Regulation of pro and anti-inflammatory signaling molecules in effect of Withania Somnifera root extract treated sleep deprivation induced Wistar rats.

Sleep plays an imperative role in maintaining good health. Sleep along with circadian cycle wields strong regulatory control over immunity. Sleep deprivation (SD) is a threat to health developing several immunological disorders. The medicinal plant Withania somnifera (WS) root extract is widely used for its immuno-modulatory properties. Therefore, it is of interest to assess the effect of WS root extract on pro and anti-inflammatory signalling in SD rats. 24 male Wistar rats (120-150g) were divided into 4 groups with 6 animals in each. The groups were divided such that Group I - cage control, Group II - large platform control, Group III - sleep deprived & Group IV - WS treated SD rats. RT-PCR based mRNA expression analysis of pro inflammatory (IL-1ß, IL-6, MCP-1, TNF-α) and anti-inflammatory marker (IL-10) in the cortex of control and SD rats were completed. Concurrent protein expression analysis was completed using western blot. Data was analyzed using one-way ANOVA and Duncan's multiple range test in SPSS software version 20. Data showed elevation of pro-inflammatory markers and depression of IL-10. Thus, WS down regulated the pro-inflammatory and up-regulated the anti-inflammatory molecules, which can be further considered towards the treatment of sleep deprivation induced inflammatory diseases.

A mechanistic and empirical review of antcins, a new class of phytosterols of formosan fungi origin.

Antcins are unique phytosterols isolated from A. cinnamomea and A. salmomea, which are the endemic fungus of Taiwan. A. cinnamomea has long been highly valued medicinal mushroom in Taiwan and traditionally used as a folk remedy for various human illness. Recent scientific explorations claimed that the pharmacological activities of A. cinnamomea and A. salmomonea are gone beyond their original usage. The therapeutic efficacy of these medicinal mushrooms was attributed to their high content of unique bioactive secondary metabolites, including terpenoids, benzenoids, ubiquinol derivatives, polysaccharides, lignans, nucleic acids, steroids, and maleic/succinic acid derivatives. Antcins is a group of steroids in Antrodia spp. with ergostane skeleton received much attention in Taiwan's academic circle due to their broad-spectrum of biological activities. At present, twelve antcins, i.e. antcin A, B, C, D, E, F, G, H, I, K, M, and N along with twelve derivatives/epimers (25R/S-antcin A, B, C, H, I and K) and seven analogs (methyl antcinate A, B, G, H, K, L and N) were identified. Several studies have demonstrated that antcins possessed anti-cancer, anti-inflammation, anti-oxidant, anti-diabetic, anti-aging, immunomodulation, hepatoprotection, and hypolipedimic activities. The main goal of this review is to define the chemistry, isolation, advances in production, and biological activities of antcins and their derivatives/analogs. Special attention has been given to a detail view of their biological activities in vitro and in vivo and their pharmacological potentials.

A steroid like phytochemical Antcin M is an anti-aging reagent that eliminates hyperglycemia-accelerated premature senescence in dermal fibroblasts by direct activation of Nrf2 and SIRT-1.

The present study revealed the anti-aging properties of antcin M (ANM) and elucidated the molecular mechanism underlying the effects. We found that exposure of human normal dermal fibroblasts (HNDFs) to high-glucose (HG, 30 mM) for 3 days, accelerated G0/G1 phase arrest and senescence. Indeed, co-treatment with ANM (10 µM) significantly attenuated HG-induced growth arrest and promoted cell proliferation. Further molecular analysis revealed that ANM blocked the HG-induced reduction in G1-S transition regulatory proteins such as cyclin D, cyclin E, CDK4, CDK6, CDK2 and protein retinoblastoma (pRb). In addition, treatment with ANM eliminated HG-induced reactive oxygen species (ROS) through the induction of anti-oxidant genes, HO-1 and NQO-1 via transcriptional activation of Nrf2. Moreover, treatment with ANM abolished HG-induced SIPS as evidenced by reduced senescence-associated ß-galactosidase (SA-ß-gal) activity. This effect was further confirmed by reduction in senescence-associated marker proteins including, p21CIP1, p16INK4A, and p53/FoxO1 acetylation. Also, the HG-induced decline in aging-related marker protein SMP30 was rescued by ANM. Furthermore, treatment with ANM increased SIRT-1 expression, and prevented SIRT-1 depletion. This protection was consistent with inhibition of SIRT-1 phosphorylation at Ser47 followed by blocking its upstream kinases, p38 MAPK and JNK/SAPK. Further analysis revealed that ANM partially protected HG-induced senescence in SIRT-1 silenced cells. A similar effect was also observed in Nrf2 silenced cells. However, a complete loss of protection was observed in both Nrf2 and SIRT-1 knockdown cells suggesting that both induction of Nrf2-mediated anti-oxidant defense and SIRT-1-mediated deacetylation activity contribute to the anti-aging properties of ANM in vitro. Result of in vivo studies shows that ANM-treated C. elegens exhibits an increased survival rate during HG-induced oxidative stress insult. Furthermore, ANM significantly extended the life span of C. elegans. Taken together, our results suggest the potential application of ANM in age-related diseases or as a preventive reagent against aging process.

2,3,5-Trimethoxy-4-cresol, an anti-metastatic constituent from the solid-state cultured mycelium of Antrodia cinnamomea and its mechanism.

Antrodia cinnamomea is a valuable and unique edible fungus originating from the forests of Taiwan. In this study, an anti-metastatic compound, 2,3,5-trimethoxy-4-cresol (TMC), was isolated from the solid-state cultured mycelium of A. cinnamomea. According to the results obtained from cell wound healing, cell migration and invasion assays, TMC effectively suppressed movement, migration and invasion of lung cancer cells at the dosage of 5-40 µM, which was non-toxic to A549 cells. In addition, TMC reduced protein expression of Akt, MMP-2 and MMP-9 and enhanced E-cadherin and TIMP-1 protein expression, which are known to regulate cell adhesion, migration and invasion. Taken together, TMC effectively suppresses movement, migration and invasion of lung cancer cells, and achieves an anti-cancer metastasis effect.

Investigation on antidiarrhoeal activity of Aristolochia indica Linn. Root extracts in mice.

BACKGROUND: The present study aimed at investigating the effect of ethanolic extract (EtAI), and aqueous extract (AqAI) of Aristolochia indica Linn roots on castor oil-induced diarrhoea and study on small intestinal transit. Phytochemical analysis of extracts was performed as per standard procedure. MATERIALS AND METHODS: The oral toxicity study using Swiss albino mice was performed in accordance with OECD guidelines. The EtAI and AqAI extracts of Aristolochia indica Linn were studied for antidiarrhoeal property using castor oil-induced diarrhoeal model and charcoal-induced gastrointestinal motility test in Swiss albino mice. RESULTS: Among the tested doses of 200 and 400 mg/kg body weight, the extracts reduced the frequency and severity of diarrhoea in test animals throughout the study period. At the same doses, the extract delayed the intestinal transit of charcoal meal in test animals as compared to the control and the results were statistically significant. CONCLUSION: Experimental findings showed that ethanol extract of Aristolochia indica Linn root possess significant antidiarrheal activity and may be a potent source of anti-diarrhoeal drug in future.

Antrodia salmonea inhibits TNF-α-induced angiogenesis and atherogenesis in human endothelial cells through the down-regulation of NF-κB and up-regulation of Nrf2 signaling pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Antrodia salmonea (AS) is known as a traditional Chinese medicine, but very few biological activities have been reported. MATERIALS AND METHODS: The present study was aimed to investigate the anti-angiogenic and anti-atherosclerotic potential of the fermented culture broth of AS against tumor necrosis factor-α (TNF-α)-stimulated human endothelial (EA.hy 926) cells. RESULTS: The non-cytotoxic concentrations of AS significantly inhibited TNF-α-induced migration/invasion and capillary-like tube formation in EA.hy 926 cells. Furthermore, AS suppressed TNF-α-induced activity and expression of matrix metalloproteinase-9 (MMP-9), and cell-surface expression of intercellular adhesion molecule-1 (ICAM-1), which was associated with abridged adhesion of U937 leukocytes to endothelial cells. Moreover, AS significantly down-regulated TNF-α-induced nuclear translocation and transcriptional activation of nuclear factor κB (NF-κB) followed by suppression of I-κB degradation and phosphorylation of I-κB kinase-α (IKKα). Notably, the protective effect of AS was directly correlated with the increased expression of hemeoxygenase-1 (HO-1) and γ-glutamylcysteine synthetase (γ-GCLC), which was reasoned by nuclear translocation and transactivation of NF-E2 related factor-2 (Nrf2)/antioxidant response element (ARE). Furthermore, HO-1 knockdown by HO-1-specific shRNA diminished the protective effects of AS on TNF-α-stimulated invasion, tube formation, and U937 adhesion in EA.hy 926 cells. CONCLUSIONS: Taken together, these results suggest that Antrodia salmonea may be useful for the prevention of angiogenesis and atherosclerosis.

The Gastroprotective Role of Acanthus ilicifolius - A Study to Unravel the Underlying Mechanism of Anti-Ulcer Activity.

Acanthus ilicifolius (Acanthaceae), a mangrove medicinal plant, is widely used by the local inhabitants of the Sundarbans (India) to treat a variety of diseases. As a part of our continued search for novel bioactive products from mangrove medicinal plants, we were able to document the anti-inflammatory effects of this plant. In the present study, we have performed a detailed evaluation of the gastroprotective activity of the methanolic extract of Acanthus ilicifolius using different models of gastric ulceration. Unlike the conventional non-steroidal anti-inflammatory drugs, a methanolic extract of Acanthus ilicifolius leaves (MEAL) possessing significant anti-inflammatory properties, as revealed from our previous studies displayed in rats in dosages of 200 mg and 400 mg/kg BW after intraperitoneal administration, showed significant protective activity (anti-ulcer activity) against the gastric lesions induced by aspirin, indomethacin, stress, ethanol, and pylorus ligation. In pylorus-ligated rats, administration of Methanolic extract of Acanthus ilicifolius leaves (MEAL) significantly decreased gastric volume, acidity, and peptic activity. Moreover, pre-treatment with MEAL significantly restored the levels of reduced glutathione (GSH) and the antioxidant enzyme superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX), along with significant inhibition of both lipid peroxidation and myeloperoxidase (MPO) activity in pylorus-ligated animals. Ulceration induced with ethanol was significantly inhibited with MEAL, and the extract also resulted in the reduction of both lipid peroxidation and myeloperoxidase activity. Furthermore, in this experimental model, administration of MEAL improved the activities of SOD, CAT, GSH, and GPX. A similar pattern of action was also noticed in cold-restraint stress-induced (CRS) ulceration, where MEAL pre-treatment inhibited CRS-induced ulceration, improved the status of antioxidant enzymes, and also reduced the level of lipid peroxides. These results suggest that extracts of the leaves of Acanthus ilicifolius may exhibit anti-ulcer activities additional to the anti-inflammatory properties.

In vitro and in vivo activity of gallic acid and Toona sinensis leaf extracts against HL-60 human premyelocytic leukemia.

Toona sinensis is one of the most popular vegetarian cuisines in Taiwan and it has been shown to induce apoptosis in cultured human premyelocytic leukemia (HL-60) cells. In the present study, we examined the effects of T. sinensis leaf extracts (TS extracts) on tumor regression using in vitro cell culture and an in vivo athymic nude mice model. We found that TS extracts (10-75 µg/mL) arrested HL-60 cells at the G1-S transition phase through the reductions of Cyclin D1, CDK4, Cyclin E, CDK2, and Cyclin A, and induction of CDK inhibitor p27KIP levels. Furthermore, VEGF expression and release was significantly inhibited by TS extracts. Notably, TS extracts treatment was effective in terms of delaying tumor incidence in the nude mice inoculated with HL-60 cells as well as reducing the tumor burden. Histological analysis confirmed that TS extracts significantly modulated tumor progression in xenograft tumor. Furthermore, a similar pattern of results were observed from gallic acid (5 and 10 µg/mL), a major compound in TS, caused G1 arrest through regulations of cell-cycle regulatory proteins. Our data suggest that T. sinensis exerts antiproliferative effects on HL-60 cells in vitro and in vivo due mainly to the presence of gallic acid.

Antioxidant and anti-inflammatory potential of hesperetin metabolites obtained from hesperetin-administered rat serum: an ex vivo approach.

In recent years much attention has been focused on the pharmaceutical relevance of bioflavonoids, especially hesperidin and its aglycon hesperetin in terms of their antioxidant and anti-inflammatory actions. However, the bioactivity of their metabolites, the real molecules in vivo hesperetin glucuronides/sulfates produced after ingestion, has been poorly understood. Thus, the study using an ex vivo approach is aimed to compare the antioxidant and anti-inflammatory activities of hesperidin/hesperetin or hesperetin metabolites derived from hesperetin-administered rat serum. We found that hesperetin metabolites (2.5-20 µM) showed higher antioxidant activity against various oxidative systems, including superoxide anion scavenging, reducing power, and metal chelating effects, than that of hesperidin or hesperetin. The data also showed that pretreatment of hesperetin metabolites (1-10 µM) within the range of physiological concentrations, compared to hesperetin, significantly inhibited nitric oxide (NO) and prostaglandin E(2) (PGE(2)) production, as evidenced by the inhibition of their precursors, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) protein levels without appreciable cytotoxicity on LPS-activated RAW264.7 macrophages or A7r5 smooth muscle cells. Concomitantly, hesperetin metabolites dose-dependently inhibited LPS-induced intracellular reactive oxygen species (ROS). Furthermore, hesperetin metabolites significantly downregulate LPS-induced nuclear factor-κB (NF-κB) activation followed by the suppression of inhibitor-κB (I-κB) degradation and phosphorylation of c-Jun N-terminal kinase1/2 (JNK1/2) and p38 MAPKs after challenge with LPS. Hesperetin metabolites ex vivo showed potent antioxidant and anti-inflammatory activity in comparison with hesperidin/hesperetin.

Lucidone inhibits iNOS and COX-2 expression in LPS-induced RAW 264.7 murine macrophage cells via NF-kappaB and MAPKs signaling pathways.

The anti-inflammatory mechanism of lucidone isolated from the fruits of Lindera erythrocarpa Makino was investigated. Our data indicate that lucidone significantly inhibits the production of NO and PGE(2) autacoids in LPS-induced RAW 264.7 murine macrophage cells. Moreover, it also notably decreased the secretion of tumor necrosis factor-alpha (TNF-alpha). Consistent with these observations, the mRNA and protein expression levels of iNOS and COX-2 were also inhibited by lucidone in a dose-dependent manner. Lucidone also reduced the translocation of NF-kappaB induced by LPS, which is associated with the prevention of the degradation of I-kappaB, and subsequently decreased p65/p50 protein levels in the nucleus. Lucidone also inhibited NF-kappaB activation by impairing the binding of NF-kappaB to its cis-acting element. In addition, lucidone inhibited JNK and p38MAPKs signals, which are the most significant signals involved in NO, PGE(2) and TNF-alpha production; NF-kappaB/AP-1 activation was also inhibited by lucidone. Taken together, the anti-inflammatory activity of lucidone might be caused by the inhibition of iNOS and COX-2 expressions through downregulation of NF-kappaB and AP-1 binding.

Anti-inflammatory activity of Acanthus ilicifolius.

Acanthus ilicifolius Linn, is a perennial herb (Acanthaceae) widely found in the Sundarban mangroves and is popularly used for its wound healing effects. In the present study an attempt was made to evaluate the anti-inflammatory activity of the Acanthus ilicifolius leaves. The methanolic fraction of Acanthus ilicifolius leaf extract produced significant inhibition of rat paw oedema, when administered both prior to and after carrageenan administration, in a manner similar to BW755C a synthetic cyclooxygenase (COX) and lipoxygenase (LOX) inhibitor. The extract decreased protein exudation and leukocyte migration in the peritoneal fluid, thereby indicating its effectiveness towards inhibiting peritoneal inflammation. It also produced significant inhibition of COX (1 and 2) and 5-LOX activity. Preincubation of the extract inhibited the production of proinflammatory cytokines (TNFalpha and IL-6) in lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells (PBMCs). The methanolic fraction of the extract was also found to possess significant free radical (DPPH, ABTS, superoxide and hydroxyl radical) scavenging activity. The extract on intraperitoneal administration augmented the endogenous antioxidant status, as evident from the significant increase of ferric reducing ability of plasma (FRAP) and total peroxyl radical trapping activity of plasma (TRAP).