Sedative, hypnotic and anticonvulsant activities of the ethanol fraction from Rhizoma Pinelliae Praeparatum

Ethnopharmacological relevance

Rhizoma Pinelliae Praeparatum is the product of raw Rhizoma Pinellia processed with alkaline solution and Licorice, which had been widely used for treatment of insomnia in traditional Chinese medicine. The present study aimed to investigate the sedative, hypnotic and anticonvulsant activities of ethanol fraction from Rhizoma Pinelliae Praeparatum (EFRP) and to determine whether these effects were related to GABAergic mechanism.

Materials and methods

The sedative, hypnotic and anticonvulsant activities of EFRP were investigated with locomotion activity, pentobarbital-induced sleeping and nikethamide (NKTM)-induced convulsion tests, respectively. Additionally, the effects of flumazenil (an antagonist of GABA_A receptor) and l-malic acid (blocker of synthetic enzyme for GABA) on the hypnotic activity of EFRP were evaluated.

Results

EFRP at dose of 12 g/kg significantly inhibited the locomotion activity of mice. EFRP showed synergic effect on pentobarbital-induced sleeping by increased numbers of mice falling asleep, reduced the sleep latency and prolonged the sleeping time. l-malic acid and flumazenil inhibited the augment effects of EFRP on pentobarbital-induced sleeping. EFRP promoted a significant protection to NKTM-induced convulsion, by prolonged the death latency and decreased mortality.

Conclusion

EFRP possessed sedative, hypnotic and anticonvulsant activities and these activities may be related to the GABAergic system.     

Orally administered mycelial culture of Phellinus linteus exhibits antitumor effects in hepatoma cell-bearing mice

Aim of the study

The aim of this study was to evaluate the anticancer effect of a mycelial culture from Phellinus linteus PL-7 (MCPL-7) and to elucidate its potential mechanism in vivo.

Materials and methods

SCID CB-17 mice received a transplant of Hep3B cells followed by daily MCPL-7 administrations for 8 weeks. Following tumor implantation, groups C-E were subcutaneously administered 50 mg/kg, 100 mg/kg, or 250 mg/kg MCPL-7 powder per day, respectively, for 8 weeks. Groups A and B received saline solution subcutaneously for 8 weeks.

Results

MCPL-7 administration induced a significant reduction in tumor size and was associated with a significant increase in T cell numbers; IL-12, IFN-γ and TNF-α secretion; NK cell activity; and phagocytic ability. Therefore, increased numbers of CD4⁺cells could have been caused by greater numbers of dendritic cells and macrophages in the spleen. Furthermore, the activation of dendritic cells and macrophages resulted in increased IL-12 secretion, which could upregulate NK cell activation. The increased secretion of IL-12, IFN-γ, and TNF-α enhanced the activity and phagocytic ability of NK cells. Thus, MCPL-7 may provide a potential therapeutic approach for both immunomodulatory and antitumor effects.     

Active lipids of Ganoderma lucidum spores-induced apoptosis in human leukemia THP-1 cells via MAPK and PI3K pathways

Ethnopharmacological relevance

Ganoderma lucidum (Lingzhi) is traditionally drug, which has been traditionally effective used in the treatment of chronic hepatopathy, hypertension, hyperglycemia and cancer.

Materials and methods

THP-1 and HL-60 apoptosis induced by active lipids of Ganoderma lucidum spores was quantified by flow cytometry using FITC-conjugated annexin V and PI; MAPK and Akt were measured by Western blot, and caspase-3, -8 and -9 activities were also detected by spectrophotometric assay.

Results

Our results showed that active lipids of Ganoderma lucidum spores decreased phosphorylation-ERK1/2 (P-ERK1/2), P-Akt and increased P-JNK1/2, but did not affect expressions of P-p38 MAPK in THP-1 cells. Moreover, treatment of THP-1 cells with active lipids of Ganoderma lucidum spores resulted in activation of caspase-3, -8 and -9. Furthermore, LY294002 (Akt inhibitor) or PD98059 (ERK1/2 inhibitor) significantly enhanced active lipids of Ganoderma lucidum spores-induced apoptosis in THP-1 cells, whereas caspase inhibitors or SP600125 (JNK inhibitor), decreased apoptosis in THP-1 cells.

Conclusion

Taken together, our study for the first time suggests that active lipids of Ganoderma lucidum spores is able to enhance apoptosis in THP-1 cells, at least in part, through inhibition of ERK1/2, Akt and activation of JNK1/2 signaling pathways. Moreover, it also triggers caspase-3, -8 and -9 activation mediated apoptotic induction.     

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.     

The protective effects of total saponins from Ornithogalum saundersiae (Liliaceae) on acute hepatic failure induced by lipopolysaccharide and d-galactosamine in mice

Ethnopharmacological relevance

This study examined the protective effects of total saponins from Ornithogalum saundersiae (Liliaceae) on d-galactosamine (d-GalN) and lipopolysaccharide (LPS) - induced fulminant hepatic failure.

Materials and methods

Total saponins of Ornithogalum saundersiae (Liliaceae) (OC) were prepared with ethyl alcohol extract from bulbs of the plant. Mice were given an intraperitoneal injection of d-GalN (700 mg/kg)/LPS (10 μg/kg). OC (100 mg/kg, 200 mg/kg and 300 mg/kg) was administered orally for 3 days continuously, and at the last day at 1 h before the d-GalN/LPS injection. Mice were sacrificed at 8 h after the d-GalN/LPS injection. The liver injury was assessed biochemically, investigating aspartate aminotransferase (AST), alanine aminotransferase (ALT), malondialdehyde (MDA), glutathione (GSH) activities, and the expressions of caspase-3 and hypoxia inducible factor-1α (HIF-1α) as well. Tumor necrosis factor (TNF-α) content was measured after d-GalN/LPS induced 1 h by ELISA assay. The survival rates after application of OC in 24 h also were observed.

Results

d-GalN/LPS increased the serum aminotransferase levels and lipid peroxidation, while decreased the reduced glutathione level. The pretreatment with OC attenuated these changes in a dose-dependent manner. Elevation of TNF-α level and activation of caspase-3, HIF-1α were observed in the d-GalN/LPS group, which was attenuated by OC. The survival rate of the OC groups was significantly higher than that of the d-GalN/LPS group.

Conclusions

Protection afforded by OC against d-GalN/LPS-induced fulminant hepatic failure is the result of reduced oxidative stress, inhibited expression of caspase-3, HIF-1α, and anti-apoptotic activity.     

The differential immunological activities of Ganoderma lucidum on human pre-cancerous uroepithelial cells

Aims of the study

Ganoderma lucidum is active to stimulate immunological effector cells, but the effects on uroepithelial cells have never been explored. The present study compared the expression of major cytokines induced by the water (GLw) and ethanol (GLe) extracts of G. lucidum.

Materials and Methods

The pre-cancerous human uroepithelial cell (HUC-PC) line was employed. A total of 15 cytokines, including major Th1/Th2 cytokines and chemokines, were measured in the complete media after 24 h incubation with GLw and GLe. Additionally, the following assays were performed: cytotoxicity, apoptosis, migration of neutrophils, and nuclear factor-kappaB (NF-κB) DNA binding activity.

Results

GLe inhibited the growth of HUC-PC cells through apoptosis. Interleukins IL-2, IL-6, and IL-8 were significantly up-regulated by GLe in dose-dependent manners, but not by GLw. However, MCP-1 level was significantly increased by GLw but was oppositely reduced by GLe. Furthermore, the elevation of cytokine expression was correlated with the enhancement of p50/p65 NF-κB activity induced by GLe. The elevated IL-8 levels in GLe-treated cells were also correlated with the migration of neutrophils.

Conclusions

GLe and GLw exhibited different immunological activities on the HUC-PC cells. In particular, the activities of GLe may favor the clearance of high risk urothelial cells, suggesting potent chemopreventive ingredients are extractable by ethanol from G. lucidum.     

Macrophages, rather than T and B cells are principal immunostimulatory target cells of Lycium barbarum L. polysaccharide LBPF4-OL

Aim of the study

Lycium barbarum L. is a renowned Yin strengthening agent in traditional Chinese medicine. Lycium barbarum L. polysaccharide-protein complex is well-known for its immunoregulatory and antitumor effects. LBPF4-OL is the glycan part of Lycium barbarum L. polysaccharide-protein complex fraction 4 (LBPF4). LBPF4-OL's active contribution in LBPF4 is still blank. In the study, we enrich the polysaccharide part of Lycium barbarum L. polysaccharide-protein complex, and investigate its immunostimulatory effects on mouse spleen cells, T cells, B cells and macrophages.

Materials and methods

Balb/C mice were used in vitro and in vivo studies. In in vitro study, lymphocyte proliferations were analyzed with ³H-TdR incorporation method. Miltenyi MicroBeads were used in the purification of lymphocytes. Activation of T and B cells was analyzed by flow cytometry. In order to obtain the peritoneal macrophages, mice were injected i.p. with 1 mL of sodium thioglycollate 3 days prior to killing. Spleen cells were stimulated with LBPF4-OL and cytokine concentrations in the supernatants were determined by multiplex bead analysis. In in vivo study, mice were injected i.p. with 1 mL of normal saline or 100 μg/mL LBPF4-OL daily for 6 days. Peritoneal macrophage functions were analyzed by enzyme-linked immunosorbent assay and flow cytometry assay.

Results

Spleen cells and lymphocyte proliferation assay indicated that LBPF4-OL markedly induced the spleen cell proliferation, but could not induce proliferation of purified T and B lymphocytes. Further research revealed that B cell proliferation took place in the presence of activated macrophages or LPS. Multiplex bead analysis showed that LBPF4-OL can obviously induce IL-6, IL-8, IL-10 and TNF-α production of the spleen cells in a concentration-dependent manner. Flow cytometric analysis showed that LBPF4-OL (i.p.) prompts CD86 and MHC-II molecules expression on macrophages. ELISA assay showed that LBPF4-OL can greatly strengthen macrophage releasing of TNF-α and IL-1β.

Conclusion

These results suggested that glycan LBPF4-OL plays an important role in the immunopharmacological activity of Lycium barbarum L. polysaccharide-protein complex, and primary mouse macrophages, rather than T and B cells, are the principal target cells of it.     

Anti-inflammatory effects of methanol extract of Antrodia cinnamomea mycelia both in vitro and in vivo

Aims of the study

Antrodia cinnamomea is a folk medicinal mushroom commonly used in Taiwan for the treatment of several types of cancers and inflammatory disorders. This study aimed to explore the folk use of Antrodia cinnamomea on pharmacological grounds to characterize the scientific basis of anti-inflammatory activity.

Materials and methods

The in vitro anti-inflammatory activity of methanol extract of liquid cultured mycelia of Antrodia cinnamomea (MEMAC) was judged by the measurement of the produced levels of pro-inflammatory cytokines and mediators in lipopolysaccharide (LPS)-stimulated RAW264.7 cells and human peripheral blood mononuclear cells (PBMCs). The in vivo anti-inflammatory activity of MEMAC was evaluated using carrageenan-induced hind paw edema in mice, the activities of catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) in the liver and the levels of malondialdehyde (MDA) and nitrite oxide (NO) in the edema paw. The levels of serum NO and TNF-α were measured. The MEMAC was administered at the concentrations of 100, 200, and 400 mg/kg body weight of mouse.

Results

MEMAC inhibited the production of LPS-induced pro-inflammatory cytokines (TNF-α and IL-6) and mediators (NO and PGE2) in RAW264.7 cells and human PBMCs. Data from Western blotting showed that MEMAC decreased the levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) protein expression in LPS-stimulated RAW264.7 macrophages. In vivo, MEMAC showed significant (p < 0.05) anti-inflammatory activity by reducing the edema volume in carrageenan-induced paw edema in mice. MEMAC (400 mg/kg) also reduced the carrageenan-induced leukocyte migration (50.92 ± 5.71%). Further, MEMAC increased the activities of CAT, SOD, and GPx in the liver tissue and decreased the levels of serum NO and TNF-α after carrageenan administration.

Conclusions

Our results showed that MEMAC has the anti-inflammatory property both in vitro and in vivo, suggesting that it may be a potential preventive or therapeutic candidate for the treatment of inflammatory disorders.     

Ganoderma lucidum induced apoptosis in NB4 human leukemia cells: Involvement of Akt and Erk

Aim of the study

The final goal of this work was to study the toxic and apoptosis effects induced by fractions from Ganoderma lucidum [Ganoderma lucidum (Curtis) P. Karst.; Ganodermataceae Donk] on NB4 human leukemia cells.

Materials and methods

Two aqueous extracts and a methanol-extracted column-chromatography semipurified fraction were obtained from Ganoderma lucidum fruiting body. Flow cytometry analyses were used to measure cell viability, cell cycle and DNA fragmentation and to quantify apoptosis. Western-blot analyses were used to quantify changes in apoptosis proteins and intracellular kinases.

Results

Aqueous extracts slightly reduce cell viability and induce DNA fragmentation in NB4 cells. Methanol-extracted semipurified fraction at dilutions down to 15% or 40% of the initial fraction concentration reduced significantly the viability of these leukemia cells (treated for 19 h) with induction of DNA fragmentation and induction of apoptosis. Overmore, the dilution down to 15% of the initial E3 concentration induced a reduction of p53 levels, of the Bcl2/Bax relationship as well as reduced levels of both unphosphorylated and phosphorylated Akt (Protein kinase Akt, protein kinase B) and Erk (Erk1 and 2).

Conclusions

Induction of apoptosis and alterations in signal transduction kinases (Akt and Erk) are produced by active fractions from Ganoderma lucidum on human leukemia cells. These data could be of important relevance from the viewpoint of antitumor actions of compounds from Ganoderma lucidum. Eventual therapy applications in leukemia cells might be developed.     

Evaluation of anti-migraine potential of Areca catechu to prevent nitroglycerin-induced delayed inflammation in rat meninges: possible involvement of NOS inhibition

Ethnopharmacological relevance

Areca catechu nut extract is a popular folk remedy for the treatment of migraine in Kerala and Tamil Nadu states of India.

Aim of the study

In order to prove the claimed utilization of plant, the effect of hydroalcoholic extract of Areca catechu nut (ANE) was investigated in nitroglycerine induced inflammation in rat meninges. In these models infusion of nitric oxide donor glyceryl trinitrate (GTN) produces augmented plasma protein extravasation (PPE) in dura mater, provides an important substrate for the development of migraine in rats.

Materials and methods

The effect on plasma protein extravasation was assessed in both the models of intravenous and topical GTN application following oral administration of ANE (250 mg/kg and 500 mg/kg) in both curative and preventive treatment and compared with that of control positive. The l-NAME (15 mg/kg, i.v.) was used as reference standard. Plasma protein extravasation was measured using fluorescein as marker and was measured using a Perkin-Elmer LS-30 luminescence spectrometer.

Results

Expression of iNOS in the spleen after intravenous injection produced PPE into the dura mater in control positive group was significantly (P < 0.01) reduced to 1.553 ± 0.02499 and 1.398 ± 0.01887 by preventive treatment with ANE at the dose of 250 and 500 mg/kg, orally, respectively. The extravasation produced by topical GTN due to expression of iNOS in dural macrophages was also reduced to 1.555 ± 0.03384 and 1.425 ± 0.01204 by preventive treatment with ANE at the dose of 250 and 500 mg/kg, orally, respectively. While ANE do not showed any significant results in curative treatment in both the models of i.v. and topical GTN application.

Conclusion

These findings collectively indicate that the extract exhibited significant inhibition of iNOS, which may be the probable mechanism for its anti-migraine activity, providing evidence, at least in part, for its folkloric use.     

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.     

Sida rhomboidea.Roxb leaf extract ameliorates gentamicin induced nephrotoxicity and renal dysfunction in rats

Ethnopharmacological relevance

Sida rhomboidea.Roxb (SR) known as “Mahabala” in Ayurveda and marketed as “Shahadeyi” is used in ethnomedicine to treat ailments such as dysuria and urinary disorders.

Aim of the study

To evaluate nephroprotective potential of SR against gentamicin (GM) induced nephrotoxicity and renal dysfunction.

Materials and methods

Nephrotoxicity was induced in rats with GM (100 mg/kg bodyweight (i.p.) for 8 days) and were treated with SR extract (200 and 400 mg/kg bodyweight (p.o.) for 8 days) or 0.5% carboxymethyl cellulose (vehicle). Plasma and urine urea and creatinine, renal enzymatic and non-enzymatic antioxidants along with lipid peroxidation were evaluated in various experimental groups.

Results

GM treatment induced significant elevation (p < 0.05) in plasma and urine urea, creatinine, renal lipid peroxidation along with significant decrement (p < 0.05) in renal enzymatic and non-enzymatic antioxidants. SR treatment to GM treated rats (GM + SR) recorded significant decrement (p < 0.05) in plasma and urine urea and creatinine, renal lipid peroxidation along with significant increment (p < 0.05) in renal enzymatic and non-enzymatic antioxidants.

Conclusion

SR leaf extract ameliorates GM induced nephrotoxicity and renal dysfunction and thus validates its ethnomedicinal use.     

Hypnotic effect of jujubosides from Semen Ziziphi Spinosae

Ethnopharmacological significance

Semen Ziziphi Spinosae, the seed of Ziziphus jujuba Mill. var. spinosa (bunge) Hu ex H.F. Chow has been widely used in treating insomnia and anxiety.

Aim of the study

In this study, we investigated the hypnotic effect of jujubosides, one of the major components (saponin) of Semen Ziziphi Spinosae, in both day and night period.

Materials and methods

After the administration of jujubosides (9 mg/kg once per day for 3 days), sleep parameters were examined by EEG and EMG analysis in normal rats and the action of jujubosides on pentobarbital-induced sleep assessed by the loss-of righting reflex.

Results

During daytime (9:00–15:00), jujubosides significantly increased the total sleep and rapid eye movement (REM) sleep without significant influence on non-REM (NREM) sleep. During nighttime (21:00–3:00), jujubosides significantly increased the total sleep and NREM sleep especially the light sleep while showed no significant effect on REM sleep and slow wave sleep (SWS). In pentobarbital-treated mice, jujubosides significantly augmented the hypnotic effect of pentobarbital (45 mg/kg, i.p.), proved by increasing sleep time and this augmentative effect was potentiated by 5-hydroxytryptophan (2 mg/kg, i.p.). Furthermore, jujubosides inhibited the para-chlorophenylalanine-induced suppression of pentobarbital-induced hypnosis.

Conclusions

These results suggested that the hypnotic effect of jujubosides on normal rats may be influenced by circadian rhythm and the serotonergic system may involve in the hypnotic effect of jujubosides. Jujubosides may be good source of lead compounds for novel hypnotics.     

In vivo hepatoprotective activity of the aqueous extract of Artemisia absinthium L. against chemically and immunologically induced liver injuries in mice

Aim of the study

This study aimed to evaluate in vivo hepatoprotective activity of the aqueous extract of Artemisia absinthium L. (AEAA), which has been used for the treatment of liver disorders in Traditional Uighur Medicine.

Materials and methods

Qualitative and quantitative phytochemical analysis of the AEAA was performed by means of thin layer chromatography and spectrophometric assays. Aqueous extract (50, 100 or 200 mg/kg body weight/day) was administered orally to experimental mice. Liver injury was induced chemically, by a single CCl₄ administration (0.1% in olive oil, 10 ml/kg, i.v.), or immunologically, by injection of endotoxin (LPS, 10 μg, i.v.) in BCG-primed mice. The levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), tumor necrosis factor-α (TNF-α) and interleukin-1 (IL-1) in mouse sera, as well as superoxide dismutase (SOD), glutathione peroxidase (GPx) and malondialdehyde (MDA) in mouse liver tissues were measured. The biochemical observations were supplemented by histopathological examination.

Results

Obtained results demonstrated that the pretreatment with AEAA significantly (P < 0.001) and dose-dependently prevented chemically or immunologically induced increase in serum levels of hepatic enzymes. Furthermore, AEAA significantly (P < 0.05) reduced the lipid peroxidation in the liver tissue and restored activities of defense antioxidant enzymes SOD and GPx towards normal levels. In the BCG/LPS model, increase of the levels of important pro-inflammatory mediators TNF-α and IL-1 was significantly (P < 0.01) suppressed by AEAA pretreatment. Histopathology of the liver tissue showed that AEAA attenuated the hepatocellular necrosis and led to reduction of inflammatory cells infiltration. Phytochemical analyses revealed the presence of sesquiterpene lactones, flavonoids, phenolic acids and tannins in the AEAA.

Conclusions

The results of this study strongly indicate the protective efect of AEAA against acute liver injury which may be attributed to its antioxidative and/or immunomodulatory activity, and thereby scientifically support its traditional use.     

Anti-inflammatory effects of Pulvis Fellis Suis extract in mice with ulcerative colitis

Ethnopharmacological relevance

Pulvis Fellis Suis is used in folk medicines to treat intestinal diseases, acute pharyngitis, whooping cough and asthma in China. Although several reports indicate that Pulvis Fellis Suis display diverse biological activities, such as antibacterial, anti-inflammatory and anti-infusorian effects, its effects on ulcerative colitis have not been previously explored.

Aim of the study

The purpose of the present study is to assess the anti-inflammatory effect of Pulvis Fellis Suis (PFS) extract in acute ulcerative colitis model induced by trinitrobenzene sulfonic acid (TNBS) in mice.

Materials and methods

Different doses of Pulvis Fellis Suis extract (100, 200 and 400 mg/kg/day) and sulfasalazine (500 mg/kg/day) were administered by gavage for 7 days after the induction of colitis with TNBS. The efficacy of PFS was studied by macroscopical and histological scoring systems as well as myeloperoxidase (MPO) activity. Serum levels, including tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were assayed by enzyme-linked immunoassay. The expression of cyclooxygenase (COX)-2 in the colons was assessed by immunohistochemical analysis.

Results

Treatment with PFS significantly attenuated macroscopic damage as compared with TNBS (P < 0.01). Histological analysis showed that PFS improved the microscopic structure and preserved some areas of the colonic mucosa structure. In addition, administration of PFS effectively inhibited COX-2 protein expression and MPO activity accumulation. TNF-α and IL-6 levels were also diminished dose-dependently (P < 0.05, P < 0.01), and IL-6 level obtained had no significant results by small dose of PFS. All the effects of these parameters were comparable to that of the standard sulfasalazine, especially at the highest dose level.

Conclusions

We have shown for the first time that PFS has an anti-inflammatory effect in TNBS-induced ulcerative colitis which might be related to the reduction of up-regulated TNF-α and IL-6 production, and that it may have therapeutic value in the setting of inflammatory bowel disease (IBD).     

Protective effects of methanolic extract of Hedyotis puberula (G. Don) R. Br. ex Arn. against experimentally induced gastric ulcers in rat

Aim of the study

This study was aimed to evaluate the antiulcer activity of the whole plants of Hedyotis puberula (G. Don) R. Br. ex Arn.

Materials and methods

Gastroprotective potential of the Hedyotis puberula methanol extract (200 and 400 mg/kg body weight) was studied on indomethacin (IND), ethanol and pyloric ligation (PL)-induced gastric ulcer models in rats.

Results

The treatment with Hedyotis puberula extract at 400 mg/kg p.o. protected the rats against the ulceration which was comparable to the reference drug omeprazole. Pretreatment with extract protected rats from gastric lesion development by way of increased pH and decreased volume, acidity and pepsin content of gastric secretion. Furthermore, total carbohydrate: protein ratio of the gastric juice were noticeably increased in pretreated rats.

Conclusion

Results of our study showed that Hedyotis puberula possess significant gastroprotective activity and validate the folklore claim.     

Identification and pharmacological characterization of the anti-inflammatory principal of the leaves of dwarf elder (Sambucus ebulus L.)

Aim of the study

The performed investigations aimed on the identification of the anti-inflammatory principal of extracts of leaves of Sambucus ebulus L. (dwarf elder) in order to rationalize the traditional use of this plant for the treatment of chronically inflammatory diseases.

Materials and methods

Dwarf elder leaf extract was subjected to activity guided fractionation using inhibition of TNFα induced expression of vascular cell adhesion molecule 1 (VCAM-1) on the surface of human umbilical vein endothelial cells (HUVECs) as monitoring tool (positive control: parthenolide 10 μM, VCAM-1 expression (% of control): 5.35 ± 0.38%).

Results

Bio-guided isolation resulted in identification of ursolic acid as anti-inflammatory principal. Besides its inhibitory effects against TNFα induced expression of VCAM-1 (IC₅₀ 6.25 μM), ursolic acid inhibits also TNFα induced expression of ICAM-1 (IC₅₀ value between 3.13 and 6.25 μM) (positive control: parthenolide 10 μM, ICAM-1 expression (% of control): 38.89 ± 16.6%). Toxic effects of ursolic acid on HUVECs can be drastically reduced using an enriched extract instead of the pure compound.

Conclusions

Our findings suggest an additional mechanism of the anti-inflammatory activity of ursolic acid by demonstrating its ability to inhibit TNFα-stimulated expression of VCAM-1 and ICAM-1 and support the traditional use of extracts and preparations of Sambucus ebulus L., rich in ursolic acid, for the treatment of chronically inflammatory processes.     

Anti-inflammatory activities of Chinese herbal medicine sinomenine and Liang Miao San on tumor necrosis factor-α-activated human fibroblast-like synoviocytes in rheumatoid arthritis

Aim of the study

Sinomenine, an alkaloid isolated from the root of Sinomenium acutum, has been used to alleviate the symptoms of rheumatic diseases. Liang Miao San (LMS), composed of the herbs Rhizoma Atractylodis (Cangzhu) and Cotex Phellodendri (Huangbai), is another traditional Chinese medicine formula for rheumatoid arthritis (RA) treatment. Although numerous studies have demonstrated the potential anti-inflammatory activities of sinomenine and LMS, the underlying intracellular mechanisms regulating the anti-inflammatory activities of sinomenine and LMS on human primary fibroblast-like synoviocytes (FLS) from RA patients and normal control subjects have not been elucidated.

Materials and methods

We investigated the in vitro anti-inflammatory activity of sinomenine and LMS on inflammatory cytokine tumor necrosis factor (TNF)-α-mediated activation of human normal and RA-FLS. The underlying intracellular signaling molecules were analyzed quantitatively using flow cytometry.

Results

Sinomenine was found to significantly inhibit TNF-α induced cell surface expression of vascular cell adhesion molecule (VCAM)-1 and release of inflammatory cytokine and chemokine IL-6, CCL2 and CXCL8 from both normal and RA-FLS (all p < 0.05). Moreover, the suppression of sinomenine on TNF-α induced VCAM-1 expression and IL-6 release of RA-FLS was significantly higher than that of normal FLS (p < 0.05). LMS significantly inhibited TNF-α-induced inflammatory chemokines CXCL10 and CCL5 release from both normal and RA-FLS, with significantly higher suppression on CXCL10 secretion in RA-FLS than that of normal FLS (all p < 0.05). Further investigations showed that sinomenine and LMS could significantly suppress TNF-α-induced phosphorylation of inhibitor κBα and extracellular signal-regulated protein kinase, the central signaling molecules mediating TNF-α-induced VCAM-1 expression and chemokine production.

Conclusion

Our results therefore provide a new insight into the differential anti-inflammatory activities of sinomenine and LMS through the suppression of TNF-α-activated FLS by modulating distinct intracellular signaling pathways in RA.     

Medicinal potions used against infectious bowel diseases in Mayan traditional medicine

Aim of the study

Since one of the main health problems of the indigenous population are infectious bowel diseases, we decided to test Mayan medicinal potions used to treat these conditions against some of the causal agents.

Materials and methods

Thirty-one herbal formulations used for the treatment of infectious bowel diseases were prepared according to the collected ethnobotanical data. Their activities were tested against some of the causal agents of diarrheic symptoms, such as Entamoeba histolytica, Giardia lamblia, Escherichia coli, Klebsiella pneumoniae, Salmonella typhi and Shigella flexneri.

Results

Nine formulations were active against bacteria (MIC = 0.5 mg/ml), four on Entamoeba histolytica, and seven on Giardia lamblia (IC₅₀ ≤ 20 μg/ml).

Conclusions

This work supports the use of the traditional Mayan formulations against some infectious bowel diseases, and it is the first step towards their study.