the mucosal of the colon, or ulcerative colitis (UC) is one of subtypes of inflammatory bowel disease. The numerous of drug side effects for treatment of colitis give rise to use medicinal herbs as alternative therapies. Pomegranate peel extract has been used for the treatment of pain and inflammatory conditions. This study aimed to investigate the anti-inflammatory effects of pomegranate peel ethanol extract on mice colon through inflammation pathway which reduce inflammation score in mice model of chronic inflammation induced by dextran sodium sulfate (DSS).Methods: Thirty Swiss Webster mice divided randomly into 6 groups: Normal, aspirin 43 mg/kg/d (ASP), ellagic acid 26 mg/kg/d (ELG), DSS 2%b/v (DSS), pomegranate peel ethanol extract 240 mg/kg/d (DOSES-1), and 480 mg/kg/d (DOSES-2). All groups were given DSS 2% over 3 cycles except normal group (where each cycle in the DSS group consisted of 2% DSS in drinking water for 7 days, followed by a 7-day interval with normal drinking water). At the end of the experiment, colon samples were washed with water then buffered neutral formalin 10% fixed and paraffin embedded for histological analysis.Results: DOSES-1 and DOSES-2 were significantly reduced inflammation score in colon mice induced by DSS (p<0.05), mean score 2.01 and 2.02. Expression of cyclooxygenase (COX-2) was significantly decreased (p<0.05), mean score 27.48 and 17.77. Expression of iNOS was also significantly decreased (p<0.05), mean score 54.01 and 36.69. There were no significant differences between DOSES-1 and DOSES-2 groups with ASP and ELG group (p>0.05). Conclusion:Pomegranate peel ethanol extract has an anti-inflammatory agent by reduces inflammation score, inhibiting COX-2 and iNOS expression on mice colon by DSS induced. Furthermore, pomegranate peel ethanol extract has equivalent effectiveness with aspirin and pure ellagic acid.

Anti-Inflammatory Activity of Tinocrisposide by Inhibiting Nitric Oxide Production in Lipopolysaccharides-Stimulated Raw 264.7 Cells

“…Thus, in response to inflammatory stimuli, high level of NO is generated [28]. Exposure of RAW 264.7 cells to LPS upregulates iNOS and pre-treatment of the cell with varying compound concentration (1,5,25,50, and 100 µM) before LPS induction was able to reduce the NO production.…”

“…RAW 264.7 cells were plated in 96-well plate with a density of 10 5 cell/well until confluence. Supernatants were discarded and 100 µl medium containing various concentrations of tinocrisposide (1,5,25,50, and 100 µM) was added to the cells. The cells were allowed to rest for 1 h and then incubated with LPS (1 g/ml) for 15 h at 37°C in a humidified atmosphere containing 5% CO 2.…”

“…This was based on trend seen in NO generated. Meanwhile, the effect to dose used in this study did not affect cell viability since the test dose chosen (1,5,25,50, and 100 µM) was based on cell viability study conducted before NO determination study. Therefore, the NO reduction effect observed is based on tinocrisposide action and not due to a reduction in the cell population.…”

“…Kusmadi et al investigated the anti-inflammatory effects of pomegranate peel ethanol extract on mice colon. The study was done through inflammation pathway and it showed reduction in inflammation score on mice model with chronic inflammation induced by dextran sodium sulfate [25]; meanwhile, Kurian et al explored an anti-inflammatory property of melanin in vitro in RAW 264.7 cell line using cyclooxygenase, lipoxygenase, myeloperoxidase, and cellular nitrite inhibitory assays [26]. The aim of our study was to investigate in vitro the anti-inflammatory activity of tinocrisposide using the LPSstimulated RAW 264.7 macrophage cells.…”

Analysis Reactivity of Punica Granatum Polyphenols to the Osteocalcin, Bone Morphogenetic Protein-2, and Collagen Type-1

“…This active component acts as an antioxidant, anticancer, and also anti-inflammatory by inhibiting proinflammatory cytokines [2]. Moreover, these active components can also inhibit a number of enzymes that play a role in cell differentiation such as cyclooxygenase, lipooxygenase, cytochrome P450, phospholipase A2, ornithine decarboxylase, carbonic anhydrase, 17 beta-hydroxysteroid dehydrogenase, and serine protease [3].…”