Inhibitory effects of Angelica sinensis ethyl acetate extract and major compounds on NF-κB trans-activation activity and LPS-induced inflammation

Aim of the study

Previous study showed that the ethyl acetate (EtOAc) fraction from Angelica sinensis (Oliv.) Diels (Apiaceae) (AS) inhibited nitric oxide (NO) and prostaglandin E₂ secretions in vitro. This study was to evaluate anti-inflammatory activity of AS EtOAc extract and its major compounds in vivo and in vitro.

Materials and methods

NF-κB luciferase activity and pro-inflammatory cytokine secretions from lipopolysaccharide (LPS) plus interferon (IFN)-γ-stimulated RAW 264.7 cells pre-treated with EtOAc extract or compounds were analyzed. For further in vivo study, BALB/c mice were tube-fed with 1.56 (AS1 group), 6.25 (AS2 group) mg/kg body weight/day in 100 μl soybean oil, while the control and PDTC (pyrrolidine dithiocarbamate, an anti-inflammatory agent) groups were tube-fed with 100 μl soybean oil/day only. After 1 week of tube-feeding, the PDTC group was injected with 50 mg/kg BW PDTC and 1 h later, all of the mice were injected with 15 mg/kg BW LPS. The pro-inflammatory cytokine levels and lifespan of LPS-challenged mice were determined.

Results

The results showed that AS EtOAc extract significantly inhibited NF-κB luciferase activity and TNF-α, IL-6, macrophage inflammatory protein-2 (MIP-2) and NO secretions from LPS/IFN-γ-stimulated RAW 264.7 cells. The AS1 and PDTC groups, but not AS2, had significantly higher survival rate than the control group. This was characterized by the inhibition of the serum TNF-α and IL-12p40 levels after LPS injection (p < 0.05). The major compounds of AS, ferulic acid and Z-ligustilide, also significantly decreased NF-κB luciferase activity, which may contribute to the anti-inflammatory activity of AS.

Conclusions

Low dose of AS EtOAc extract that inhibits the production of inflammatory mediators alleviates acute inflammatory hazards and protect mice from endotoxic shock.     

Total flavonoids of Mosla scabra leaves attenuates lipopolysaccharide-induced acute lung injury via down-regulation of inflammatory signaling in mice

Ethnopharmacological relevance

Mosla scabra (Thunb.) C.Y. Wu, belonging to the Labiatae family, is a tomentose and aromatic plant, which is widely used as an antipyretic and antiviral drug for pulmonary diseases and famous for its efficiency in treating colds, fever, pneumonia and chronic bronchitis. To investigate therapeutic effects and possible mechanism of Mosla scabra flavonoids (MF) on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice.

Materials and methods

Mice were orally administrated with MF once (30 mg/kg or 90 mg/kg) 1 h before LPS challenge. Lung specimens and the bronchoalveolar lavage fluid (BALF) were isolated for histopathological examinations and biochemical analyses 6 h after LPS challenge.

Results

Pretreatment with MF could decrease significantly lung wet-to-dry weight (W/D) ratio, lower myeloperoxidase (MPO) activity and total protein concentrations in the BALF, reduce serum levels of NO, TNF-α, IL-1β and IL-6 in ALI model. Additionally, MF attenuated lung histopathological changes and significantly inhibited the phosphorylation of p38 MAPK and translocation of NF-κB p65.

Conclusions

These results showed MF significantly attenuate LPS-induced acute lung injury and production of inflammatory mediators via inhibiting MAPK and NF-κB activation, indicating it as a potential therapeutic agent for ALI.     

Ethanol extract of Alismatis Rhizoma reduces acute lung inflammation by suppressing NF-κB and activating Nrf2

Ethnopharmacological relevance

The tuber of Alisma orientale Juzepzuk, a medicinal herb that has been used for the treatment of various disorders in Korea, has an anti-inflammatory effect. Here, we investigated a possible underlying mechanism and a protective effect on acute lung injury (ALI).

Materials and methods

Alisma orientale tuber was extracted in 80% ethanol and dried. The powder of the ethanol extract of Alisma orientale tuber (EEAO) was dissolved in PBS. The effect of EEAO on NF-κB and Nrf2 activities was analyzed with RAW 264.7 cells. The effect of EEAO on lung inflammation was determined by histologic and molecular biological analyses of the lung tissue of C57BL/6 mice that were gavaged once a day with 0.3 or 1.2 g/kg of EEAO for 14 days, prior to an intranasal administration of LPS (0.01 g/kg) for inducing ALI.

Results

EEAO pre-treatment of RAW 264.7 cells suppressed NF-κB activity and the expression of its dependent genes including COX-2, IL-1β and iNOS. Similar treatment enhanced Nrf2 activity and the expression of Nrf2-regulated genes including NQO-1, HO-1 and GCLC. LPS instillation induced acute neutrophilic lung inflammation, which was significantly suppressed by pre-treatment with EEAO. Analysis of the lungs revealed that EEAO pre-treatment induced the expression of Nrf2-regulated genes, with concomitant down-regulation of inflammatory gene expression.

Conclusions

EEAO attenuated lung inflammation in LPS-induced ALI mice, which was associated with differential regulation of NF-κB and Nrf2 activities. We suggest that EEAO can be developed as a potential therapeutics for the treatment of ALI.     

Anti-inflammatory action of hydroalcoholic extract,dichloromethane fraction and steroid α-spinasterol from Polygala sabulosa in LPS-induced peritonitis in mice

Ethnopharmacological relevance

Polygala sabulosa A. W. Bennett is a small herb popularly known as “timutu-pinheirinho” that is widely distributed in southern Brazil and that is used to treat disorders of the bowel and kidney and as a topical anesthetic and expectorant in folk medicine. This study was designed to investigate the anti-inflammatory properties of the hydroalcoholic extract (HEPs), CH₂Cl₂ fraction and the steroid α-spinasterol obtained from the aerial parts of Polygala sabulosa in a model of acute inflammation induced by intraperitoneal injection of bacterial lipopolysaccharide in mice.

Materials and methods

The anti-inflammatory effect of HEPs (3–300 mg/kg, i.g.), CH₂Cl₂ fraction (0.003–30 mg/kg, i.g.) and steroid α-spinasterol (0.001–1 mg/kg, i.p. or 1–10 mg/kg, i.g.), were evaluated in mice subjected to the acute inflammation caused by intraperitoneal (i.p.) injection of lipopolysaccharide (LPS, 0.02 µg/kg). The anti-inflammatory activity of the HEPs, CH₂Cl₂ fraction and steroid were assessed by determining the total numbers of leukocytes and differential cell counts (neutrophils and mononuclear cells) and levels of pro-inflammatory (IL-1β, TNF-α, IL-6) or anti-inflammatory (IL-10) cytokines in peritoneal fluid.

Results

The administration of HEPs (3–300 mg/kg, i.g.) completely inhibited inflammatory cell infiltration (300 mg/kg, i.g.) and it reduced TNF-α (100–300 mg/kg) and IL-1β (100 mg/kg) levels in LPS-injected mice. Furthermore, the administration of CH₂Cl₂ fraction (0.003–30 mg/kg, i.g.) or α-spinasterol (0.001–10 mg/kg, by i.p. or i.g.) significantly reduces inflammatory cell infiltration in LPS-injected mice. Moreover, dexamethasone (0.5 mg/kg, i.p., used as a positive control) inhibited inflammatory cell infiltration and reduced the levels of TNF-α, IL-1β and IL-6 in LPS-injected mice.

Conclusions

Taken together, these results provide the first experimental evidence demonstrating that HEPs have significant anti-inflammatory effects on LPS-induced inflammation. These effects appear to be, at least in part, due to the presence of α-spinasterol. These findings support the widespread use of Polygala sabulosa in popular medicine and demonstrate that this plant has therapeutic potential for the development of phytomedicines with anti-inflammatory properties.     

Safety evaluation of main alkaloids from Rhizoma Coptidis

Ethnopharmacological relevance

Rhizoma Coptidis (RC), a widely used traditional Chinese medicine, has been used for the treatment of heat-clearing and detoxifying, but there is very little information on its safety.

Aim of the study

To provide information on the safety of RC, we evaluated the toxicity of the crude RC and RC alkaloids (berberine, coptisine, palmatine and epiberberine) including cytotoxicity, acute toxicity in mice and sub-chronic toxicity in rats.

Materials and methods

The cytotoxicity of RC alkaloids was tested in HepG2 and 3T3-L1 cells by the MTT assay. The acute toxicity of RC alkaloids was tested in mice and the mortality was calculated at the end of experiment. For sub-chronic toxicity study, the rats were treated with the RC alkaloids at a dose of 156 mg/kg/day and RC at a dose of 521 mg/kg/day for 90 days. Mortality, clinical signs, body weight changes, organ weights, urinalysis and hematological parameters, gross necropsy and histopathology were monitored during the study period.

Results

The cell assay indicates that the IC₅₀ values of berberine, coptisine, palmatine and epiberberine in HepG2 cells were 48.17, 64.81, 112.80 and 120.58 μg/mL, which in 3T3-L1 cells were 41.76, 56.48, 84.32 and 104.18 μg/mL, respectively. In the acute toxicity assay, the LD₅₀ values of four alkaloids were 713.57, 852.12, 1533.68 and 1360 mg/kg, respectively. However, in the sub-chronic toxicity study, no mortality and morbidity were observed which could be related to RC alkaloids and RC treatment. Besides, there was no abnormality in clinical signs, body weights, organ weights, urinalysis, hematological parameters, gross necropsy and histopathology in any of the animals after the oral administration of RC alkaloids and RC.

Conclusions

Taking these results together, we came to the conclusion that the toxicity of berberine is the maximum and palmatine is the minimal in four RC alkaloids. The currently recommended doses of RC alkaloids and RC consumed are relatively safe.     

Inhibition of airway inflammation by the roots of Angelica decursiva and its constituent,columbianadin

Ethnopharmacological relevance

The roots of Angelica decursiva Fr. Et Sav (Umbelliferae) have been frequently used in traditional medicine as anti-inflammatory, antitussive, analgesic agents and expectorant, especially for treating cough, asthma, bronchitis and upper respiratory tract infections. To establish the scientific rationale for the clinical use of Angelica decursiva and to identify new agents for treating inflammatory lung disorders, pharmacological evaluation of the roots of Angelica decursiva and the isolated constituents was performed.

Methods

In vitro study was carried out using two lung cells, lung epithelial cells (A549) and alveolar macrophages (MH-S). The inflammatory markers such as IL-6 and nitric oxide (NO) for each cell line were examined. For in vivo study, a mouse model of lipopolysaccharide (LPS)-induced acute lung injury was used and the effects on lung inflammation were established by measuring the cell numbers in bronchoalveolar lavage fluid (BALF) and by histological observation.

Results

Water and 70% ethanol extracts of the roots of Angelica decursiva showed considerable inhibitory activity against LPS-induced lung inflammation in mice following oral administration at a dose of 400 mg/kg. Five coumarin derivatives including columbianadin, umbelliferone, umbelliferone 6-carboxylic acid, nodakenin and nodakenetin were isolated. Among the isolated compounds, columbianadin was found to possess strong inhibitory activity against the inflammatory response of IL-1β-treated A549 cells and LPS-treated MH-S cells. Columbianadin was found to inhibit NO production by down-regulation of inducible NO synthase. Moreover, columbianadin was also proved to possess significant inhibitory activity against LPS-induced lung inflammation following oral administration at a dose of 20–60 mg/kg.

Conclusions

The roots of Angelica decursiva were proved to be effective in the treatment of lung inflammation. Columbianadin can be a potential new agent for treating inflammatory lung disorders.     

Effects of Hominis placenta on LPS-induced cell toxicity in BV2 microglial cells

Ethonopharmacological relevance

Hominis placenta (HP) dried placenta extracted from pregnant women after delivery has been widely used to treat chronic inflammatory diseases. HP has been reported to be effective to alleviate the arthritic symptoms by modulating the expression of inflammatory factors in adjuvant-induced arthritis rats. However, the mechanism of action of HP is unknown. Neuroinflammation has been implicated in the pathogenesis of several neurodegenerative disease, including Alzheimer's disease (AD), Parkinson's disease (PD) and amyotropic lateral sclerosis (ALS). Activated microglia produce large amounts of toxic soluble factors, which can be responsible for the neurodegenerative disease. Chronic microglial activation leads to neuroinflammation, which contributes to neuronal dysfunction, injury and loss in these diseases. Lipopolysaccharide (LPS) is widely used for neuroinflammation caused by microglial activation of immune cells in the central nervous system (CNS) and subsequent release of inflammatory or neurotoxic factors. In the present study, we investigated the effects and signaling pathway of HP in the LPS induced BV2 microglial cells.

Materials and method

BV2 microglial cells were pretreated with 50 μM HP for 2 h prior to 2 μg/ml LPS for 15 min. Cell viability was determined by MTT assay. The level of protein expression was analyzed by western blot. Immunofluorescence was performed with an anti-COX2 antibody in BV2 cells.

Results

HP decreased LPS-induced microglial cell death by 24% and inhibited LPS-induced activation of c-Jun N-terminal kinase (JNK) by 23% and p42/44MAP kinase (ERK) by 34% treatment of LPS. In addition, HP attenuated LPS-induced pro-inflammatory proteins such as iNOS and COX2 in microglial cells 34% and 28% respectively.

Conclusions

Our data shows that HP has a protective role against LPS stimulation through inhibition of MAPK signaling and suppression of inflammation caused by neurotoxin including LPS. These findings suggest that HP could be a potential therapeutic agent of neurodegenerative diseases which accompanied with microglial activation.     

The anti-inflammatory effects of Caragana tangutica ethyl acetate extract

Ethnopharmacological relevance

Caragana tangutica KOM has been used to treat arthritis, wounds, fever and other disease conditions in traditional Chinese medicine (TMC). To support the application of the plant in traditional Chinese medicine by investigating the anti-inflammatory effects of the ethyl acetate extract of Caragana tangutica.

Materials and methods

The anti-inflammatory activity was evaluated by animal models including xylene-induced ear edema in mice, carrageenan-induced paw edema in rats, acetic acid induced writhing in mice and LPS-induced acute lung injury (ALI). The anti-inflammatory mechanism was evaluated by detecting prostaglandin E₂ and immunohistochemistry expression of cyclooxygenase-2 (COX-2) using an EIA assay kit and immunohistochemistry, respectively.

Results

The results showed that the xylene-induced ear edema in mice was significantly reduced by the ethyl acetate extract at dosages of 100, 200 and 400 mg/kg, and the carrageenan-induced paw edema in rats was monitored to be reduced by the ethyl acetate extract 3 h after carrageenan injection. The ethyl acetate extract was also found to reduce the inflammation pain of acetic acid-induced writhing model in a dose-dependent manner and cause reduction of the ALI in mice through the inhibition of the release of PGE₂ and the LPS-induced COX-2 expression in the lung.

Conclusion

Our study demonstrates that the ethyl acetate extract of the plant can help to reduce inflammations by inhibiting the expression of COX-2.     

Preventive and therapeutic effects of Danhong injection on lipopolysaccharide induced acute lung injury in mice

Ethnopharmacological relevance

Danhong injection (DHI), a Chinese Materia Medica standardized product extracted from Radix et Rhizoma Salviae Miltiorrhizae (Salvia miltiorrhiza Bge., Labiatae, Danshen in Chinese) and Flos Carthami (Carthamus tinctorius L., Compositae, Honghua in Chinese), has been reported to have anti-inflammatory, anti-oxidative and anti-fibrinolytic properties, which is used extensively for the treatment of cardiovascular diseases in clinic.

Aim of this study

The present study aimed to investigate the preventive and therapeutic effects of DHI on lipopolysaccharide (LPS) induced acute lung injury (ALI) in mice.

Materials and methods

Lung injury was induced by intranasal instillation with 10 μg LPS. Mice were randomly divided into four groups:Control group; LPS group; LPS+5 ml/kg DHI group and LPS+10 ml/kg DHI group. The effects of DHI on LPS-induced neutrophils influx, inflammatory cytokines release, protein leakage, myeloperoxidase (MPO) and superoxide dismutase (SOD) activities, malondialdehyde (MDA) level were examined. In addition, the NF-κB activation in lung tissues was detected by Western blot.

Results

In LPS challenged mice, DHI significantly reduced the infiltration of activated neutrophils and decreased the levels of TNF-α and IL-6 in bronchoalveolar lavage fluid (BALF). DHI also inhibited protein extravasation in BALF, attenuated edema and the pathological changes in the lung. In addition, DHI markedly prevented LPS-induced elevation of MDA and MPO levels, as well as reduction of SOD activity. Further study demonstrated that DHI effectively inhibited the NF-κB activation in lung tissues.

Conclusion

DHI has been demonstrated to protect mice from LPS induced acute lung injury by its anti-inflammatory and anti-oxidant activities.     

Carthamus tinctorius L. prevents LPS-induced TNFα signaling activation and cell apoptosis through JNK1/2–NFκB pathway inhibition in H9c2 cardiomyoblast cells

Aim of the study

Severe and potentially fatal hypotension and cardiac contractile dysfunction are common symptoms in patients with sepsis. In our previous study, we found that estradiol and estrogen-receptor α have cardio-protective effects in myocardial cells exposed to LPS. Estradiol supplementation has been shown to induce breast and cervical cancers. Flos Carthami, the flower of Carthamus tinctorius L. (Compositae) is an important traditional Chinese medicine used for the treatment of heart disease and inflammation, and therefore might be a potential alternative to Estradiol in the prevention of heart damage. This study investigated the effect of Flos Carthami (FC_EtOH) ethanolic extract on LPS-induced apoptosis in H9c2 cardiomyoblast cells.

Materials and methods

H9c2 cells induced apoptosis with LPS administration (1 μg/mL). H9c2 cells were divided into five groups: Control, LPS (1 μg/mL), and three FC_EtOH (31.25, 62.5,and 125 μg/mL). We detected apoptosis using MTT, LDH, TUNEL assay. JC-1 staining and Western blot were used to detect pro-apoptosis proteins, anti-apoptosis proteins, MAPK proteins (JNK, ERK, and P38), and NFκB expression.

Results

FC_EtOH (62.5 μg/mL) inhibited LPS-induced apoptosis by suppressing JNK1/2 activity, which resulted in the reduction of both IκB degradation and NFκB activation. In addition, FC_EtOH led to the activation of anti-apoptotic proteins, Bcl-2 and Bcl-xL, the stabilization of the mitochondria membrane and the down-regulation of extrinsic and intrinsic pro-apoptotic proteins, such as TNFα, active caspase-8, t-Bid, Bax, active caspases-9, and -3.

Conclusions

Carthamus tinctorius L. possesses the ability to suppress JNK activity and inhibit LPS-induced TNFα activation and apoptosis in H9c2 cardiomyoblast cells. Carthamus tinctorius L could potentially serve as a cardio-protective agent against LPS-induced apoptosis.     

The effect of the aqueous extract of Helietta parvifolia A. Gray (Rutaceae) stem bark on carrageenan-induced paw oedema and granuloma tissue formation in mice

Aims of study

Despite the ethnopharmacological relevance of Helietta parvifolia A. Gray (Rutaceae) in Mexico, we found no significant pharmacological studies of this plant in the scientific literature. The aim of the present study was to establish the anti-inflammatory effect of an aqueous extract of the stem bark of Helietta parvifolia in mice.

Materials and methods

The anti-inflammatory activity of the aqueous extract of the stem bark of Helietta parvifolia was evaluated using carrageenan-induced paw oedema in mice, and the cotton pellet granuloma method.

Results

An extract dose ranging from 20 to 80 mg/kg p.o. showed a non-significant effect over the initial phase of carrageenan-induced oedema. However, it showed a significant inhibition of oedema after 3 h, which can be related to the inhibition of the release of kinin-like substances. An ID₅₀ value of 47.4 mg/kg was obtained for the plant extract. The extract also suppressed granulomatous tissue formation during chronic inflammation. The inhibitory values were 19.2, and 22.2, corresponding to 40 and 80 mg/kg doses of extract respectively.

Conclusions

Aqueous extract showed a statistically significant anti-inflammatory effect in mice during the late phase of acute inflammation and during chronic inflammation. However, the exact mechanism(s) of anti-inflammatory effects of Helietta parvifolia observed in this study remains unclear.     

Antiviral effects of modified Dingchuan decoction against respiratory syncytial virus infection in vitro and in an immunosuppressive mouse model

Ethnopharmacological relevance

Modified Dingchuan decoction (MDD) is used in traditional Chinese medicine for the treatment of cough, chronic bronchitis, asthma and viral pneumonia.

Aim of the study

To investigate antiviral potentials of MDD in respiratory syncytial virus (RSV) infected mice.

Materials and methods

MDD and each component were evaluated for antiviral efficacy against RSV in vitro in cell culture. Mice were were treated with cyclophosphamide and infected with RSV. Then, treatments with MDD at doses of 1.75 g/kg, 3.5 g/kg and 7.0 g/kg, respectively, were oral administrated daily for 5 days after challenge. The levels of Eotaxin, IL-4 and IFN-γ in serum and lung tissue were detected by ELISA, viral loads in lung tissues were detected by RFQ-PCR while expressions of NF-κB and TLR4 mRNA were also detected by RFQ-PCR.

Results

A selective index of >36.8 (2.5 times greater than that observed for ribavirin) was determined in the in vitro studies for this herbal medicine. MDD exhibited significant antiviral and anti-inflammatory effects on decreasing levels of Eotaxin, IL-4 and IFN-γ in serum and lung tissue, inhibiting pneumonia, decreasing lung viral loads and reversaling RSV-induced inflammation through down-regulation of TLR4 and NF-κB mRNA expression in the lung tissue of RSV-infected mice.

Conclusions

MDD could exhibit antiviral and anti-inflammatory effects on RSV-infected mice as a suppressor of Eotaxin, IL-4 and IFN-γ. These effects appeared to be mediated by inhibitions of TLR4 and NF-κB activation. Therefore, MDD could provide an effective therapeutic approach for RSV and its subsequent viral bronchitis.     

Taraxacum officinale protects against lipopolysaccharide-induced acute lung injury in mice

Aim of the study

Taraxacum officinale has been frequently used as a remedy for inflammatory diseases. In the present study, we investigated the in vivo protective effect of Taraxacum officinale on acute lung injury (ALI) induced by lipopolysaccharide (LPS) in mice.

Materials and methods

Taraxacum officinale at 2.5, 5 and 10 mg/kg was orally administered once per day for 5 days consecutively, followed by 500 μg/kg LPS was instilled intranasally. The lung wet/dry weight (W/D) ratio, protein concentration and the number of inflammatory cells in bronchoalveolar lavage fluid (BALF) were determined. Superoxidase dismutase (SOD) and myeloperoxidase (MPO) activities, and histological change in the lungs were examined. The levels of inflammatory cytokine tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the BALF were measured using ELISA.

Results

We found that Taraxacum officinale decreased the lung W/D ratio, protein concentration and the number of neutrophils in the BALF at 24 h after LPS challenge. Taraxacum officinale decreased LPS-induced MPO activity and increased SOD activity in the lungs. In addition, histopathological examination indicated that Taraxacum officinale attenuated tissue injury of the lungs in LPS-induced ALI. Furthermore, Taraxacum officinale also inhibited the production of inflammatory cytokines TNF-α and IL-6 in the BALF at 6 h after LPS challenge in a dose-dependent manner.

Conclusions

These results suggest that Taraxacum officinale protects against LPS-induced ALI in mice.     

Antipyretic and anti-inflammatory properties of the ethanolic extract,dichloromethane fraction and costunolide from Magnolia ovata (Magnoliaceae)

Aim of the study

Magnolia ovata (A.St.-Hil.) Spreng (formerly Talauma ovata), known as “pinha-do-brejo” or “baguaçu”, is a large tree widely distributed in Brazil. Its trunk bark has been used in folk medicine against fever. However, no data have been published to support the antipyretic ethnopharmacological use. This study investigated the antipyretic and anti-inflammatory effects of the ethanolic extract (EEMO), dichloromethane fraction (DCM), and the isolated compound costunolide.

Materials and methods

The antipyretic and anti-inflammatory activities were evaluated in experimental models of fever and inflammation in mice.

Results

The oral administration of EEMO, DCM and costunolide inhibited carrageenan (Cg)-induced paw oedema (ID₅₀ 72.35 (38.64–135.46) mg/kg, 5.8 (2.41–14.04) mg/kg and 0.18 (0.12–0.27) mg/kg, respectively) and was effective in abolishing lipopolysaccharide (LPS)-induced fever (30 mg/kg, 4.5 mg/kg and 0.15 mg/kg, respectively). EEMO was also effective in reducing cell migration in the pleurisy model. Intraplantar injection of costunolide also reduced the paw oedema, myeloperoxidase and N-acetyl-glucosaminidase activity induced by Cg in mice.

Conclusions

Collectively, these results show, for the first time, that extracts obtained from Magnolia ovata possess antipyretic and anti-inflammatory properties, and costunolide appears to be the compound responsible for these effects.     

Investigation of antihyperglycaemic activity of aqueous and petroleum ether extract of stem bark of Pongamia pinnata on serum glucose level in diabetic mice

Aim of the study

The aim of the study was to evaluate the antihyperglycaemic activity of aqueous (PPSB-AQE) and petroleum ether (PPSB-PEE) extract of stem bark Pongamia pinnata in alloxan induced diabetic mice.

Materials and methods

Diabetes was induced in mice by alloxan (80 mg/kg, i.v.). After acute and subacute treatment serum glucose was determined. OGTT was performed in PPSB-PEE pretreated animals.

Results

PPSB-PEE (25, 50, 100, 200 and 400 mg/kg) showed significant reduction in serum glucose level in acute and subacute studies. The antihyperglycaemic effects of PPSBPE (100, 200 and 400 mg/kg) showed onset at 2 h and peak effect at 6 h and the effect was sustained until 24th h with 400 mg/kg. In subacute study, antihyperglycaemic effect was observed on 21st day. In PPSBPE treated mice the body weight was not reduced in contrast to that in vehicle group. In OGTT, increased glucose utilization was observed.

Conclusions

It is concluded that PPSB-PEE but not PPSB-AQE showed antihyperglycaemic activity.     

The mechanism of hepatoprotective effect of sesquiterpene rich fraction from Cichorum glandulosum Boiss. et Huet on immune reaction-induced liver injury in mice

Ethnopharmacological relevance

Cichorum glandulosum Boiss. et Huet is a traditional Uygur herbal medicine that has been used as a cholagogic and diuretic agent to improve liver function. However, the mechanism is not known for the liver-protective function. We investigated the antioxidant effects of plant extraction (CGE60) in vitro and in vivo, and find the mechanism of liver protection in Bacille Calmette-Guerin vaccine (BCG)+Lipopolysaccharides (LPS) induced liver injury in mice.

Materials and methods

CGE60 was made, and the antioxidant activity was investigated by comparing the ability of scavenging 1,1-diphenyl-2-picrylhydrazyl (DPPH), and 2,2-azinobis(3-ehtylbenzothiazolin-6-sulfnicAcid) diammonium salt (ABTS) free radicals in vitro. Then, CGE60 was administrated in mice of liver damage model which was induced in mice using the BCG+LPS protocol. The CGE 60 extract was tested at three dosages: 50 mg/kg, 100 mg/kg, and 200 mg/kg. Product of lipid peroxidation (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX,), nitric oxide (NO), nitric oxide synthetase (NOS), hydroxyproline and alkaline phosphatase (ALP) contents were evaluated in liver to determine the CGE60 activity in the hepatic injury model. Tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and transforming growth factor-β (TGF-β) proteins were determined in the liver tissues using ELSIA. The signaling activities were evaluated in Western blot.

Results

CGE60 exhibited strong antioxidant ability in vitro. With oral administration, CGE60 significantly increased the activity of CAT, SOD, GSH-PX, and decreased the level of NO, NO synthase, hydroxyproline, ALP and lipid peroxidation liver of in the BDG+ LPS model. CGE60 attenuated hepatic inflammation via down- regulation of TNF-α, IL-6 and TGF-β. CGE60 blocked protein expression of cytochrome P450 2E1 (CYP2E1), nuclear factor kappa-B (NF-κB), phosphorylation of extracellular signal-regulated kinase (p-ERK1/2), and cyclooxygenase-2 (COX-2),but activated the expression of p-P38 MAPK.

Conclusion

This study suggests that CGE60 possesses antioxidant activity and this activity associates with hepatoprotective effect in the mice of BCG +LPS model, and the mechanisms underlying these effects may involve antioxidant actions and anti-inflammation activities.     

Suppressive effects of wild bitter gourd (Momordica charantia Linn. var. abbreviata ser.) fruit extracts on inflammatory responses in RAW 264.7 macrophages

Aim of the study

Bitter gourd (Momordica charantia) is used to treat various diseases including inflammation. A wild species of bitter gourd, Momordica charantia Linn. var. abbreviata ser. (WBG), is considered to be more potent in disease prevention than is bitter gourd; however, little is known about the biological and physiological characteristics of WBG.

Materials and methods

The present study investigated the anti-inflammatory effect of WBG on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages.

Results

Among the hot water, 95% ethanol, and ethyl acetate extracts of WBG, the ethanol extract showed the greatest reduction of LPS-induced nitric oxide (NO) and prostaglandin E₂ (PGE₂) production and inducible nitric oxide synthase (iNOS) and pro-interleukin-1β expression. LPS-induced cyclooxygenase-2 expression was not affected by WBG extracts. Compared with WBG, extracts from bitter gourd showed a lesser inhibition of LPS-induced events. Electrophoretic mobility shift assay further showed that both the hot water and the ethanol extracts of WBG inhibited NF-κB activation. Although information is lacking on the bioactive components of WBG, the phenolic compound contents of each extract significantly paralleled its anti-inflammatory ability (r = 0.74, 0.88 and 0.65 for NO, PGE₂ and iNOS expression, respectively, P < 0.05).

Conclusions

These results suggest that WBG is beneficial for reducing LPS-induced inflammatory responses by modulating NF-κB activation.     

Analgesic and antimalarial activities of crude leaf extract and fractions of Acalypha wilkensiana

Aim of the study

Antiplasmodial and analgesic activities of leaf extract and fractions of Acalypha wilkensiana were evaluated to ascertain the folkloric claim of its antimalarial and analgesic activities.

Materials and methods

The crude leaf extract (220–659 mg/kg) and fractions (chloroform and aqueous; 440 mg/kg) of Acalypha wilkensiana were investigated for antiplasmodial activity against chloroquine sensitive Plasmodium berghei infections in mice and for analgesic activity against chemical and heat-induced pains. The antiplasmodial activity during early and established infections as well as prophylactic activity were investigated. Chloroquine (5 mg/kg) and pyrimethamine (1.2 mg/kg) were used as positive controls. Thin films made from tail blood of each mouse were used to assess the level of parasitaemia of the mice. Analgesic activity of the crude extract was also evaluated against acetic acid, formalin and heat-induced pains.

Results

The extract and its fractions dose-dependently reduced parasitaemia induced by chloroquine sensitive Plasmodium berghei infection in prophylactic, suppressive and curative models in mice. These reductions were statistically significant (p < 0.001). They also improved the mean survival time (MST) from 16 to 22 days relative to control (p < 0.01–0.001). The activities of extract/fractions were incomparable to that of the standard drugs used (chloroquine and pyrimethamine). On chemically and thermally induced pains, the extract inhibited acetic acid and formalin-induced inflammation as well as hot plate-induced pain in mice. These inhibitions were statistically significant (p < 0.001) and in a dose-dependent fashion.

Conclusion

The antiparasitaemic and analgesic effects may in part be mediated through the chemical constituents of the plant.     

Antiinflammatory and antinociceptive activities of Blechnum occidentale L. extract

Aim of study

Blechnum occidentale L. is a terrestrial fern that ranges from the United States to South America, and is employed in Brazilian folk medicine. In the present study we investigated the antinociceptive and antiinflammatory activities of the methanolic extract of Blechnum occidentale L. (MEB) in animal models of pain and inflammation to support its medicinal use in treatment of inflammatory and pulmonary diseases, urinary infections and liver diseases.

Materials and methods

The antinociceptive activity of MEB was evaluated using the writhing, formalin, and tail flick tests. The antiinflammatory activity of MEB was evaluated in carrageenan-induced paw oedema and neutrophil migration. In order to discard possible non-specific muscle relaxant or sedative effects of MEB, mice motor performance was evaluated in the rota rod test and its toxicity evaluated over 14 days.

Results

Intraperitoneal (IP) administration of MEB (0.01–100 mg/kg) produced a dose-related antinociception on acetic acid-induced writhing in mice. Oral administration of MEB, at a different range of doses (100–400 mg/kg), also produced significant antinociceptive effect on the writhing test. Furthermore, treatment with MEB (100 and 200 mg/kg IP) inhibited significantly both the early and late phases of formalin-induced hypernociception in rats. In contrast, treatment with MEB (100 and 200 mg/kg IP) did not prevent the thermal nociception in the tail flick test. The IP administration of MEB (100 and 300 mg/kg) significantly reduced the paw oedema induced by carrageenan. Moreover, systemic treatment with MEB (11–300 mg/kg) reduced the neutrophil migration in the carrageenan-induced migration to the peritoneal cavity. In the rota rod test, MEB-treated mice did not show any significant motor performance alterations with the dose of 300 mg/kg. In addition, over the study duration of 14 days, there were no deaths or toxic signs recorded in the mice given 100 or 1000 mg/kg of MEB.

Conclusion

The results described here are the first report of pharmacological studies of Blechnum occidentale L. and indicate that this plant has antinociceptive and antiinflammatory activities which support its folk medicine use.