Background: Earlier studies reported the anti-inflammatory activity in several species of Piper, and Piper umbellatum Linn. leaves containing some phytochemicals that are potent anti-inflammatory agents. However, there was no thorough investigation on the anti-inflammatory activity of the locally grown P. umbellatum in the Philippines. Objective: The study aimed to determine the anti-inflammatory activity of Piper umbellatum leaves using in vitro and in vivo assays. Methodology: Crude extracts were obtained from P. umbellatum leaves using polar and non-polar solvents. The anti-inflammatory activities of all crude extracts were determined using the carrageenan-induced paw edema test in mice and phytochemical analysis. The crude extract with the highest activity was partially purified using column chromatography. The fractions with similar TLC profile were pooled and tested for antiinflammatory activity. COX-1 and COX-2 enzyme inhibitory activity were determined in pooled fractions that showed initial activity in animal model. Results: Among the crude extracts of P.umbellatum, the crude ethyl acetate extract exhibited a significant dose-dependent inhibition on paw edema test with doses of 500 mg/kg bw, 1,000 mg/kg bw and 1,500 mg/kg bw (p<0.05). Among the 20 pooled fractions (PF) collected from the ethyl acetate extract, PF58, PF60 and PF64 had the highest COX-2 enzyme inhibitions of 83.12 %, 84.78% and 77.47%, respectively (p<0.05). PF60 also exhibited the highest anti-inflammatory activity on paw edema with inhibitions of 62.45% at low dose (250 mg/kg bw) and 76.10 % at high dose (1,000 mg/kg bw) in mice. Conclusion The ethyl acetate extract of P. umbellatum leaves and its fraction-PF60 exhibited a significant anti-inflammatory activity in in vitro and in vivo assays and contained high amounts of total phenolic and total flavonoid.
Prostaglandin-Endoperoxide Synthases ; Carrageenan ; Inflammation

BACKGROUND/OBJECTIVES: Anti-inflammatory and antioxidative activities of luteolin and luteolin-7-O-glucoside were compared in galactosamine (GalN)/lipopolysaccharide (LPS)-induced hepatitic ICR mice. MATERIALS/METHODS: Male ICR mice (6 weeks old) were divided into 4 groups: normal control, GalN/LPS, luteolin, and luteolin-7-O-glucoside groups. The latter two groups were administered luteolin or luteolin-7-O-glucoside (50 mg/kg BW) daily by gavage for 3 weeks after which hepatitis was induced by intraperitoneal injection of GalN and LPS (1 g/kg BW and 10 µg/kg BW, respectively). RESULTS: GalN/LPS produced acute hepatic injury by a sharp increase in serum AST, ALT, and TNF-α levels, increases that were ameliorated in the experimental groups. In addition, markedly increased expressions of cyclooxygenase (COX)-2 and its transcription factors, nuclear factor (NF)-κB and activator protein (AP)-1, were also significantly attenuated in the experimental groups. Compared to luteolin-7-O-glucoside, luteolin more potently ameliorated the levels of inflammatory mediators. Phase II enzymes levels and NF-E2 p45-related factor (Nrf)-2 activation that were decreased by GalN/LPS were increased by luteolin and luteolin-7-O-glucoside administration. In addition, compared to luteolin, luteolin-7-O-glucoside acted as a more potent inducer of changes in phase II enzymes. Liver histopathology results were consistent with the mediator and enzyme results. CONCLUSION: Luteolin and luteolin-7-O-glucoside protect against GalN/LPS-induced hepatotoxicity through the regulation of inflammatory mediators and phase II enzymes.
Animals ; Galactosamine ; Hepatitis ; Humans ; Inflammation ; Injections, Intraperitoneal ; Liver ; Luteolin ; Male ; Mice ; Mice, Inbred ICR ; NF-E2-Related Factor 2 ; NF-kappa B ; Prostaglandin-Endoperoxide Synthases ; Transcription Factors

BACKGROUND: Brennan’s rodent paw incision model has been extensively used for understanding mechanisms underlying postoperative pain in humans. However, alterations of physiological parameters like blood pressure and heart rate, or even feeding and drinking patterns after the incision have not been documented as yet. Moreover, though eicosanoids like prostaglandins and leukotrienes contribute to inflammation, tissue levels of these inflammatory mediators have never been studied. This work further investigates the antinociceptive effect of protein C after intra-wound administration. METHODS: Separate groups of Sprague–Dawley rats were used for quantitation of cyclooxygenase (COX) activity and leukotriene B4 level by enzyme-linked immunosorbent assay, as well as estimation of cardiovascular parameters and feeding and drinking behavior after paw incision. In the next part, rats were subjected to incision and 10 μg of protein C was locally administered by a micropipette. Both evoked and non-evoked pain parameters were then estimated. RESULTS: COX, particularly COX-2 activity and leukotriene B4 levels increased after incision. Hemodynamic parameters were normal. Feeding and drinking were affected on days 1 and 3, and on day 1, respectively. Protein C attenuated non-evoked pain behavior alone up to day 2. CONCLUSIONS: Based upon current observations, Brennan’s rodent paw incision model appears to exhibit a prolonged period of nociception similar to that after surgery, with minimal interference of physiological parameters. Protein C, which is likely converted to activated protein C in the wound, attenuated the guarding score, which probably represents pain at rest after surgery in humans.
Animals ; Blood Pressure ; Drinking ; Drinking Behavior ; Eicosanoids ; Enzyme-Linked Immunosorbent Assay ; Heart Rate ; Hemodynamics ; Humans ; Inflammation ; Leukotriene B4 ; Leukotrienes ; Nociception ; Pain, Postoperative ; Prostaglandin-Endoperoxide Synthases ; Prostaglandins ; Protein C ; Rats ; Rodentia ; Wounds and Injuries

Ginger, one of worldwide consumed dietary spice, is not only famous as food supplements, but also believed to exert a variety of remarkable pharmacological activity as herbal remedies. In this study, a ginger constituent, 12-dehydrogingerdione (DHGD) was proven that has comparable anti-inflammatory activity with positive control 6-shogaol in inhibiting LPS-induced interleukin (IL)-6, tumor necrosis factor (TNF)-α, prostaglandin (PG) E₂, nitric oxide (NO), inducible NO synthase (iNOS) and cyclooxygenase (COX)-2, without interfering with COX-1 in cultured microglial cells. Subsequent mechanistic studies indicate that 12-DHGD may inhibit neuro-inflammation through suppressing the LPS-activated Akt/IKK/NF-κB pathway. Furthermore, 12-DHGD markedly promoted the activation of NF-E2-related factor (Nrf)-2 and heme oxygenase (HO)-1, and we demonstrated that the involvement of HO-1 on the production of pro-inflammatory mediators such as NO and TNF-α by using a HO-1 inhibitor, Zinc protoporphyrin (Znpp). These results indicate that 12-DHGD may protect against neuro-inflammation by inhibiting Akt/IKK/IκB/NF-κB pathway and promoting Nrf-2/HO-1 pathway.
Dietary Supplements ; Ginger ; Heme Oxygenase (Decyclizing) ; Interleukins ; Microglia* ; Nitric Oxide ; Nitric Oxide Synthase ; Prostaglandin-Endoperoxide Synthases ; Spices ; Tumor Necrosis Factor-alpha ; Zinc

Neuroinflammation is one of the key mechanisms of neuropathic pain, which is primarily mediated by the Toll-like receptor 4 (TLR4) signaling pathways in microglia. Therefore, TLR4 may be a reasonable target for treatment of neuropathic pain. Here, we examined the analgesic effect of TLR4 antagonistic peptide 2 (TAP2) on neuropathic pain induced by spinal nerve ligation in rats. When lipopolysaccharide (LPS)-stimulated BV2 microglia cells were treated with TAP2 (10 µM), the mRNA levels of proinflammatory mediators, such as tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, cyclooxygenase (COX)-2, and inducible nitric oxide synthase (iNOS), were markedly decreased by 54–83% as determined by quantitative PCR (qPCR) analysis. Furthermore, when TAP2 (25 nmol in 20 µL PBS) was intrathecally administered to the spinal nerve ligation-induced rats on day 3 after surgery, the mechanical allodynia was markedly decreased for approximately 2 weeks in von Frey filament tests, with a reduction in microglial activation. On immunohistochemical and qPCR analyses, both the level of reactive oxygen species and the gene expression of the proinflammatory mediators, such as TNF-α, IL-1β, IL-6, COX-2, and iNOS, were significantly decreased in the ipsilateral spinal dorsal horn. Finally, the analgesic effect of TAP2 was reproduced in rats with monoiodoacetate-induced osteoarthritic pain. The findings of the present study suggest that TAP2 efficiently mitigates neuropathic pain behavior by suppressing microglial activation, followed by downregulation of neuropathic pain-related factors, such as reactive oxygen species and proinflammatory molecules. Therefore, it may be useful as a new analgesic for treatment of neuropathic pain.
Analgesics ; Animals ; Down-Regulation ; Gene Expression ; Hyperalgesia ; Interleukin-6 ; Interleukins ; Ligation ; Microglia ; Neuralgia ; Nitric Oxide Synthase Type II ; Polymerase Chain Reaction ; Prostaglandin-Endoperoxide Synthases ; Rats ; Reactive Oxygen Species ; RNA, Messenger ; Spinal Cord Dorsal Horn ; Spinal Nerves ; Toll-Like Receptor 4 ; Toll-Like Receptors ; Tumor Necrosis Factor-alpha

The purpose of this study was to evaluate the effect of mangosteen extract complex (MEC; Garcinia mangostana L. and propolis extracts) on the inhibition of inflammation and prevention of alveolar bone loss using a ligature-induced periodontitis model. Rat molars were ligatured with silk, and 1 µg/mL of lipopolysaccharide of Porphyromonas gingivalis was injected into the buccal and palatal gingivae of the teeth with or without treatment with the MEC. Changes in the expression levels of prostaglandin E₂ (PGE₂), interleukin-8 (IL-8), inducible nitric oxide synthase (iNOS), matrix metalloproteinase-8 (MMP-8), cyclooxygenase (COX)-1, and COX-2 in gingival tissues were evaluated using enzyme-linked immunosorbent assays. Alveolar bone loss around the ligated molars was examined using micro-computed tomography. The expression levels of PGE₂, IL-8, iNOS, MMP-8, COX-1, and COX-2 in gingival tissues were significantly reduced in the group treated with a mixture of 16 µg of mangosteen extract powder and 544 µg of propolis extract powder (ligation [Lig] + lipopolysaccharide extracted from P. gingivalis KCOM 2804 [L] + MEC 1:34). Additionally, alveolar bone loss was significantly reduced in the Lig + L + MEC 1:34 group compared with that in other groups. These results indicate that the MEC could be useful in preventing and treating periodontitis.
Alveolar Bone Loss ; Animals ; Enzyme-Linked Immunosorbent Assay ; Garcinia mangostana ; Garcinia ; Gingiva ; Inflammation ; Interleukin-8 ; Matrix Metalloproteinase 8 ; Molar ; Nitric Oxide Synthase Type II ; Periodontitis ; Porphyromonas gingivalis ; Propolis ; Prostaglandin-Endoperoxide Synthases ; Rats ; Silk ; Tooth

BACKGROUND: Sometimes general anesthesia is required for dental surgery in pregnant women. Facial bone fractures or neck abscess should be treated immediately. Dental surgery, however, creates a stressful situation that can cause inflammation. Inflammatory responses are a well-known major cause of preterm labor and preterm birth. Here we demonstrate the effects of remifentanil on the factors related to preterm labor and its mechanism of action on amniotic-derived epithelial cells (WISH cells). METHODS: WISH cells were exposed to lipopolysaccharide (LPS) for 24 h and co-treated with various concentrations of remifentanil. MTT assays were performed to measure cell viability. To explain the effects of remifentanil on the factors related to inflammation in WISH cells, activation of nuclear factor kappa B (NF-κB) and p38 and the expression of interleukin (IL)-1β, tumor necrosis factor (TNF)-α, cyclooxygenase (COX)2, and prostaglandin E (PGE)2 were quantified using western blotting and RT-PCR, respectively. RESULTS: Remifentanil did not affect WISH cell viability. In western blot analysis, co-treatment with remifentanil resulted in decreased phosphorylation of NF-κB, and expression of COX2 and PGE2 in LPS-induced inflammation, but the results were statistically significant only at low concentrations. Reduction of IL-1β and TNF-α expression was also observed with RT-PCR. CONCLUSION: Co-treatment with remifentanil does not affect the viability of WISH cells, but reduces the expression of the factors related to inflammation, which can induce uterine contraction and preterm labor. These findings provide evidence that remifentanil may inhibit uterine contraction and preterm labor in clinical settings.
Abscess ; Amnion ; Anesthesia, General ; Blotting, Western ; Cell Survival ; Dinoprostone ; Epithelial Cells ; Facial Bones ; Female ; Humans ; Inflammation ; Interleukins ; Neck ; NF-kappa B ; Obstetric Labor, Premature ; Phosphorylation ; Pregnancy ; Pregnant Women ; Premature Birth ; Prostaglandin-Endoperoxide Synthases ; Tumor Necrosis Factor-alpha ; Uterine Contraction

Nonsteroidal anti-inflammatory drugs (NSAIDs) have been the first choice of treatment for axial spondyloarthritis (axSpA); cyclooxygenase (COX)-2 inhibitors reduce both inflammation and bone formation. When NSAID treatment fails, tumor necrosis factor inhibitors (TNFis) can be used to treat active axSpA, but, presently, TNFis cannot completely prevent radiographic progression. Theoretically, as TNF is a strong pro-inflammatory cytokine triggering bone resorption, TNFis should stimulate bone formation. Recently, it was discovered that the IL-23/-17 axis is associated with enthesitis development and bone formation in a mouse model. The anti-IL-23 monoclonal antibody ustekinumab has been approved as treatment for moderate-to-severe plaque psoriasis and psoriatic arthritis. However, in axSpA patients, ustekinumab effectiveness was low and a phase 3 clinical trial was terminated. The anti-IL-17A monoclonal antibody secukinumab has been approved as treatment for moderate-to-severe plaque psoriasis, psoriatic arthritis, and ankylosing spondylitis. In a 3-year extension of the MEASURE 1 observational study, the mean change in modified Stoke Ankylosing Spondylitis Spine Score (mSASSS) from baseline to week 104 was 0.30 ± 2.53 among patients with evaluable X-rays originally randomized to secukinumab (n = 168). Such patients exhibited less radiographic progression compared to other TNFi studies. Several new drugs are in clinical trials exploring their effects on axSpA; these include ixekizumab (an anti-IL-17A monoclonal antibody), brodalumab (an anti-IL-17 receptor monoclonal antibody), and the Janus kinase (JAK) inhibitors tofacitinib and upadacitinib. The IL-23/-17 axis is important in terms of axSpA inflammation and bone formation. However, to date, no drug has completely prevented radiographic progression in axSpA patients.
Animals ; Arthritis, Psoriatic ; Bone Resorption ; Humans ; Inflammation ; Mice ; Observational Study ; Osteogenesis ; Phosphotransferases ; Prostaglandin-Endoperoxide Synthases ; Psoriasis ; Spine ; Spondylitis, Ankylosing ; Tumor Necrosis Factor-alpha ; Ustekinumab

Non-steroidal anti-inflammatory drugs (NSAIDs) are commonly used in various clinical situations, with excellent analgesic, anti-pyretic and anti-inflammatory effects. In addition to gastrointestinal bleeding, which was the first adverse effect to be reported, myriad adverse effects from the digestive system, cardiovascular system, renal system and hematology have been also reported. In early 2000s, a few new cyclooxygenase (COX)-2 selective inhibitors were developed with the expectation of better gastrointestinal safety profile, most of them were withdrawn from the market due to various adverse effects, and interest in safety of NSAIDs has been increased again. Over the past two decades, research on the safety and adverse effects of NSAIDs has accumulated. In brief, celecoxib is associated with fewer gastrointestinal adverse events compared to non-selective NSAIDs. In patients receiving aspirin, the use of non-selective NSAIDs should be avoided, and if an anti-inflammatory drug is required, a COX-2 selective inhibitor should be considered. Celecoxib has been shown to have similar or better safety profile than other non-selective COX inhibitors. Additionally, the new COX-2 selective inhibitors of etorixocib and polmacoxib have been approved. Many factors should be considered when prescribing NSAIDs, as the safety profile of indivisual NSAIDs vary, and NSAIDs have a high risk of duplicate prescription because of the variety of indications and over-the-counter products. Physicians should comprehend the updated guidelines and the results of new clinical studies, and the risk factors for each individual patient should also be reviewed. Physicians should therefore contemplate new prescription strategies.
Anti-Inflammatory Agents, Non-Steroidal ; Aspirin ; Cardiovascular System ; Celecoxib ; Digestive System ; Drug-Related Side Effects and Adverse Reactions ; Hematology ; Hemorrhage ; Humans ; Medication Therapy Management ; Prescriptions ; Prostaglandin-Endoperoxide Synthases ; Risk Factors

BACKGROUND AND OBJECTIVES: Previous studies have shown that integrins alpha5beta1 (ITGA5B1) gene-modified rat bone marrow mesenchymal stem cells (rBMSCs) could prevent cell anoikis and increase the nitric oxide (NO) production. Here we examined the capability of rBMSCs/ITGA5B1 on the phenotype modulation of Human Pulmonary Artery Smooth Muscle Cell (HPASMC) in vitro. METHODS AND RESULTS: The synthetic (dedifferentiated) phenotype of HPASMC was induced by monocrotaline (MCT, 1μM) for 24 h and then co-cultured with rBMSCs/ITGA5B1 in a transwell culture system. The activation of NO/cGMP (nitric oxide/Guanosine-3′, 5′-cyclic monophosphate) signaling was investigated in HPASMC. The changes of pro-inflammatory factors, oxidative stress, vasodilator, vasoconstrictor, contractile and synthetic genes, and the morphological changes of HPASMC were investigated. The results of this study showed that the NO/cGMP signal, endothelial nitric oxide synthase (eNOS) expression, the expression of the vasoprotective genes heme oxygenase-1 (HMOX1) and prostaglandin-endoperoxide synthase 2 (PTGS2) were increased, but the expression of transforming growth factor-β1 (TGF-β1), CCAAT/enhancer-binding proteins delta (Cebpd), Krüppel-like factor 4 (KLF4), and activating transcription factor 4 (ATF4) were reduced in MCT treated HPASMC co-cultured with rBMSCs/ITGA5B1. The synthetic smooth muscle cells (SMCs) phenotype markers thrombospondin-1, epiregulin and the vasoconstrictor endothelin (ET)-1, thromboxane A2 receptor (TbxA2R) were down-regulated, whereas the contractile SMCs phenotype marker transgelin expression was up-regulated by rBMSCs/ITGA5B1. Furthermore, rBMSCs/ITGA5B1 promoted the morphological restoration from synthetic (dedifferentiation) to contractile (differentiation) phenotype in MCT treated HPASMC. CONCLUSIONS: rBMSCs/ITGA5B1 could inhibit inflammation and oxidative stress related genes to promote the HPASMC cell differentiation by activation NO/cGMP signal.
Activating Transcription Factor 4 ; Animals ; Anoikis ; Bone Marrow ; Cell Differentiation ; Endothelins ; Epiregulin ; Genes, Synthetic ; Heme Oxygenase-1 ; Humans ; In Vitro Techniques ; Inflammation ; Integrins ; Mesenchymal Stromal Cells ; Monocrotaline ; Muscle, Smooth, Vascular ; Myocytes, Smooth Muscle ; Nitric Oxide Synthase Type III ; Nitric Oxide ; Oxidative Stress ; Phenotype ; Prostaglandin-Endoperoxide Synthases ; Pulmonary Artery ; Rats ; Receptors, Thromboxane A2, Prostaglandin H2