ABSTRACT
Asiatic acid, a triterpenoid component isolated from Centella asiatica (L.) Urban, possesses antioxidative and anti-inflammatory activities. In this study, we aimed to investigate the anti-inflammatory effects of asiatic acid both in vivo and in vitro. HGFs or RAW264.7 cells were treated with asiatic acid 1h before LPS treatment. Cell viability was measured by MTT assay. The levels of PGE2, NO, IL-6, and IL-8 were detected by ELISA. Protein expression levels were detected by western blot analysis. In vivo, asiatic acid significantly inhibited LPS-induced IL-6 and IL-8 expression levels in gingival tissues. In vitro, LPS-induced PGE2, NO, IL-6, and IL-8 production was significantly attenuated by asiatic acid. Asiatic acid also inhibited p65 NF-κB phosphorylation induced by LPS in HGFs. The expression of PPAR-γ was up-regulated by asiatic acid. Furthermore, GW9662, a PPAR-γ inhibitor, attenuated the inhibitory effect of asiatic acid on PGE2, NO, IL-6, and IL-8 production. Our results suggest that asiatic acid activates PPAR-γ, which subsequently inhibits LPS-induced NF-κB activation and inflammatory mediators production. Asiatic acid may offer therapeutic potential for the treatment of periodontitis.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Fibroblasts/immunology , Gingiva/pathology , Gingivitis/drug therapy , Macrophages/immunology , Pentacyclic Triterpenes/pharmacology , Anilides/pharmacology , Animals , Centella/immunology , Dinoprostone/metabolism , Fibroblasts/drug effects , Fibroblasts/pathology , Humans , Lipopolysaccharides/immunology , Macrophages/drug effects , Male , Mice , NF-kappa B/metabolism , Nitric Oxide/metabolism , PPAR gamma/antagonists & inhibitors , PPAR gamma/metabolism , RAW 264.7 Cells , Rats , Rats, Sprague-DawleyABSTRACT
Psoriasis is a chronic auto-immune inflammation disease with skin lesions and abnormal keratinocyte proliferation. The IL-23/IL-17 axis plays an important role in the pathogenesis of psoriasis. Madecassoside (MAD) was the most important constituents isolated from Centella asiatica, which has long been used in dermatology, and it is supposed that MAD may have effects on psoriasis. In the present study, the BALB/c mice ear and back skin received IMQ for 6 consecutive days to induce psoriasis-like dermatitis. MAD ointment was applied 6h later after IMQ treatment, and the IL-23/IL-17 pathway was investigated. The HE staining, BrdU and Psoriasis Area and Severity Index (PASI) were used to score the severity of keratinocyte proliferation and inflammation of the skin. Real-time PCR and Western Blot were used to detect the IL-23/IL-17 related cytokines. Flow Cytometry were applied to observe the numbers of Th17 cells. Daily application of IMQ for 6days on mouse ear skin and back skin induced psoriasis-like dermatitis. Real-time PCR showed that mRNA level of IL-23, IL-22, IL-17A were significantly decreased by MAD ointment treatment in ear skin. HE staining and BrdU incorporation implied that MAD ointment reduced keratinocyte proliferation. Flow Cytometry results showed MAD ointment decreased the numbers of Th17 cells. Thus, MAD ointment ameliorates Imiquimod-induced skin inflammation and abnormal keratinocyte through regulate the IL-23/IL-17 axis.
Subject(s)
Centella/immunology , Dermatitis/drug therapy , Interleukin-17/metabolism , Interleukin-23/metabolism , Psoriasis/drug therapy , Skin/drug effects , Th17 Cells/drug effects , Triterpenes/therapeutic use , Aminoquinolines , Animals , Cell Proliferation/drug effects , Disease Models, Animal , Female , Humans , Imiquimod , Mice , Mice, Inbred BALB C , Ointments , Signal Transduction/drug effects , Skin/immunology , Skin/pathology , Th17 Cells/immunologyABSTRACT
Asiatic acid (AA) is one of the major components of Titrated extract of Centella asiatica (TECA), which has been reported to possess antioxidant and anti-inflammatory activities. The purpose of this study was to investigate the protective effect of AA on pulmonary inflammation induced by cigarette smoke (CS). AA significantly attenuated the infiltration of inflammatory cells in bronchoalveolar lavage fluid (BALF) of CS exposure mice. AA also decreased ROS production and NE activity, and inhibited the release of proinflammatory cytokines in BALF. AA reduced the recruitment of inflammatory cells and MCP-1 expression in lung tissue of CS exposure mice. AA also attenuated mucus overproduction, and decreased the activation of MAPKs and NF-kB in lung tissue. Furthermore, AA increased HO-1 expression and inhibited the reduced expression of SOD3 in lung tissue. These findings indicate that AA effectively inhibits pulmonary inflammatory response, which is an important process in the development of chronic obstructive pulmonary disease (COPD) via suppression of inflammatory mediators and induction of HO-1. Therefore, we suggest that AA has the potential to treat inflammatory disease such as COPD.
Subject(s)
Centella/immunology , Drugs, Chinese Herbal/therapeutic use , Lung/drug effects , Neutrophils/drug effects , Pentacyclic Triterpenes/therapeutic use , Pneumonia/drug therapy , Pulmonary Disease, Chronic Obstructive/drug therapy , Animals , Cell Movement/drug effects , Cytokines/metabolism , Heme Oxygenase-1/metabolism , Humans , Inflammation Mediators/metabolism , Lung/metabolism , Lung/pathology , Male , Membrane Proteins/metabolism , Mice , Mice, Inbred C57BL , Mucus/metabolism , Neutrophils/immunology , Pneumonia/chemically induced , Reactive Oxygen Species/metabolism , Smoking/adverse effects , Superoxide Dismutase/metabolismABSTRACT
Recent studies showed that Madecassoside (MAD), a pentacyclic triterpene isolated from Centella asitica (L.), was used as a therapeutic agent in wound healing and also as an anti-inflammatory, anti-oxidative activities and anti-aging agent. However, its role in cancer has not been elucidated. In our present study, hepatocyte growth factor (HGF) induced the phosphorylation of its corresponding receptor cMET, increased expression of cyclo-oxygenase-2 (COX-2) and prostaglandin E2 (PGE2) in human hepatocellular carcinoma (HCC) cells lines (HepG2 and SMMC-77), and this effect was inhibited by MAD in a dose-dependent manner. In addition, MAD exhibited significant anti-proliferative and anti-invasive effect in HGF-induced HepG2 and SMMC-77 cells. Moreover, MAD inhibited the phosphorylation of extracellular signal-regulated kinase 1 and 2 (ERK1/2) and the protein kinase C (PKC) activity in HGF-induced HepG2 and SMMC-77 cells. This conclusion was consistent with the effect of selective COX-2 inhibitor (NS-398) and knockdown of COX-2 by siRNA on attenuating the proliferation and invasiveness potential, and over-expression of COX-2 on abolishing the effects of MAD on proliferation and invasiveness potential, and was also in parallel with the effect of PKC inhibitor (Bisindolylmaleimide) on inhibiting PKC activity, MEK/ERK1/2 inhibitor (PD98059) inhibited MEK/ERK1/2 pathways in HGF-induced HepG2 and SMMC-77 cells. Collectively, MAD could inhibit the HGF-activated proliferation and invasiveness of HCC cells via regulating the activation of cMET-PKC-ERK1/2-COX-2-PGE2 cascade, which indicated that MAD might help control HGF-linked HCC.
Subject(s)
Antineoplastic Agents/pharmacology , Carcinoma, Hepatocellular/drug therapy , Centella/immunology , Liver Neoplasms/drug therapy , Triterpenes/pharmacology , Cell Proliferation/drug effects , Cyclooxygenase 2/genetics , Cyclooxygenase 2/metabolism , Dinoprostone/metabolism , Extracellular Signal-Regulated MAP Kinases/metabolism , Hep G2 Cells , Hepatocyte Growth Factor/metabolism , Humans , Neoplasm Invasiveness , Nitrobenzenes/pharmacology , Protein Kinase C/metabolism , Proto-Oncogene Proteins c-met/metabolism , RNA, Small Interfering/genetics , Signal Transduction/drug effects , Sulfonamides/pharmacologyABSTRACT
In the present study, the effect of asiatic acid, a natural triterpenoid compound, on murine experimental colitis induced by dextran sulfate sodium (DSS) and its possible mechanism were examined in vivo and vitro. Oral administration of asiatic acid dose-dependently attenuated the loss of body weight and shortening of colon length induced by DSS. The disease activity index, histopathologic scores of musco and myeloperoxidase activity were also significantly reduced by asiatic acid treatment. Protein and mRNA levels of DSS-induced pro-inflammatory cytokines in colon, including TNF-α, IL-1ß, IL-6 and IFN-γ, were markedly suppressed by asiatic acid. At the same time, decreased activation of caspase-1 in peritoneal macrophages was detected in asiatic acid-treated mice, which suggested that the NLRP3 inflammasome activation was suppressed. In addition, we also found that asiatic acid dose-dependently inhibited IL-1ß secretion, caspase-1 activation as well as inflammasome assembling in vitro. Furthermore, the mechanism of asiatic acid was related to the inhibition of mitochondrial reactive oxygen species generation and prevention of mitochondrial membrane potential collapse. Taken together, our results demonstrate the ability of asiatic acid to inhibit NLRP3 inflammasome activation and its potential usage in the treatment of inflammatory bowel diseases.
Subject(s)
Carrier Proteins/metabolism , Colitis/drug therapy , Colon/drug effects , Inflammasomes/drug effects , Inflammatory Bowel Diseases/drug therapy , Mitochondria/drug effects , Pentacyclic Triterpenes/administration & dosage , Animals , Caspase 1/metabolism , Cells, Cultured , Centella/immunology , Colitis/chemically induced , Colitis/immunology , Colon/immunology , Colon/pathology , Cytokines/metabolism , Dextran Sulfate/administration & dosage , Female , Humans , Immunosuppression Therapy , Inflammasomes/metabolism , Inflammation Mediators/metabolism , Inflammatory Bowel Diseases/immunology , Mice , Mice, Inbred C57BL , Mitochondria/metabolism , Models, Animal , NLR Family, Pyrin Domain-Containing 3 ProteinABSTRACT
S3A was a RG-I pectin isolated from Centella asiatica that contained Rha, Ara, Gal, Glc and GalA in molar ratio of 1.0:0.6:1.5:0.2:1.1 and had been found to have a backbone composed mainly of the disaccharide repeat unit, -->4)-alpha-D-GalpA-(1-->2)-alpha-L-Rhap-(1-->. Based on methylation analysis, NaIO4 oxidation, partial acid hydrolysis and lithium-treatment, the structural features were elucidated. Side chains of S3A were predominantly linked to O-4 of 1,2,4-linked alpha-L-Rhap. The side chains are comprised of arabinosyl chains, galactosyl chains, arabinogalactosyl chains and short glucosyl chains. A total of 45% Rhap in the backbone was substituted by side chains. The arabinosyl residues were mostly distributed in the arabinosyl side chains. According to the immunological results of S3A and its degraded derivatives, S3A had no immunological activity, but its derivatives had immuno-stimulating activities to some extent.
