Differential role of YiGanKang decoction in IL-1β induction of IL-1RI and AP-1 in rat hepatic stellate cell

Background/Aim

YiGanKang, a combination of Chinese herbs, has anti-fibrosis effects in chronic liver diseases. The present study tries to demonstrate differential role of YiGanKang Decoction in interleukin-1β (IL-1β) induction of the type I receptor (IL-1RI) and the activator protein 1 (AP-1) in rat hepatic stellate cell (HSC).

Methods

Flow cytometry was used to detect the IL-1RI expression. Electrophoretic mobility shift assays was used to detect AP-1 activity.

Results

IL-1RI expression after IL-1β treatment for 0, 2, 10, 60, and 120 min was 227.08 (13.15), 268.43 (8.93), 442.97 (7.00), 367.66 (14.70), and 261.58 (15.08), respectively. The differences between IL-1RI expression after treatment for 10 and 60 min were significantly higher than the corresponding values in the control (P < 0.01, P < 0.01, respectively); After pretreatment with YiGanKang Decoction, IL-1RI expression induced by IL-1β was not decrease obviously; IL-1β could activate AP-1 in rat HSCs (P < 0.01). Meanwhile YiGanKang Decoction could inhibit activity of AP-1 induced by IL-1β (P < 0.01), and the inhibition rate was 42.71%.

Conclusion

YiGanKang Decoction could not decrease IL-1RI expression, but it could inhibit activity of AP-1 in rat HSCs induced by IL-1β.     

Inhibitory effects of the root extract of Dipsacus asperoides C.Y. Cheng et al T.M.Ai on collagen-induced arthritis in mice

Ethnopharmacological relevance

Dipsaci radix, the dried root of Dipsacus asperoides C.Y. Cheng et al T.M.Ai is used as a medicinal plant in oriental clinics for the treatment of bone diseases and functions by strengthening bone and healing bone fractures.

Aim of the study

This study investigated the therapeutic efficacy of Dipsaci radix in treating rheumatoid arthritis using a type II collagen (CII)-induced arthritis (CIA) mouse model.

Materials and methods

Arthritis was induced in male DBA/1 mice by immunization with CII. Dipsaci radix water (DR-W) extract at 50 mg/kg and 100 mg/kg was orally administered from days to after the induction of arthritis. Arthritic score, serum levels of anti-CII IgG2a, the inflammatory mediator prostaglandin E₂ (PGE₂), and inflammatory cytokines (TNF-α, IL-1β and IL-6), and histological changes in the ankle joint were analyzed in CIA mice.

Results

Arthritic induction increased the arthritic score, as well as serum levels of anti-CII IgG2a antibody, PGE₂, TNF-α, IL-1β and IL-6 in mice. However, administration of DR-W extract in CIA mice significantly reduced arthritic scores and serum levels of anti-CII IgG2a antibody, PGE₂, TNF-α, IL-1β and IL-6 compared with those in vehicle-treated CIA mice. Furthermore, histopathological improvement in joint architecture was also observed in DR-W extract-treated CIA mice.

Conclusions

DR-W extract has anti-inflammatory and anti-arthritic effects in arthritic mice. This suggests that Dipsaci radix might be used as a therapeutic agent for the treatment of human arthritis.     

Effect of Salvianolic-acid B on inhibiting MAPK signaling induced by transforming growth factor-β1 in activated rat hepatic stellate cells

Aim of the study

Salvianolic-acid B (SA-B) is an effective component of Radix Salviae miltiorrhizae for anti-hepatic fibrotic herbs. MAPK signaling pathway has been implicated in hepatic stellate cells (HSC) stimulated by TGF-(1. We have investigated the effect of SA-B on MAPK pathway in rat HSC.

Materials and methods

To observe the pharmacological effect of SA-B on HSC, SA-B was added into the medium of primary HSC. TGF-(1 was added during last 2 h, and PD98059 (ERK inhibitor) and SB203580 (p38 inhibitor) were added just 30 min before adding TGF-(1. MEF2 and Col. I were measured by luciferase reporter gene assay and Western blot. (-SMA, MEF2, Raf, ERK, p-ERK, MEK, p-MEK, p38, p-p38, MKK3 and p-MKK3/6 were assayed by Western blot. Activity of MMP-2 and MMP-9 was analyzed by zymography. Each experiment was repeated for three times.

Results

The expression of (-SMA and Col. I in HSC was inhibited by SA-B. There was no effect of SA-B on the activity of MMP-2 or MMP-9 in the media of cultured HSC. Phosphorylation of ERK1/2 in HSC stimulated with or without TGF-(1 was inhibited by SA-B. Specifically, phosphorylation of MEK (upstream kinase of ERK pathway) was inhibited by SA-B. SA-B also inhibited phosphorylation of MKK3/6 (upstream kinases of p38 pathway) and inhibited the synthesis of MEF2.

Conclusions

SA-B performs anti-hepatic fibrosis through inhibiting ERK and p38 MAPK pathway in HSC. SA-B inhibits ERK pathway via inhibiting phosphorylation of MEK and inhibits p38 MAPK pathway via blocking phosphorylation of MKK3/6 and inhibiting expression of MEF2 in HSC with or without TGF-(1 stimulation.     

Sophocarpine administration preserves myocardial function from ischemia-reperfusion in rats via NF-κB inactivation

Ethnopharmacological relevance

Sophora alopecuroides L. (the clinical usefulness of compound Kudouzi injection) has been used mainly for the treatment of fever, inflammation, edema and pain. Sophocarpine, a tetracyclic quinolizidine alkaloid, is one of the most abundant active ingredients in Sophora alopecuroides L. Sophocarpine injection (called the Kangke injection) has been demonstrated to have significant antivirus effects against coxsackievirus B3 and therapeutic effects for viral myocarditis in clinical.

Aim of the study

The present study was to evaluate the protective effect of sophocarpine on the inhibition of NF-kappaB (NF-κB) and effect on inflammatory markers during myocardial ischemia-reperfusion (I/R) injury in rat.

Materials and methods

Myocardial I/R injury was induced by the occlusion of left anterior descending coronary artery for 30 min followed by reperfusion for 2 h. 2 h after reperfusion was established, the hemodynamics and infarct size were examined. Blood samples were collected for biochemical analysis. Expression of NF-κB and mitogen-activated protein kinases (MAPKs) in ischemic myocardial tissue were assayed by western blot.

Results

Administration of sophocarpine significantly improved cardiac function and reduced infarct size in I/R rat heart in vivo. Furthermore, sophocarpine ameliorated the contents of inflammatory mediators (tumor necrosis factor-alpha, TNF-α; interleukin-6, IL-6; IL-10), neutrophil infiltration and myeloperoxidase (MPO) activity. Interestingly, sophocarpine also significantly inhibited translocation of NF-κB, which was associated with attenuated phosphorylations of p38 and c-Jun NH2-terminal protein kinase (JNK).

Conclusions

Inflammatory mediators, infiltration of neutrophil, and MPO were ameliorated via down-regulation of JNK and p38, and inactivation of NF-κB. This might be one of the important mechanisms of sophocarpine that protected myocardial injury from I/R.     

Euonymus alatus extract attenuates LPS-induced NF-κB activation via IKKβ inhibition in RAW 264.7 cells

Aim of the study

The present study was performed to investigate the underlying mechanisms of anti-inflammatory effects with the extract of Euonymus alatus (EEA), and specially focused on nuclear factor κB (NF-κB) signaling pathway by targeting the IκB kinase β (IKKβ).

Materials and methods

The effect of EEA for IKKβ activity was analyzed using an immobilized metal affinity for phosphochemicals (IMAP)-based time-resolved fluorescence resonance energy transfer (TR-FRET) assay. The effect of EEA on lipopolysaccharide (LPS)-induced NF-κB activation in murine macrophage RAW 264.7 cells with western blotting and immunofluorescent staining was evaluated.

Results

IKKβ studies based on IMAP-TR-FRET showed that EEA possesses a potent IKKβ inhibitory activity with IC₅₀ value of 11.83 μg/ml. EEA (10, 30 μg/ml) also attenuated the LPS-induced IκBα phosphorylation/degradation, NF-κB translocation and subsequent NO synthesis in RAW 264.7 cells.

Conclusions

These results suggest that EEA abrogates LPS-induced NF-κB signaling pathway by targeting the IKKβ in RAW 264.7 cells and these properties may provide a molecular basis for understanding the inhibitory effects of EEA on LPS-mediated inflammation.     

Icaritin induces cell death in activated hepatic stellate cells through mitochondrial activated apoptosis and ameliorates the development of liver fibrosis in rats

Aim of the study

Icaritin is an active ingredient extracted from the plant Herba Epimedium Sagittatum (Sieb. et Zucc.) Maxim. The purpose of this study is to investigate the effects and mechanisms of icaritin-induced cell death in activated hepatic stellate cells (HSCs) and ameliorating the development of liver fibrosis in rats.

Materials and methods

: In vitro, icaritin-induced cell death rates in HSC-T6 (rat) and LX-2 (human) HSC lines as well as normal hepatocyte cell lines HL-7702 (L02) and WRL-68 were assayed by MTT method, and the apoptotic ratios were detected by both flow cytometry and the Annexin-V-FITC Apoptosis Detection Kit. A Whole Rat Genome Microarray Kit was used to identify expression of interest genes through fold-change screening. In vivo study, experimental liver fibrosis models were built by carbon tetrachloride (CCl₄) or common bile duct ligation (CBDL) in Wistar rats. Icaritin (1 mg/kg/day, three days a week) was administered by gastric gavage for four weeks (n = 6 per group). At the end of the study, serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) as well as the contents of hydroxyproline and collagen I in liver tissues were measured. Histopathological changes and the distribution of activated HSCs were observed in the liver tissues using hematoxyline-eosin (HE) staining and immunohistochemical staining for α-smooth muscle actin (α-SMA).

Results

Icaritin induced apoptosis in HSC-T6 and LX-2 in a concentration- and time-dependent manner with little toxicity to normal hepatocyte cell lines. The IC₅₀ of icaritin in HSC-T6 was 12.83 μM at 48 h. Apoptotic ratio of HSC-T6 treated with 24 μM icaritin was 20.19%, and the G2 phase of the cell cycle did not occur (P < 0.05). Gene analysis showed that icaritin up-regulated Bak-1, Bmf and Bax expression while significantly down-regulated Bcl-2 expression (vs. control group, P < 0.01). These results suggested that mitochondrial pathway played an important role in icaritin-induced apoptosis in activated HSCs. In vivo results showed that icaritin reduced the number of activated HSCs, and brought the elevated levels of AST, ALT, hydroxyproline and collagen I to normal or near normal values (vs. model group, P < 0.05).

Conclusions

Icaritin can induce cell death in activated HSCs through mitochondria-mediated apoptosis, ameliorate the progression of hepatic fibrosis in rats, and could be a promising drug for treating liver fibrosis.     

Suppression of interleukin (IL)-8 and human beta defensin-2 secretion in LPS-and/or IL-1β-stimulated airway epithelial A549 cells by a herbal formulation against respiratory infections (BNO 1030)

Aim of the study

A special ethanolic-aqueous extract from seven traditional medicinal plants (BNO 1030) has been used for several decades to treat recurrent infections of the respiratory tract. Considering the potential role of interleukin-8 (IL-8) and human beta defensin-2 (hBD-2) in inflammation, we investigated the effect of BNO 1030 on lipopolysaccharide (LPS) from Pseudomonas aeruginosa or IL-1β-induced inflammatory mediators in A549 human type II alveolar epithelial cells.

Materials and methods

A549 cells were stimulated with LPS (100 μg/ml) or IL-1β (50 ng/ml) in the presence of the preparation and the secretion of IL-8 and hBD-2 were measured after 18 h and 24 h in cell free supernatants using enzyme-linked immunosorbent assays (ELISA). Cell viability and cell growth was investigated by propidium iodide uptake and WST-1 assay, respectively.

Results

BNO 1030 inhibited the secretion of IL-8 and hBD-2 at non-cytotoxic concentrations (0.1-100 μg/ml; cell growth inhibitory concentration, 50% (IC₅₀) = 678 ± 87.6 μg/ml). Stimulation by IL-1β led to a 7-fold activation of IL-8 secretion, which was reduced by 37.7 ± 4.1% (p < 0.05) after incubation with 100 μg/ml BNO 1030. Inducible hBD-2 was suppressed by 91.8 ± 15.6% (p < 0.01) at the same concentration of BNO 1030 (IC₅₀ = 0.7 ± 0.1 μg/ml). The 2-fold increase of IL-8 secretion by LPS-stimulated cells was completely abolished at concentration of 50 μg/ml BNO 1030 (IC₅₀ = 5.7 ± 3.6 μg/ml).

Conclusion

BNO 1030 suppressed the secretion of IL-8 and hBD-2 in cultured epithelial A549 cells. These results support its use as a phytotherapeutic product prepared from traditional remedies in inflammatory diseases, especially those affecting the respiratory tract.     

Kirenol upregulates nuclear annexin-1 which interacts with NF-κB to attenuate synovial inflammation of collagen-induced arthritis in rats

Ethnopharmacological relevance

Kirenol is a diterpenoid compound purified from the Chinese Herba Siegesbeckiae. Siegesbeckiae has been employed for the treatment of arthritis for centuries, its safety and efficacy are documented through a long history of human use.

Aim of the study

To investigate the effects on collagen-induced arthritis (CIA) and anti-inflammatory mechanism of Kirenol.

Materials and methods

Kirenol was administrated intragastrically in rats after the onset of CIA. Pathological changes were evaluated by paw swelling and histopathology. Concentration of IL-1β in synovial fluid and adrenal corticotropin (ACTH) in plasma were determined by Elisa. Western blot was performed to detect the expression of Annexin-1 and glucocorticoid receptor alpha (GRα) in synovium. NF-κB DNA binding activity was assessed by electrophoretic mobility shift assays (EMSA).

Results

Kirenol (1, 2, and 4 mg/kg) and Prednisolone depressed paw swelling and reduced IL-1β of synovial fluid in the CIA rats (p < 0.05 or p < 0.01). Kirenol and Prednisolone upregulated nuclear Annexin-1 and inhibited NF-κB activity in synovium of CIA. The inhibitory effect of Kirenol and Prednisolone on NF-κB activity was enhanced by anti-Annexin-1 Ab. Prednisolone, but not Kirenol, downregulated plasma ACTH and GRα expression significantly (p < 0.01).

Conclusion

Kirenol and Prednisolone can upregulate nuclear Annexin-1 which interacts with NF-κB to inhibit NF-κB activity, reduce cytokines expression and thereby attenuate inflammation of CIA joints. Kirenol does not lead to ACTH or GR downregulation, which is in contrast to classic glucocorticoid Prednisolone. Kirenol shares with GCs similar anti-inflammatory mechanism but bypass the considerable limitation of GCs treatment.     

Effects on neuroendocrinoimmune network of Lizhong Pill in the reserpine induced rats with spleen deficiency in traditional Chinese medicine

Aim of the study

Lizhong Pill, composed of Radix Ginseng (Panax ginseng C.A. Meyer), Rhizoma Zingiberis (Zingiber officinale Roscoe), Rhizoma Atractylodis Macrocephalae (Atractylodes macrocephala Koidz.) and Radix Glycytthizae (Glycyrrhiza uralensis Fisch.), is a classical herbal product for curing spleen deficiency in traditional Chinese medicine (TCM), and reserpine treated rats show similar signs to TCM spleen deficiency pattern. This paper is aimed to explore the regulatory effect on neuroendocrinoimmune network by Lizhong Pill in reserpine induced TCM spleen deficiency rats.

Materials and methods

100 healthy adult male SD rats, with a mean weight of 200 g, were randomly divided into five groups in average: control group, reserpine treated group, atropine treated group, treatment groups with Lizhong Pill at high dose and low dose (equal to the dosage of crude drugs for 4 g/kg/d and 8 g/kg/d). Rats in reserpine treated group were induced by intraperitoneal injection of reserpine at 0.5 mg/kg d for 4 weeks. The levels of IL-1, IL-6 and gastrin were measured with radioimmunoassay, TNF-α and IFN-γ in serum were measured with ELISA, the level of vasoactive intestinal peptide (VIP) and substance P (SP) in small intestine were determined with radioimmunoassay, and the TNF-α and TGF-β positive cells in small intestine were detected by immunohistological staining. Data were analyzed with SAS 9.1 software package.

Results

The rats in reserpine treated group, body weight, concentrations of IFN-γ, IL-1 and TNF-α in serum, expression of TGF-β in small intestine, VIP in small intestine decreased (P < 0.05), and the level of IL-6 in serum, expression of TNF-α, SP in small intestine and gastrin were increased (P < 0.05). Administration of Lizhong Pill at high dose could increase the body weights at day 21, and the weights of rats in Lizhong Pill groups were much higher compared to reserpine treated group. At high dose of Lizhong Pill could increase the level of TNF-α in serum. Lizhong Pill at high dose and low dose could reverse the changes of IL-1, IL-6 and IFN-γ, gastrin, expression of TGF-β and TNF-α, VIP and SP in small intestine.

Conclusions

The rats treated with reserpine, with similar signs to TCM spleen deficiency, show neuroendocrinoimmune disorders, and the restoration of the neuroendocrinoimmune disorders might be the part of mechanism of Lizhong Pill for reinforcing TCM spleen deficiency.     

Modified Si-Miao-San extract inhibits inflammatory response and modulates insulin sensitivity in hepatocytes through an IKKβ/IRS-1/Akt-dependent pathway

Aim of the study

Modified Si-Miao-San (mSMS) has showed anti-inflammatory potency and has been used in the clinic to treat metabolic disorders such as obesity and diabetes, but whether its anti-inflammatory activity contributes to improving insulin resistance remains to be determined. This study aims to investigate the mechanistic relationship between its anti-inflammatory activity and modulation of insulin sensitivity in free fatty acid-stimulated HepG2 cells.

Materials and methods

HepG2 cells were stimulated with palmitate (PA) and the effect of mSMS on insulin mediated-glycogen synthesis and triglyceride secretion was observed. The inhibition of mSMS on gene expression of proinflammatory cytokine tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and inhibitor of NF-κB kinase-β (IKKβ) activation was investigated. In addition, the effects of mSMS on insulin signaling transduction along insulin receptor substrates-1 (IRS-1)/Akt pathway were also evaluated. Furthermore, the effect of mSMS on glucose intolerance induced by conditioned-medium derived from activated macrophages was also assessed in normoglycemic mice.

Results

Treatment of hepatocytes with PA reduced insulin sensitivity and mSMS effectively increased insulin-mediated glycogen synthesis and restored insulin inhibition of triglyceride secretion. mSMS suppressed IKKβ activation and down-regulated TNF-α and IL-6 gene over-expression, demonstrating its anti-inflammatory activity in hepatocytes. PA-evoked inflammation impaired insulin signaling cascades and mSMS improved insulin signaling transduction by modification of Ser/Thr phosphorylation of IRS-1 and downstream Akt (T308), thereby improved insulin sensitivity in hepatocytes. mSMS also improved glucose intolerance induced by inflammatory cytokines in normoglycemic mice, which further demonstrated its modulation toward insulin sensitivity in vivo.

Conclusions

The results suggest that mSMS inhibited inflammatory response and improved insulin sensitivity in hepatocytes via an IKKβ/IRS-1/Akt-dependent pathway.     

Sheng-Ma-Ge-Gen-Tang (Shoma-kakkon-to) inhibited cytopathic effect of human respiratory syncytial virus in cell lines of human respiratory tract

Ethnopharmacological relevance

Sheng-Ma-Ge-Gen-Tang (SMGGT; Shoma-kakkon-to) has been used against pediatric viral infection for thousands of year in ancient China. However, it is unknown whether SMGGT is effective against human respiratory syncytial virus (HRSV).

Aim of the study

HRSV is a major pediatric viral pathogen of low respiratory tract infection without effective management. This study tested the hypothesis that SMGGT effectively inhibited cytopathy induced by HRSV.

Materials and methods

Effect of the crude extract of SMGGT on HRSV was tested by plaque reduction assay in both human upper (HEp-2) and low (A549) respiratory tract cell lines. Ability of SMGGT to stimulate anti-viral cytokines was evaluated by enzyme-linked immunosorbent assay (ELISA).

Results

Crude extract of SMGGT dose-dependently inhibited HRSV-induced plaque formation. The crude extract was more effective when given before viral infection (p < 0.0001). It inhibited viral attachment dose-dependently (p < 0.0001) and could increase heparin effect on viral attachment. Furthermore, it was synergistic with very low-dose heparin on viral attachment. In addition, the crude extract time-dependently and dose-dependently (p < 0.0001) inhibited HRSV internalization into HEp-2 cells. Epithelial cells secrete IFN-β and TNF-α to counteract viral infection. The crude extract could stimulate epithelial cells to secrete these cytokines beforehand and become resistant to viral infection. It also stimulated IFN-β to defense HRSV after viral inoculation.

Conclusions

Sheng-Ma-Ge-Gen-Tang could be effective to manage HRSV infection in young children.     

Effects of Scutellariae radix and Aloe vera gel extracts on immunoglobulin E and cytokine levels in atopic dermatitis NC/Nga mice

Aim of the study

The present study aimed to investigate the effects of Scutellariae radix (SR) and Aloe vera gel (AV), alone or in combination, on levels of immunoglobulin E (IgE) and inflammatory cytokines in spontaneous atopic dermatitis(AD)-like skin lesions.

Materials and methods

After spontaneous AD-like skin lesion was developed by adaptation to conventional conditions, mice were randomly assigned to control, SR (50 mg/kg, p.o.), AV (0.8 mg/kg, p.o.) and SRAV (50 mg of SR and 0.8 mg of AV/kg, p.o.) groups, and were treated for 6 weeks.

Results

SR and SRAV suppressed IL-5 levels compared with control, but had no effects on IgE levels (P < 0.05). AV increased IgE levels, but decreased both IL-5 and IL-10 compared with control (P < 0.05).

Conclusion

These results suggest that SR and AV modulate immunological responses in AD, mainly through influencing IL-5 or IL-10 levels.     

Biphasic effects of baicalin, an active constituent of Scutellaria baicalensis Georgi, in the spontaneous sleep-wake regulation

Aim of the study

Baicalin is an active compound originating from the root of Scutellaria baicalensis Georgi, which has been used for anti-inflammation, anti-bacteria, anti-hypertension, anti-allergy and sedation since ancient China, though the neuronal mechanisms involved in the sedative effect is still unclear. Baicalin possesses the ability to decrease the expression of pro-inflammatory cytokines and nuclear factor (NF)-κB activity. Furthermore, baicalin has demonstrated an anxiolytic-like effect via activation of γ-aminobutyric acid-A (GABA_A) receptors. Pro-inflammatory cytokines (e.g. interleukin (IL)-1, IL-6, and tumor necrosis factor (TNF)-α) and the GABAergic system promote sleep. This study was designed to determine whether the GABA_A receptor activation and/or the suppression of pro-inflammatory cytokines mediate(s) baicalin-induced sleep alterations.

Materials and methods

Baicalin was intracerebroventricularly (ICV) administered 20 min either prior to the beginning of the light period or before the onset of the dark period. Electroencephalogram (EEG) and gross body movement were acquired for sleep analysis. Pharmacological blockade of IL-1 and GABA_A receptors were employed to elucidate the involvements of IL-1 and GABA_A receptors in baicalin-induced sleep alterations. IL-1β concentrations obtained after baicalin administration in several distinct brain regions were determined by ELISA.

Results

ICV administration of baicalin decreased slow wave sleep (SWS) during the first 2 h of the light period. Rapid eye movement sleep (REMS) was not altered. The blockade of IL-1β-induced SWS enhancement by baicalin suggests that the antagonism of IL-1 receptors is involved in baicalin-induced SWS decrement during the light period. However, IL-1β concentrations during the light period were not altered after baicalin administration. In contrast, baicalin increased both SWS and REMS during hours 8-10 of the dark (active) period when baicalin was administered at the beginning of the dark period, and its effects were blocked by the GABA_A receptor antagonist bicuculline.

Conclusion

Baicalin exhibits biphasic effects on sleep-wake regulation; the decrease of SWS during the light period and increases of SWS and REMS during the dark period. Inhibition of IL-1 action and enhancement of GABA_A receptor activity may mediate baicalin's effects during the light and dark period, respectively.     

Protective effect of a polyphenolic rich extract from Magnolia officinalis bark on influenza virus-induced pneumonia in mice

Ethnopharmacological relevance

Magnolia officinalis bark is used in traditional Chinese medicine for the treatment of cough, colds, fever, chronic bronchitis and stomach ailments.

Aim of the study

To investigate therapeutic effects of polyphenol rich extract from M. officinalis bark (MPE) in influenza virus A-infected mice, and to provide evidence for the inflammation response and immunomodulatory potential during infection.

Materials and methods

Mice were infected with influenza virus A (IVA) and MPE at doses of 10 and 20 mg/kg were orally administrated daily for 5 days after challenge. The levels of serum L-6 and TNF-α were determined by ELISA while protein expressions of NF-κB and TLR3 were detected by western blotting analysis.

Results

MPE exhibited significant therapeutical effects on reducing levels of serum NO, IL-6 and TNF-α, inhibiting pneumonia, decreasing lung viral titers and sensitizing IVA-induced apoptosis through down-regulation of NF-κB and TLR3 protein expression in the lung tissue of IVA-infected mice.

Conclusions

MPE could exhibit preventive and therapeutical effects on IVA-infected mice as a suppressor of the production of inflammatory mediators, NO and pro-inflammatory cytokines, TNF-α and IL-6. These effects appeared to be mediated, at least in part, by an inhibition of TLR3 and NF-κB activation. Therefore, MPE could provide a safe and effective therapeutic approach for influenza and its subsequent viral pneumonia.     

Antinociceptive and gastroprotective actions of ethanolic extract from Pluchea sagittalis (Lam.) Cabrera

Ethnopharmacological relevance

Pluchea sagittalis, an herbaceous plant widely distributed in South America, is used in folk medicine for the treatment of digestive diseases and inflammation.

Aim of the study

This study was designed to investigate the antinociceptive and gastroprotective effects of the ethanolic extract (EE) of aerial parts from Pluchea sagittalis in rodents.

Materials and methods

The antinociceptive effects of EE was evaluated in mice after oral administration in chemical tests (acetic-acid, glutamate and formalin) or by biting behavior following intrathecal administration of cytokines such as interleukin-1beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) in mice. Furthermore, rats were treated with EE and subsequently exposed to acute gastric lesions induced by 80% ethanol. Afterwards the gastric lesion extension and the mucus levels of gastric mucosa were measured.

Results

The oral administration of EE showed a dose-dependent inhibition of acetic acid-induced abdominal constrictions and glutamate-induced pain in mice, with ID₅₀ values of 624.0 (523.0-746.0) mg/kg and 368.0 (216.0-628.0) mg/kg, respectively. In the formalin test, the EE also produced significant inhibition of the inflammatory phase, with an ID₅₀ value of 411.0 (183.0-721.0) mg/kg; however, it was ineffective in the neurogenic phase caused by formalin. In addition, oral treatment with EE caused a significant inhibition of biting behavior induced by i.t. injection of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α). The antinociception caused by the EE (300 mg/kg, p.o.) was not reversed by naloxone (1 mg/kg, i.p.) when assessed in the acetic acid writhing test. The EE (300-1000 mg/kg, p.o.) did not affect the motor coordination of animals in an open-field model. Oral treatment with the EE protected rats against gastric lesions induced by ethanol, with an ID₅₀ value of 55.0 (46.6-64.9) mg/kg, and increased the mucus levels of gastric mucosa to levels found in the non-lesioned group.

Conclusions

The mechanism by which the extract produced antinociception still remains unclear, but this effect seems to be primarily related to the modulation or inhibition of the action of pro-inflammatory mediators. Furthermore, these data support, at least in part, the ethnomedical use of Pluchea sagittalis.     

Antinociceptive and antiinflammatory activities of Adiantum latifolium Lam.: Evidence for a role of IL-1β inhibition

Aim of study

Adiantum, one of the most widely distributed genera of the family Pteridaceae, is employed in folk medicine worldwide. Adiantum latifolium Lam. has been used in Latin American traditional medicine as anxiolytic, analgesic and antiinflammatory. The present study investigates the antinociceptive and antiinflammatory properties of the methanolic extract of Adiantum latifolium (MEA) in animal models of pain and inflammation to confirm its medicinal use.

Material and methods

The antinociceptive and antiinflammatory activities of MEA were evaluated using the writhing, formalin, and tail-flick tests, carrageenan-induced paw edema and arachidonic acid-induced ear edema. Mice motor performance was evaluated in the rota rod test and the acute toxicity evaluated over 14 days.

Results

Intraperitoneal (1-100 mg/kg) or oral (100-400 mg/kg) administration of MEA produced a dose-related inhibition of acetic acid-induced writhing in mouse. Furthermore, treatment with MEA (100 mg/kg) inhibited both the early and late phases of formalin-induced hypernociception. In contrast, MEA (100 mg/kg/IP) did not prevent the thermal nociception in the tail-flick test. In addition, MEA (100 and 200 mg/kg/IP) inhibited important events related to the inflammatory response induced by carrageenan or arachidonic acid, namely local edema and increase in tissue interleukin-1β levels. MEA (300 mg/kg/IP)-treated mice did not show any motor performance alterations. Over the study period of 14 days, there were no deaths or toxic signs recorded in the group of mice given 1000 mg/kg of MEA.

Conclusion

The results demonstrate that Adiantum latifolium has antinociceptive and antiinflammatory activities, acting through the inhibition of IL-1β production.     

Anticonvulsant activity of the methanolic extract of Justicia extensa T. Anders

Ethnopharmacological relevance

To investigate the anticonvulsant activity of the leaf extract of Justicia extensa T. Anders used traditionally in the treatment of convulsion.

Materials and methods

The anticonvulsant activity of the methanolic extract of Justicia extensa (50, 100 and 200 mg/kg, p.o.) was assessed in strychnine-induced (STR) and picrotoxin-induced (PCT) convulsion models in mice. Diazepam (1 mg/kg) and phenobarbitone (2 mg/kg) were used as reference drugs respectively.

Results

The extract showed no toxicity and significantly prolonged (p < 0.01-0.05) the onset and reduced the duration of the seizures induced by picrotoxin (5 mg/kg, i.p.) in a dose dependent manner. Phenobarbitone completely inhibited the seizures in this model. Similarly, in the seizures induced by strychnine (1 mg/kg, i.p.), the extract also prolonged the onset and reduced the duration of the seizures though not in a dose dependent manner. Diazepam failed to inhibit the strychnine-induced seizures. The plant extract however showed a significantly higher anticonvulsant activity at 100 and 200 mg/kg in comparison with diazepam.

Conclusions

The results obtained from this work suggest that Justicia extensa has anticonvulsant activity and this supports the use of the plant traditionally in the treatment of convulsion.     

Hepatoprotective potential of Tecomella undulata stem bark is partially due to the presence of betulinic acid

Ethnopharmacological relevance

As a well-known Chinese Materia Medica Rhizoma Anemarrhenae has multiple pharmacological activities including antipyretic, anti-inflammatory, anti-diabetic actions, etc. This study was designed to investigate effects of total saponins from Rhizoma Anemarrhenae (TS) on diabetes-associated cognitive decline in rats and influence on amyloid-beta (Aβ) levels in brain and inflammation.

Materials and methods

Diabetic rats induced by intraperitoneal administration of streptozotocin, were randomized into two groups: diabetes and TS-treated diabetes. Blood glucose and body weight were measured monthly and weekly, respectively. After seven weeks, cognitive performances were evaluated with Morris water maze. Then, brain was obtained for assay of Aβ and TNF-α levels, and blood was collected for TNF-α assay.

Results

Aβ(1-40), Aβ(1-42) and TNF-α levels were dramatically (all P < 0.01) increased both in temporal cortex and hippocampus of diabetic rats, coupled with impairment of cognition, compared with those of the control. Chronic TS (200 mg/kg) treatment markedly (P < 0.05) improved the learning ability of diabetic rats, and significantly (all P < 0.05) reduced Aβ(1-40), Aβ(1-42) and TNF-α levels in cortex as well as Aβ(1-40) level in hippocampus, whereas showed a decreased tendency for Aβ(1-42) and TNF-α levels in hippocampus. Moreover, eight-week treatment with TS remarkably (P < 0.05) inhibited the elevation of TNF-α level in serum of diabetic rats, and significantly (both P < 0.01) decrease the fasting blood glucose level and increase the body weight of diabectic rats.

Conclusion

Our findings demonstrate that diabetes-associated cognitive decline is, at least in part, due to brain Aβ accumulation in diabetic condition, and efficacy of TS to diabetes-associated cognitive decline in rats is a sum of reduction of Aβ accumulation and inflammation in brain as well as attenuation of major symptoms of diabetes.