Potential ex vivo immunomodulatory effects of San-Huang-Xie-Xin-Tang and its component herbs on mice and humans

Aim of the study

San-Huang-Xie-Xin-Tang (SHXXT), an important Chinese medicine formula, contains Rhei Rhizoma (RR), Scutellariae Radix (SR) and Coptidis Rhizoma (CR). RR and SR are abundant in anthraquinone and flavonoid polyphenols. Pharmacokinetic study of SHXXT indicated that glucuronides were the predominant forms of polyphenols in rats.

Materials and methods

As an extension of pharmacokinetic study, the serum metabolites of SHXXT, RR, SR and CR were prepared from rats and quantitated, then the immunomodulation effects were examined by culturing these serum metabolites with murine and human immune cells.

Results

The results indicated that the inhibitions on nitric oxide (NO) and cytokine production from mitogen-activated peritoneal macrophages by the serum metabolites of SHXXT, RR, SR and CR were through reducing the protein expression of inducible NO synthase (iNOS) and the IC₅₀ were 0.8%, 1.5%, 3.0% and 0.8% of their blood concentrations, respectively. In addition, the serum metabolites of SHXXT, RR, SR and CR significantly decreased the ratios of interferon-gamma (IFN-γ) to interleukin (IL)-4 in mitogen-stimulated mice spleen cells and human peripheral blood mononuclear cells (PBMCs). Moreover, the serum metabolites of SHXXT and SR significantly arrested the mitogen-stimulated mice spleen cells at G2/M stage.

Conclusions

In conclusion, the serum metabolites of SHXXT and the component herbs exerted promising modulation activities on the immune functions and the cell cycle distribution of mice and human immune cells. We suggest that SHXXT is a promising remedy for immunomodulation through Th1/Th2 regulation.     

The effects of Rhizoma drynariae on interleukin-2 and T-lymphocyte levels in rats after severe head injury

Objective

To investigate the effects of Rhizoma drynariae on the levels of interleukin-2(IL-2) and T-lymphocyte subset in rats with severe head injury (SHI).

Methods

72 Sprague Dawley (SD) rats were randomly divided into 3 groups: control group, model group, and R. drynariae group. The experimental group received intragastrical infusions of with aqueous R. drynariae extract four hours after SHI while the other groups were administered with equivalent volumes of physiological saline. Infusions were administered to the 3 groups once a day for 7 d. IL-2 and T-lymphocyte (CD3, CD4, CD8) levels were measured at 24, 72, and 168 h after initial infusion.

Results

The levels of IL-2 and CD4 T cells reduced obviously after 24 h in the model group (P<0.05), but recovered to the levels of the control group after 72 h, and remained elevated after 168 h. In the R. drynariae group, IL-2 and CD4 levels were did not decrease while the level of CD8 T cells was reduced significantly (P<0.05).

Conclusions

R. drynariae can protect against immune dysfunction or improve immunity in rats with severe head injury (SHI).     

Th1/Th2 cytokines’ expression and production by propolis-treated mice

Aim of the study

Propolis is a natural product extensively used in food and beverages to improve health and to prevent diseases, showing immunomodulatory properties. The goal of this work was to evaluate the effect of propolis administration over a short-term to mice on Th1 (IL-2 and IFN-γ) and Th2 (IL-4 and IL-10) cytokines’ expression and production.

Materials and methods

Propolis was administered for 3 days to mice by gavage, spleens were removed and RNA was extracted to assess cytokines’ expression by real-time PCR. Supernatants of spleen cell cultures were used for cytokines determination by ELISA.

Results

Propolis administration to mice did not affect IL-2, IL-4 and IL-10 expression and production, while IFN-γ production was inhibited in the splenocyte cultures stimulated or not by Con A.

Conclusions

Since IFN-γ is a pro-inflammatory cytokine, our data suggest that propolis administration over a short-term to mice may be associated with anti-inflammatory effects in vivo, and further assays could check propolis efficiency in inflammatory diseases.     

Effects of Catalpa ovata stem bark on atopic dermatitis-like skin lesions in NC/Nga mice

Ethnopharmacological relevance

The stem bark of Catalpa ovata has been used as a traditional herbal medicine for the treatment of various inflammatory diseases such as itching and scabies.

Aim of the study

In the present study, we investigated the anti-AD effects of Catalpa ovata stem bark on Dermatophagoides farinae-induced AD in a NC/Nga mouse AD model. We determined dermatitis score, histology, IgE, cytokines, and chemokines related to hypersensitive immune responses in AD. The mechanism of action was also investigated using HaCaT cells.

Materials and methods

We investigated the topical effects of Catalpa ovata stem bark on AD-like skin lesions in NC/Nga mice. Five category-experiments were performed, including assessment of dermatitis score; histological analysis of dorsal skin lesions; quantitative measurement of serum total IgE; quantitative measurement of cytokines (IL-1β, IL-4, IL-5, IL-6, IL-13, TNF-α) from dorsal tissue; and RT-PCR analysis of for TSLP and TARC mRNA expression in HaCaT cells.

Results

The clinical dermatitis score was significantly lower in Catalpa ovata extract (COE) groups than in the control group. Histological analysis showed that COE inhibited hypertrophy and hyperkeratosis of the epidermis, intracellular edema, and reduced the infiltration of inflammatory cells. COE significantly inhibited serum total IgE; Th2 cytokines IL-4, IL-5 and IL-13; pro-inflammatory cytokines IL-1β, IL6 and TNF-α; the Th2 chemokine TARC and the pro-Th2 cytokine TSLP.

Conclusion

These results demonstrate that Catalpa ovata stem bark may be a useful external medicine for treatment of AD. Further investigation is necessary to determine appropriate COE dosage and to evaluate the safety of this medicinal herb.     

Augmentation of expression of immunocytes’ functions by seed extract of Ziziphus mauritiana (Lamk.)

Aim of the study

The present study was carried out to evaluate the immunomodulatory potential of Ziziphus mauritiana (Lamk.) seed extract to ascertain the folkloric claim as immunomodulator.

Materials and methods

The aqueous-ethanolic seed extract (100–400 mg kg⁻¹) of Z. mauritiana was investigated for immunomodulatory potential in mice. The extract was standardized with HPLC using betulinic acid as a marker. Functions of various immunocytes in the form of humoral (development of anti-SRBC (sheep red blood cells) antibody titers) and cell-mediated immune response (delayed type hypersensitivity, nitroblue tetrazolium reduction, inducible nitric oxide synthase activity and bactericidal activity) was studied in SRBC immunized mice. The cytokine, IFN-γ (interferon-gamma) and IL-4 (interleukin-4) secretion was also measured quantitatively by ELISA as the expression of functions of Th-1 and Th-2 respectively. Levamisole (2.5 mg kg⁻¹) was used as standard drug.

Results

The seed extract demonstrated significant (P < 0.05–0.001) up-regulation of cell-mediated, humoral immune response and Th-1 mediated cytokine IFN-γ and decline in Th-2 mediated cytokine IL-4. At higher dose of extract the results were comparable to that of the levamisole.

Conclusion

The immunostimulatory potential of this seed extract is likely to be mediated through its effect on macrophage function and Th-1 mediated immunity confirming the folkloric use of this plant.     

Inhibitory effects of the root extract of Litsea cubeba (lour.) pers. on adjuvant arthritis in rats

Ethnopharmacological relevance

The dried root of Litsea cubeba (Lour.) Pers. (Family Lauraceae) has long been used as a folk remedy in Traditional Chinese Medicine (TCM) and Dai Ethnopharmacy for the treatment of rheumatic diseases in southwestern China.

Aim of the study

This study investigated the preventive efficacy of Litsea cubeba root in treating rheumatoid arthritis using Freund's complete adjuvant (CFA) induced arthritis (AA) in rat model.

Materials and methods

Arthritis was induced in male Wistar rats by immunization with CFA. Ethanol extract (EELC) and water extract (WELC) of Litsea cubeba root both at 50 mg/kg and 200 mg/kg were orally administered from a day after the induction of arthritis. Paw swelling, arthritic score, body weight growth rate, index of thymus and spleen were observed, and the production of TNF-α, IL-1β, IL-6 and IL-10 in serum were measured by enzyme-linked immunosorbent assay. The expression levels of inflammatory enzymes like cyclooxygenase and lipoxygenase were also measured by enzyme-linked immunosorbent assay. Moreover, histological changes in the ankle joint were analyzed in AA rats.

Results

Both EELC and WELC significantly suppressed paw swelling and arthritic score, increased the loss in body weight and decreased the index of thymus. Histopathological improvement in joint architecture was also observed in EELC, WELC-treated AA rats. The expression levels of cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX) were decreased on treatment with EELC and WELC. Furthermore, the overproduction of TNF-α, IL-1β and IL-6 were remarkably attenuated in serum of all Litsea cubeba-treated rats, however, IL-10 was markedly increased at doses of 50 mg/kg of EELC and WELC.

Conclusions

These results indicate that extract of Litsea cubeba root significantly attenuates adjuvant arthritis in rats by decreasing the levels of TNF-α, IL-1β and IL-6 and increasing of IL-10 in serum as well as down-regulate the levels of inflammatory enzymes such as COX-2 and 5-LOX. This suggests that Litsea cubeba root might be used as a therapeutic agent for the treatment of human arthritis.     

Huo-Luo-Xiao-Ling Dan modulates antigen-directed immune response in adjuvant-induced inflammation

Ethnopharmacological relevance

HLXL is a traditional Chinese medicine that has long been used in folk medicine for the treatment of chronic inflammatory diseases. However, the precise immunological mechanisms by which HLXL mediates its anti-inflammatory activity are not fully defined.

Aim of the study

To determine the effects of HLXL on antigen-specific immune parameters in adjuvant-induced inflammation model in the Lewis rat.

Materials and methods

Rats were fed daily with either HLXL (2.3 g/kg) or vehicle (water) beginning 3 days before subcutaneous injection of heat-killed Mycobacterium tuberculosis H37Ra (Mtb), and then continued for another 6 days. After 9 days of Mtb injection, the draining lymph node cells were tested for T cell proliferative and cytokine responses against mycobacterial heat-shock protein 65 (Bhsp65). Moreover, sera were tested for anti-Bhsp65 antibodies and nitric oxide (NO).

Results

HLXL-treated rats showed reduced T cell proliferative response to Bhsp65 compared to control rats. Furthermore, HLXL suppressed IL-17 response but enhanced IL-10 response without much effect on IFN-γ. HLXL treatment also reduced the levels of anti-Bhsp65 antibodies but not that of NO.

Conclusions

HLXL feeding modulated both the cellular and the humoral immune response to Bhsp65 favoring an anti-inflammatory milieu for the suppression of adjuvant-induced inflammation.     

Experimental study on anti-inflammatory activity of a TCM recipe consisting of the supercritical fluid CO2 extract of Chrysanthemum indicum, Patchouli Oil and Zedoary Turmeric Oil in vivo

Ethnopharmacological relevance

Chrysanthemum indicum (Compositae) Linné, Pogostemon cablin (Blanco) Benth and Curcuma wenyujin (Zingiberaceae) Y. H. Chen et C. Ling are three of the extensively used herbal remedies among traditional Chinese medicines for the purpose of anti-inflammation. A traditional Chinese medicine (TCM) recipe named CPZ consisting extracts of the above three herbs, has shown noteworthy anti-influenza activity, which is closely related to its anti-inflammatory feature.

Aim of this study

To investigated the anti-inflammtory activity of CPZ in vivo for a further exploration of the recipe's anti-inflammatory properties.

Materials and methods

The anti-inflammatory property of CPZ on acute inflammation was evaluated by inflammatory models of dimethylbenzene (DMB)-induced ear vasodilatation and acetic acid-induced capillary permeability enhancement in mice, as well as the carrageenan-induced paw edema rat model, in which inflammation-related cytokine including prostaglandin E₂ (PGE₂), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and nitric oxide (NO) in the edematous paw tissue were determined by enzyme linked immunosorbent assay (ELISA). Moreover, effect of CPZ on chronic inflammation was observed through granuloma formation in rats subjected to cotton pellet implantation.

Results

CPZ (340, 170, and 85 mg/kg for mice, p.o.) not only decreased the DMB-induced ear vasodilatation but also attenuated capillary permeability under acetic acid challenge in mice. And the significant inhibition on carrageenan-induced paw edema was observed. Further more, the ELISA results showed that CPZ (170, 85, and 42.5 mg/kg for rats, p.o.) could up-regulate the level of IL-1β in the edema paw tissue of rats significantly while down-regulate that of PGE₂, but no apparent effect on TNF-α or NO was observed in the test. Besides, CPZ had a certain degree of restraining effect on the cotton pellet-induced granuloma formation in rats and the highest dose of 170 mg/kg even showed a significant suppression on it.

Conclusion

The above results indicated that CPZ possessed a potent anti-inflammatory activity, which is indicated to be closely associated with its regulation on IL-1β and PGE₂ thereby mediating the inflammatory response acting at an appropriate level.     

Comparisons on enhancing the immunity of fresh and dry Cordyceps militaris in vivo and in vitro

Ethnopharmacological relevance

The immunomodulatory capacities of fresh Cordyceps militaris (FCM) and dry Cordyceps militaris (DCM) were compared.

Materials and methods

In vivo immunomodulatory assay, different doses of FCM and DCM were orally administrated over a period of 15 days in a cyclophosphamide (CY) induced immunosuppression mice; in vitro testing, the spleen cells were extracted from healthy mice and treaded with CY, then cultured with different dose of FCM or DCM; the contents of Cordyceps militaris polysaccharide (CMP), cordycepin, adenosine, total polyphenol (TP) and total flavonoids (TF) in FCM and DCM were measured.

Results

Our studys indicated that, FCM was significantly stronger than DCM on increasing the spleen and thymus indexes, spleen lymphocyte activity, macrophage function, and promoting the levels of IL-2, IFN-γ in vivo and in vitro. The contents of immunomodulatory CMP and TF in FCM were markedly higher than in DCM.

Conclusion

All these results suggested that FCM was superior to DCM on enhancing immunity.     

Scorpio and Scolopendra attenuate inflammation and articular damage in rats with collagen-induced arthritis

Ethnopharmacological relevance

Scorpio and Scolopendra (SS) are two traditional Chinese medicines, which are generally used to treat rheumatoid arthritis (RA) in China. However, the mechanism is so far unclear. The purpose of this study was to explore the effects and mechanisms of SS in attenuating inflammation and joint injury in collagen-induced arthritis in rats.

Materials and methods

RA was induced in Wistar rats by injection of collagen, meanwhile, the rats were administrated daily either SS (0.4 g/kg, 0.2 g/kg, and 0.1 g/kg) or vehicle (physiological saline) for 42 days. The therapeutic effect of SS on RA was evaluated by pathological methods. T lymphocyte subsets and anti-collagen type II (CII) antibody were tested in peripheral blood. Tumor necrosis factor alpha (TNF-α), interleukin-1β (IL-1β), interleukin-4 (IL-4) and interleukin-10 (IL-10) were assessed in tissue homogenate of fresh joints.

Results

The inflammation and articular damage in SS powder-treated rats were attenuated significantly. In addition, SS powder was revealed to modulate the equilibrium of T lymphocyte subsets, down-regulate TNF-α and IL-1β, up-regulate IL-4 and IL-10, and significantly suppress the level of anti-CII antibody.

Conclusions

Scorpio and Scolopendra, when used as a combination, reveal desirable effect for treatment of rheumatoid arthritis, and this beneficial effect may be accomplished through normalization of T lymphocyte subsets and the balance of Th1/Th2 cytokines.     

Effects of Bambusae caulis in Liquamen on the development of atopic dermatitis-like skin lesions in hairless mice

Ethnopharmacological relevance

Bambusae caulis in Liquamen (BCL) is a nutritious liquid extracted from heat-treated fresh bamboo stems. It is an important traditional herbal medicine used to treat coughs and asthma in East Asia. In recent years, it has been studied for its anti-inflammatory, anti-allergenic, and immune-regulating properties.

Aim of the study

To examine whether BCL suppresses the development of 2,4-dinitrochlorobenzene (DNCB)-induced atopic dermatitis (AD)-like skin lesions in hairless mice.

Materials and methods

The effects of BCL were analyzed by measuring transepidermal water loss (TEWL), melanin content, and erythema in the skin, leukocyte numbers and IgE levels in the serum, and mRNA expression of relevant cytokines in the spleen.

Results

The transdermal administration of BCL to hairless mice inhibited the development of DNCB-induced AD-like skin lesions by suppressing TEWL, melanin production and erythema of skin, the number of leukocytes and the level of IgE in serum, and the mRNA expression of IL-4, IL-13, and TNF-α in the spleen. However, BCL administration increased the expression of IFN-γ in the spleen.

Conclusions

These findings indicate that BCL suppresses the development of DNCB-induced AD-like skin lesions in hairless mice, suggesting that BCL may be a potential therapeutic agent for AD in a clinical setting.     

Anti-inflammatory activity of ethyl acetate fraction of the seeds of Brucea Javanica

Ethnopharmacological relevance

The seeds of Brucea javanica (L.) Merr. (Yadanzi in Chinese) have been used for the treatment of inflammation, dysentery, malaria, and cancer in Chinese traditional medicine. However, the anti-inflammatory mechanism of Brucea javanica has not been fully elucidated. This study examined the anti-inflammatory activity of ethyl acetate fraction of the seeds of Brucea javanica (EA-BJ) in vitro and in vivo.

Materials and methods

The anti-inflammatory activity of EA-BJ and its ability to modulate the production of NO, PGE2, TNF-α, IL-1β, IL-6 and IL-10 inflammatory mediators in lipopolysaccharide-activated RAW 264.7 macrophage were evaluated. Moreover, the anti-inflammatory activity of EA-BJ was also in vivo assayed by carrageenan induced paw edema in mice.

Results

In vitro assays showed remarkable anti-inflammatory activity of EA-BJ, through the inhibition of production of NO, PGE2, TNF-α, IL-1β and IL-6 inflammatory mediators and induction of production of IL-10 anti-inflammatory cytokine. In vivo assays showed anti-inflammatory activity for decrement of the paw edema in carrageenan induced paw edema test.

Conclusion

The results obtained in vitro and in vivo showed that possible anti-inflammatory effects of EA-BJ may be attributed to inhibition pro-inflammatory mediators production, NO, PGE2, TNF-α, IL-1β and IL-6 and to increase production of IL-10 anti-inflammatory cytokine. The seeds of Brucea javanica may thus prove beneficial in the treatment of inflammatory diseases.     

Attenuation of airway hyperreactivity and T helper cell type 2 responses by coumarins from Peucedanum praeruptorum Dunn in a murine model of allergic airway inflammation

Ethnopharmacological relevance

The root of Peucedanum praeruptorum Dunn (PPD) is a commonly used traditional Chinese medicine for the treatment of asthma. Its major constituents, coumarins, were presumed to be responsible for its efficacy.

Aim of the study

The potential of coumarins from PPD (CPPD) as anti-asthma agent was investigated.

Materials and methods

Female BALB/c mice were sensitized and challenged with ovalbumin (OVA) to induce allergic airway inflammation. CPPD was administered intragastrically before every OVA challenge. Airway reactivity to the intravenous administration of acetylcholine chloride was measured 48 h after final OVA inhalation. Airway inflammation was evaluated by leukocyte counts of bronchoalveolar lavage fluid (BALF) and histopathological analysis of lung lesions. Levels of interleukin (IL)-4, IL-5, IL-10, IL-13 and IFN-γ in BALF and OVA-specific immunoglobulin (Ig) E in serum, and activity of eosinophil peroxidase (EPO) in lung was measured. The percentage of CD4⁺CD25⁺Foxp3⁺ regulatory T cells among CD4⁺ T cells in spleen was analyzed by flow cytometry.

Results

Compared with model group, CPPD significantly reduced airway hyperreactivity and airway eosinophilic inflammation, improved pathologic lesion of the lungs, reduced levels of IL-4, IL-5, IL-13 in BALF and OVA-specific IgE in serum, inhibited the activities of EPO in lung, and up-regulated levels of IL-10 and IFN-γ in BALF as well as the percentage of CD4⁺CD25⁺Foxp3⁺ regulatory T cells in spleen.

Conclusion

CPPD can significantly suppress OVA-induced airway inflammation, airway hyperreactivity and Th2 predominant response in mice, showing great therapeutic potential for the treatment of allergic asthma.     

Anti-arthritic activity of Xanthium strumarium L. extract on complete Freund׳s adjuvant induced arthritis in rats

Ethnopharmacological relevance

Xanthium strumarium L. fruit (Xanthiu fruit) has been traditionally used as a medicinal herb in China for the treatment of many ailments including rheumatoid arthritis. However, the anti-arthritic activity of Xanthium strumarium fruit has still not been demonstrated. In the present study, we confirmed that the extract of Xanthium strumarium (EXS) prevents rheumatoid arthritis induced by Complete Freund?s Adjuvant (CFA) in rats.

Materials and methods

Male Wistar rats (160±10 g) were immunized by intradermal injection of 0.1 mL of CFA into the left hind metatarsal footpad. EXS was administered orally at a dose of 300 and 75 mg/kg once a day after the induction of adjuvant arthritis. Methotrexate (3 mg/kg, twice a week) was used as a positive control. Paw swelling, arthritic score, body weight loss, spleen index, thymus index, serum cytokines, inflammatory mediators and histological change were measured. The chemical profile of EXS was analyzed by HPLC-DAD.

Results

We found that the EXS significantly suppressed paw swelling and arthritic score, increased body weight loss and decreased the thymus index. The overproduction of TNF-α and IL-1β were remarkably suppressed in the serum of all EXS-treated rats, and in contrast IL-10 was markedly increased. The level of COX-2 and 5-LOX was also decreased with EXS treatment. Ten phenolic acid derivatives were identified from 14 detected peaks by HPLC-DAD with the reference substances and verified by LC–MS.

Conclusions

These results suggest the potential effect of EXS as an anti-arthritis agent towards CFA-induced arthritis in rats. Xanthium strumarium has the potential to be regarded as a candidate for use in general therapeutics and as an immune-modulatory medicine in rheumatoid arthritis.     

Toxicity studies of Tithonia diversifolia A. Gray (Asteraceae) in rats

Objective

To investigate the toxicity of an ethanolic extract of the aerial parts of Tithonia diversifolia, used in Nigeria to treat malaria, in rats.

Materials and methods

A 70% ethanol extract was administered orally to adult Wistar rats at various dosages (400–1600 mg/kg) and the animals sacrificed and various organs examined at a range of times from 30 min up to 24 h after administration.

Results

The studies showed a dose- and time-dependent toxic effect, which was reversible on the kidney and liver while there was no noticeable adverse effect on the morphology of the heart, spleen and brain.

Conclusion

A 70% ethanol extract of the aerial parts of Tithonia diversifolia, which had previously been shown to reduce parasitemia in mice infected with Plasmodium, displayed kidney and liver toxicity at the lowest dose tested. The use of this plant extract against malaria therefore raises concerns over its safety.     

Serviceberry [Amelanchier alnifolia (Nutt.) Nutt. ex. M. Roem (Rosaceae)] leaf extract inhibits mammalian α-glucosidase activity and suppresses postprandial glycemic response in a mouse model of diet-induced obesity and hyperglycemia

Ethnopharmacological relevance

The use of medicinal plants for the treatment of diseases usually comes from the belief that they present low toxicity due their natural origin. However, it is necessary a toxicological and pharmacological evaluation for these plants. Tropaeolum majus is a medicinal plant used in popular medicine to treat several diseases, including cardiovascular disorders, urinary tract infections and asthma. Even though several studies proved its therapeutic effects, there are few toxicological studies with this species.

Aim of the study

The present study was carried out to evaluate the subchronic toxicity of the hydroethanolic extract obtained from leaves of T. majus (HETM) in Wistar rats.

Material and methods

Male and female Wistar rats received three doses of HETM (75, 375 and 750 mg/kg) for 28 days. After the treatments biochemical, hematological and histopathological parameters were analyzed.

Results

No significant alterations in the animal's body weight gain, relative organs weight, serum biochemical analysis, hematological or histopathological analyses of liver, kidneys and spleen were observed.

Conclusions

These results demonstrate the absence of subchronic toxicity due to oral treatment with HETM for 28 days in Wistar rats. However, other toxicological studies are necessary to evaluate the total safety of this plant.     

Intestinal anti-inflammatory activity of hydroalcoholic extracts of Phlomis purpurea L. and Phlomis lychnitis L. in the trinitrobenzenesulphonic acid model of rat colitis.

Ethnopharmacological relevance

Different species from genus Phlomis, frequently native from the the eastern Mediterranean zone, have been used in traditional medicine as an anti-inflammatory remedy. Among other constituents, they contain polyphenols that show antioxidant properties, which are interesting for the treatment of inflammatory pathologies associated with oxidative stress in humans, such as inflammatory bowel disease (IBD). The aim of this study was to evaluate the intestinal anti-inflammatoy effect of hydroalcoholic extracts of Phlomis lychnitis and P. purpurea in the trinitrobenzenesulphonic acid (TNBS) model of rat colitis, a well characterized experimental model with some resemblance to human IBD.

Materials and methods

Hydroalcoholic extracts of both plants were characterized by determining their polyphenolic content and then assayed in the TNBS model of rat colitis. For this purpose, female Wistar rats were assigned to seven groups (n=10): healthy control, untreated TNBS-colitis and five TNBS- colitis groups treated with Phlomis lychnitis (10 and 20 mg/kg), P. purpurea (10 and 25 mg/kg) and sulphasalazine (200 mg/kg), as a positive control. Treatments started the same day of TNBS colitis induction, and rats were sacrificed one week later. Colonic inflammation was evaluated both histologically and biochemically.

Results

The histological (macroscopic and microscopic) analysis of colonic samples revealed that both extracts showed an anti-inflammatory effect, which was confirmed biochemically by a decreased colonic MPO activity, a maker of neutrophil infiltration, an increased colonic glutathione content, which counteracts the oxidative status associated with the inflammatory process, and a down-regulated iNOS expression. However, only the extract of P. purpurea reduced the expression of the proinflammatory cytokines IL-1β and IL-17, the chemokines CINC-1 and MCP-1, as well as the adhesion molecule ICAM-1, ameliorating the altered immune response associated with the colonic inflammation. Furthermore, both P. lychnitis and P. purpurea extracts were able to significantly increase the expression of markers of epithelial integrity such as MUC-2, MUC-3 and villin, thus revealing an improvement in the altered colonic permeability that characterizes colonic inflammation.

Conclusions

Both extracts showed intestinal anti-inflammatory activity in the TNBS model of rat colitis, thus confirming their traditional use in digestive inflammatory complaints. In addition to their antioxidant properties, other mechanisms can contribute to this beneficial effect, like an improvement in the intestine epithelial barrier and a downregulation of the immune response.     

Ethanol extracts from Hemerocallis citrina attenuate the upregulation of proinflammatory cytokines and indoleamine 2,3-dioxygenase in rats

Ethnopharmacological relevance

Hemerocallis citrina, a traditional herbal medicine, has been used for the improvement of behavioral and emotional status in Eastern-Asia countries. Previous studies in our laboratory demonstrated that ethanol extracts from Hemerocallis citrina (HCE) enhanced monoamines and brain-derived neurotrophic factor (BDNF) in depression-like model of rodents.

Materials and methods

The present study extends earlier works on the role of anti-inflammation in regulating the antidepressant-like actions of HCE in rats exposed to chronic unpredictable mild stress (CUMS). Frontal cortex and hippocampal proinflammatory cytokines levels and indoleamine 2,3-dioxygenase (IDO) activity were measured after 4-week HCE treatment in the CUMS an control rats.

Results

Chronic administration of HCE reversed the decreased sucrose preference in sucrose preference test. In addition, we also found that HCE inhibited interleukin-1 beta (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) expression, as well as IDO activity in frontal cortex and hippocampus, which were increased in rats exposed to CUMS.

Conclusions

Combining with our previous studies, our present finding suggests that the anti-inflammatory property of HCE might play a crucial role in its antidepressant-like effect through, at least in part, the restoration or improvement of monoaminergic and neurotrophin systems.     

Propolis effect on Th1/Th2 cytokines production by acutely stressed mice

Aim of the study

Propolis has gained special attention due to its biological properties, however, little is known about its immunomodulatory effects in stress conditions. The purpose of this study was to investigate propolis effect on Th1/Th2 cytokines production by spleen cells of acutely stressed mice. Serum corticosterone concentration was determined as a stress indicator.

Materials and methods

Male BALB/c mice were submitted to restraint stress and treated with propolis (200 mg/kg) for 3 days. Supernatants of splenocytes cultures were assessed for Th1/Th2 cytokines determination.

Results

Regarding Th1 cytokines production, no alterations were seen in IL-2 production; however, IFN-γ production was inhibited in stressed mice, even when treated with propolis. As to Th2 cytokines, IL-4 was inhibited in stressed mice, but normal levels were seen when these animals were treated with propolis. No significant differences were found in IL-10 production between the experimental groups. Stressed groups (treated or not with propolis) showed higher corticosterone concentrations in comparison to control group.

Conclusions

Data suggest that propolis treatment was not able to counteract the stress-induced immunosuppressive effect on IFN-γ production; however, propolis showed an immunorestorative role, increasing IL-4 production in stressed mice, favoring humoral immune response during stress. Since the exact mechanisms of this natural product on immune system are still unclear, further studies are still required for a better comprehension of propolis use as a therapeutic alternative against the stress-induced negative effects that could lead to the development of various diseases.     

Therapeutic effects of extracts from Radix Toddaliae Asiaticae on collagen-induced arthritis in Balb/c mice

Aim of the study

Radix Toddaliae Asiaticae (RTA), also named “Sanbaibang”, is the dry root bark of Toddalia asiatica (L.) Lam. and has long been used as a traditional ethnic Chinese medicine for its considerable activity to alleviate pain and inflammation for patients suffering from rheumatism. It contains coumarin, alkaloids, triterpenes and volatile oils. Information regarding the anti-arthritis activity of RTA in vivo or in vitro is limited yet. In the present study, the aim is to investigate the therapeutic potential and underlying mechanisms of the ethyl alcohol extract (EtOH) and ethyl acetate fraction (EtOAc) from RTA on collagen II-induced arthritis (CIA) in mice.

Materials and methods

CIA animal model was performed by subcutaneous injection of type II bovine collagen (CII) on the 1st day and the 14th day of the experiment. Ethyl alcohol extract (542.8, 271.4, 135.7 mg/kg), ethyl acetate fraction (260.8, 130.4, 65.2 mg/kg) was orally administrated from the second antigen immunization for 3 weeks. Progression of edema of paws and knee joints was measured using a vernier caliper every 3 days from the 10th day after the first injection to the end of the experiment. The spleen index was measured and the knee joint changes were observed by pathological sections. ELISA was used to measure cytokines including tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1β), interleukin-6 (IL-6) and interleukin-10 (IL-10) in mice serum according to the manufacturer’s instructions.

Results

Administration of ethyl alcohol extract and ethyl acetate fraction remarkably reduced paws and joints swelling and decreased the spleen indexes. Histopathological examination demonstrated that RTA effectively protected bone and cartilage of knee joint from erosion, lesion and deformation versus those from the control group. Besides, the concentration of cytokines like TNF-α, IL-1β, IL-6 were significantly lower than the ones from the control group respectively, while cytokine like IL-10 was remarkably higher compare with the control group.

Conclusion

In this present study, it is demonstrated that administration of RTA has potential and therapeutic effect on CIA. The data suggests that RTA could have a contributory ethno-pharmacological role in improved management of RA patients.