Therapeutic effects of total alkaloids of Tripterygium wilfordii Hook f. on collagen-induced arthritis in rats

Ethnopharmacological relevance

Tripterygium wilfordii Hook f. is one of Traditional Chinese Medicines which is commonly used to treat rheumatoid arthritis (RA). The total alkaloids were the main constituent part of Tripterygium wilfordii Hook f. It has a great significance to study the effects of the total alkaloids of Tripterygium wilfordii Hook f. (ATW) on RA.

Aim of the study

This paper aims at investigating the therapeutic effect of ATW on RA and its possible mechanism, and providing a theoretical and experimental basis for the clinical use of ATW.

Materials and methods

The model of wistar rats of type II collagen-induced arthritis (CIA) was made, and the rats were perfused a stomach with ATW for 4 weeks continuously. Then the levels of interleukin (IL)-6, IL-8, tumor necrosis factor (TNF)-<alpha>, in the serum of CIA rats were detected by enzyme linked immunosorbent assay (ELISA), and the joint pathological section of CIA rats was observed by hematoxylin and eosin (HE) staining method and the expression of IL-6, IL-8, nuclear factor kappa B (NF-κB), TNF-α were measure by immunohistochemistry staining method.

Results

Compared with model group, ATW could significantly reduce paw swelling and suppresse articular cartilage degenerated. The results also found that there was significant reduction levels of IL-6, IL-8 and TNF-α in serum of CIA rats treated with ATW and ATW inhibited the expression of IL-6, IL-8, NF-κB, TNF-α in synovial tissue.

Conclusion

ATW not only could inhibit the symptom of CIA rats significantly but also could inhibit the production of IL-6, IL-8, TNF-α in serum and the expression of IL-6, IL-8, NF-κB and TNF-α in synovial tissue targeting the inflammatory. ATW would be a drug as a novel botanical drug for the treatment of RA.     

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.     

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.     

Protective effects of Huang-Lian-Jie-Du-Tang and its component group on collagen-induced arthritis in rats

Ethnopharmacological relevance

Huang-Lian-Jie-Du-Tang is a famous Traditional Chinese medicine consisting of Rhizoma coptidis (Coptis chinensis Franch, Ranunculaceae), Radix scutellariae (Scutellaria baicalensis Georgi, Labiatae), Cortex phellodendri (Phellodendron amurense Rupr. Rutaceae) and Fructus gardeniae (Gardenia jasminoide Ellis, Rubiaceae) in a weight ratio of 3:2:2:3.This formula was described by Wang Tao (in the Chinese Tang Dynasty) in his treatise “Wai Tai Mi Yao”. It has been used to treat inflammation, hypertension, gastrointestinal disorders, and liver and cerebrovascular diseases in the clinical practice of Traditional Chinese medicine, especially in treating inflammation for nearly two thousand years. However, the essential compounds in it have not been identified, and the mechanisms remain to be addressed. To investigate the protective effects of HLJDT and its component group (HLJDT–CG) on collagen-induced arthritis in rats.

Materials and methods

CIA was established in male Wistar rats with subcutaneous injection of type II bovine collagen at the base of the tail of animals. CIA rats were treated daily with oral administration of HLJDT aqueous extracts (270 mg/kg) or HLJDT–CG (40 mg/kg) once per day from day 6 to day 28. Rats in normal and vehicle control groups were given an equal volume of vehicle (0.9% saline) and 0.025 mg/kg Dexamethasone was given to the Standard group at the same time. The protective effect of them were assessed by measuring arthritis index, swelling, the cytokines such as TNF-α, IFN-γ and IL-17 in serum, type II collagen antibodies, splenocyte proliferation and so on.

Results

The results demonstrated that treatment of CIA rat with either HLJDT aqueous extracts or HLJDT–CG not only ameliorated the symptoms of arthritis, prevented joint damage but also reduced the serum levels of TNF-α, IFN-γ and IL-17 in CIA rats. Anti-CII antibodies showed the similar trend except that of IgG1. Furthermore, HLJDT aqueous extracts and HLJDT–CG administration also suppressed CII-induced proliferative response of splenocytes. More importantly, HLJDT–CG exhibited similar pharmacological activities as HLJDT aqueous extracts in all aforementioned experiments.

Conclusions

HLJDT aqueous extracts and HLJDT–CG could effectively ameliorate CII-induced arthritis and significantly suppress the immune response against CII with similar pharmacological efficacy. These findings suggest that HLJDT has therapeutic potential in RA treatment and HLJDT–CG can represent the effective-composite of HLJDT.     

Rapid-resolution liquid chromatography TOF-MS for urine metabolomic analysis of collagen-induced arthritis in rats and its applications

Aim of the study

Rheumatoid arthritis (RA) is a kind of autoimmune diseases characterized by persistent synovitis, systemic inflammation and autoantibodies. Huang-Lian-Jie-Du-Tang (HLJDT) is a traditional Chinese medicine (TCM) with anti-inflammatory activity. In the present study, a rapid-resolution liquid chromatography tandem time-of-flight mass spectrometry (RRLC-TOF-MS) based metabolomic study was developed to obtain a systematic view of the progression of RA and assess the efficacy of HLJDT and its components in collagen-induced arthritis (CIA) rats.

Materials and methods

Forty male Wistar rats were randomly divided into five groups, including model group, normal control group, dexamethasone group, HLJDT group and the mixture of 13 components of HLJDT group after immunized with bovine type II collagen. Urine samples for metabolomic study were collected on 8, 15, 22 day during the animal experiment.

Results

The pharmacological changes (swelling paws and arthritis scores) showed that prophylactic treatment with HLJDT and its components significantly suppressed the swelling of rats’ paws. By combining with partial least squares discriminant analysis, 24 potential biomarkers were identified and primarily involved in 12 metabolism pathways, such as tricarboxylic acids cycle metabolism, lipid metabolism, tryptophan metabolism and phenylalanine metabolism, which revealed a new insight into the RA network in vivo. These potential metabolites identified in CIA model need to be further investigated to prove their diagnostic and/or prognostic values for RA. Taking potential biomarkers found in the study as screening indexes, it revealed that HLJDT and its components could reverse the pathological process of RA through partly regulating the disturbed metabolic pathways.

Conclusions

This study indicated that the metabolomic strategy based on RRLC-TOF-MS is a useful tool to search potential biomarkers related to RA and to dissect the underlying mechanisms of TCM in treating RA.     

Influence of polysaccharide fractions isolated from Caltha palustris L. on the cellular immune response in collagen-induced arthritis (CIA) in mice. A comparison with methotrexate

Ethnopharmacological relevance

The extracts from Caltha palustris have been used in traditional Canadian and Asian medicine to treat arthritis and rheumatism.

Aims

The aim of the study was to investigate the anti-arthritis and immunomodulatory activity of the polysaccharide fractions B and C of Caltha palustris L. herbal extracts in collagen-induced arthritis (CIA) mice, an animal model of rheumathoid arthritis. The results were compared with those of methotrexate (MTX) treatment.

Materials and methods

CIA was induced in male and female DBA/1J mice by intradermal injection of chicken type II collagen in Freund’s complete adjuvant (cFA). Booster injection of collagen (in incomplete Freund’s adjuvant) was given on day 21 of the experiment. Mice were treated daily for 21 consecutive days with investigated fractions B or C at a dose of 10 mg/kg (the first dose was given 24 h after the booster) or phosphate buffered saline (PBS) (negative and positive control group). MTX was administered in parallel, intraperitoneally at three weekly cycles—every 48 h for 3 weeks at a dose of 6.6 mg/kg, the first dose was given on day 22 of the experiment. The severity of arthritis was evaluated by arthritic scores. Flow cytometry was used to investigate subsets of T lymphocytes in the thymus, and T and B lymphocytes in the spleen, and in mesenteric lymph nodes. T regulatory lymphocytes in the spleen were also quantified by means of flow cytometry. The levels of IL-2, IL-6, IL-10, IFN-γ and TNF-α in serum were also measured.

Results

The results revealed that fraction B significantly reduced the severity of joint swelling and erythema to a similar degree as MTX. It was also found that B fraction and MTX inhibited leucocytosis in peripheral blood caused by CIA, however the inhibitory effect of MTX persisted longer than that of fraction B. The analysis of lymphocyte T subsets demonstrated that both investigated fractions and MTX caused a partial or complete normalization in the percentage and the absolute number of CD4⁻CD8⁻ thymocytes (immature, double-negative cells), and increased the percentage of CD8⁺ T cells in peripheral lymphoid organs of mice with CIA. Moreover, an increase in the percentage of CD4⁺ thymic cells was observed after treatment with fraction B or MTX. Fraction C showed the weakest effect in normalization of the percentage and the absolute number of CD4⁻CD8⁻ thymus lymphocytes in mice with CIA. The potency of fraction B was comparable to MTX. A significant decrease in the percentage and the absolute count of splenic T-regulatory cells (CD4⁺CD25⁺FOXP3⁺) was observed after treatment with both Caltha palustris fractions. The inhibiting influence of investigated fractions on TNF-α serum concentration was significant and lasted longer in the case of fraction C. Production of other cytokines was modulated slightly (increase in IFN-γ) or markedly (decrease in IL-2).

Conclusion

The results of the experiment suggested that the administration of polysaccharide B fraction from Caltha palustris extract significantly suppressed the progression of CIA. These results are similar to those obtained in the case of MTX treatment. This indicates that fraction B may be a potent candidate for botanical anti-arthritic agent.     

The protective and healing effects of Si Shen Wan in trinitrobenzene sulphonic acid-induced colitis

Ethnopharmacological relevance

Si Shen Wan is a traditional Chinese herbal medicine formula for the treatment of diseases with diarrhea, such as ulcerative colitis, allergic colitis and chronic colitis. To investigate the protective and healing effects of Si Shen Wan in the experimental colitis induced by trinitrobenzene sulphonic acid, and to furture explore its mechanism of action.

Materials and methods

Rats with colitis treated with Si Shen Wan for 10 days. Colon wet weight, colon organ coefficient, colonic damage score and pathological change after trinitrobenzene sulphonic acid challenge were determined. The levels of MPO, MDA, GSH-PX, SOD and the expression of IL-4 and IL-10 mRNA in the colon were also measured.

Results

After treatment, colon wet weight, colon organ coefficient and colonic damage score were lower than that in the control group (p<0.05). MDA and MPO concentrations in the inflamed colonic tissues were decreased remarkably in the treated groups compared with that in the control group (p<0.05). But SOD level, IL-4 and IL-10 mRNA expression in the inflamed colonic tissues were obviously increased.

Conclusions

It is a potential path that protective effect of Si Shen Wan on impaired colonic mucosa rats with experimental colitis was accomplished by down-regulating the level of MDA and MPO, and up-regulating the level of SOD and the IL-4 and IL-10 mRNA expression in the colon mucosa.     

Identification of active compounds from Caesalpinia sappan L. extracts suppressing IL-6 production in RAW 264.7 cells by PLS

Ethnopharmacological relevance

Caesalpinia sappan L. is distributed in Southeast Asia and also used as herbal medicine for the treatment of various diseases such as burning sensations, leprosy, dysentery, osteoarthritis and rheumatoid arthritis (RA). The overproduction of IL-6 plays an important role in the prognosis of RA, but the active compounds from the extracts of Caesalpinia sappan L. suppressing IL-6 production remain unknown.

Aims of the study

Identifying the main active compounds of Caesalpinia sappan L. extracts inhibiting the IL-6 production in LPS-stimulated RAW 264.7 cells by partial least squares (PLS).

Materials and methods

Sixty-four samples with different proportions of compounds were prepared from Caesalpinia sappan L. by supercritical CO₂ fluid extraction (SCFE) and refluxing. Each of 64 samples was applied to RAW 264.7 cells with LPS to evaluate whether IL-6 production by LPS is affected by addition of each sample. The IL-6 production in medium was determined by ELISA and the inhibitory activity of each sample was analyzed. In addition, the fingerprints of these 64 samples were also established by ultra-performance liquid chromatography electrospray ionization tandem mass spectrometry (UPLC–MS). We used the PLS, a simplified method, to evaluate the results from IL-6 production and fingerprints.

Results

Each of 64 samples markedly suppressed LPS-induced IL-6 production in RAW cells. The fingerprints by UPLC–MS clearly revealed variations among 64 samples produced in different extract conditions. The PLS analysis with IL-6 production and fingerprints by UPLC–MS suggested that the peaks 71, 93, 150, 157, 168 have more influence on the inhibitory activity of Caesalpinia sappan L. extracts. The peaks 71, 93, 150 are likely representing sappanone A, protosappanin E and neoprotosappanin, respectively. The peaks 157 and 168 are still at large.

Conclusion

This is the first report that sappanone A, protosappanin E, neoprotosappanin and two unidentified compounds can be considered as possible active compounds that might inhibit IL-6 production. Further studies are needed to confirm the effectiveness of these five compounds on IL-6 production and possible mechanism.     

Anti-inflammatory and anti-osteoarthritis effects of fermented Achyranthes japonica Nakai

Ethnopharmacological relevance

Achyranthyes japonica Nakai (AJN) has been traditionally used to control pain and improve dysfunction in osteoarthritis (OA) patients.

Aim of the study

The objectives of the present study were to investigate anti-inflammatory and anti-osteoarthritis activities of fermented AJN (FAJN).

Materials and methods

Anti-inflammatory activity of non-fermented AJN (NFAJN) and FAJN was evaluated by in vitro assay using LPS-induced RAW 264.7 cells. In addition, their cartilage protective effects were also determined in vitro assay using SW1353 cell and in vivo model system using collagenase-induced arthritis (CIA) in rabbits. Moreover, we isolated and identified 20-hydroxyecdysone (20-HES) as a marker component in FAJN.

Result

FAJN showed stronger anti-inflammatory activity than NFAJN through inhibiting production of NO and PGE2 in LPS-induced RAW 264.7, and lowering levels of MMP-3 release in SW1353 cells treated with TNF-a. FAJN contained higher levels of 20-HES, as a marker component, than AJN. FAJN ameliorates the progress of OA by inhibiting local inflammation. It does this by regulating levels of TNF-a and IL-4, and protecting articular cartilage by preventing destruction of proteoglycan, collagens, and also preventing injury to chondrocytes.

Conclusion

Therefore, FAJN is a potential therapeutic agent for reduction of cartilage damage that occurs in OA.     

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.     

BAFF/BAFF-R involved in antibodies production of rats with collagen-induced arthritis via PI3K-Akt-mTOR signaling and the regulation of paeoniflorin

Ethnopharmacological relevance

Paeoniflorin (Pae) is extracted from the root of paeonia lactiflora which have attracted attention for anti-rheumatic and immune modulating properties.

Aim of the study

To investigate the role of PI3K/Akt/mTOR signaling mediated by BAFF/BAFF-R in antibodies production and the regulation of Pae on the signaling pathway in rats with collagen-induced arthritis (CIA).

Materials and methods

CIA rats were randomly separated into different groups and treated with Pae (25, 100 mg/kg) from day 18 to day 38 after immunization. The effects of Pae on B lymphocytes of CIA rats were evaluated by the levels of BAFF, anti-CII antibody, IgA, IgG and IgM, and the expressions of BAFF-R, PI3K, p-Akt and mTOR.

Results

In CIA rats, the levels of anti-CII antibody, IgA, IgG and IgM in serum enhanced, BAFF, BAFF-R, PI3K, p-Akt and mTOR were highly expressed. Pae (100 mg/kg) obviously decreased arthritis score, relieved ankle and paw swelling, improved spleen histopathology in CIA rats, decreased the levels of IgA, IgM, IgG and anti-CII antibody, and significantly decreased the expressions of BAFF, BAFF-R, PI3K, p-Akt and mTOR.

Conclusion

PI3K/Akt/mTOR signaling mediated by BAFF/BAFF-R participates in antibodies production by B lymphocytes of CIA rats. Pae had therapeutic effects on rats with CIA. These effects might be relative to regulating PI3K/Akt/mTOR signal mediated by BAFF/BAFF-R, and down regulate the antibodies production further.     

Clematichinenoside AR induces immunosuppression involving Treg cells in Peyer׳s patches of rats with adjuvant induced arthritis

Ethnopharmacological relevance

Clematichinenoside AR (AR) has been defined as a major active ingredient of triterpenoid saponins extracted from Clematidis Radix et Rhizoma, which is a traditional Chinese herbal medicine that has long been used in the treatment of rheumatoid arthritis (RA). To further explore the mechanism of AR in the treatment of RA, we investigated whether its immunomodulatory effects are related to Treg-mediated suppression derived from Peyer?s patches (PPs) in adjuvant induced arthritis (AIA) rat model.

Materials and methods

AR (8, 16, 32 mg/kg) was orally administered daily from Day 18 to Day 31 after immunization. The effect of AR on AIA rats was evaluated by hind paw swelling and histopathological examination. Percentages of CD4⁺CD25⁺Foxp3⁺ T regulatory cells were determined by flow cytometry. Levels of IL-10, TGF-β₁, IL-17A and TNF-α were measured by ELISA. Expressions of Foxp3 and RORγ in synovium were detected using immunohistochemical analysis.

Results

AR treatment significantly reduced paw swelling of AIA rats, and histopathological analysis confirmed it could suppress severity of established arthritis. AR treatment upregulated the percentages of CD4⁺CD25⁺Foxp3⁺ Treg cells among CD4+ T cells in PPs lymphocytes, and increased the levels of IL-10 and TGF-β₁ secreted from ConA-activated PPs lymphocytes, whereas decreased the levels of IL-17 A and TNF-α. Similar tendency of circulating CD4⁺CD25⁺Foxp3⁺ Treg cells percentages and serum cytokine levels were observed. Moreover, AR decreased the expression levels of Foxp3 and RORγ in joint synovial membrane.

Conclusions

In conclusion, these results suggested AR has a potent protective effect on the progression of AIA, probably by augmenting CD4⁺CD25⁺Foxp3⁺ Treg cells in PPs to induce immunosuppression, and modulating the balance between Treg cells and Th17 cells systemically. These findings may help to develop AR as a potent immunosuppressive agent for the treatment of RA.     

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.     

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.     

Extracts of Arisaema rhizomatum C.E.C. Fischer attenuate inflammatory response on collagen-induced arthritis in BALB/c mice

Aim of the study

Arisaema rhizomatum C.E.C. Fischer (ARCF), called as “Xuelijian”, a local herb just growing in China, has been used as a traditional ethnic Chinese medicine for long because of its remarkable activity to alleviate pain and inflammation for patients suffering from rheumatism among the people with weak side-effect. However, rare study on the anti-arthritic activity of ARCF has been reported in vivo. The aim of this study is to investigate the protective effect of the herb on collagen-induced arthritis in mice and explore the potential immunological mechanisms.

Materials and methods

CIA was induced in male BALB/c mice by been subcutaneously injected type II bovine collagen (CII) for twice. The combined MeOH extract (ME) of ARCF rhizome was successively partitioned into four fractions with petroleum ether (PE), ethyl acetate (EE), n-butyl alcohol (n-BE) and water (WE). After the second collagen immunization, mice were administered orally with different doses of ME, EE and n-BE (ME 130, 261, 522 mg kg⁻¹; EE 10.2, 20.4, 40.8 mg kg⁻¹; n-BE 52, 104, 208 mg kg⁻¹) every other day for 3 weeks. The progression of edema of paws and knee joints was inspected by using a vernier calliper 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 destruction was observed by pathological sections. Levels of inflammatory cytokines including tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1β), interleukin-6 (IL-6), interleukin-33 (IL-33 or IL-1F11) and rheumatoid factor (RF) in serum were measured by ELISA.

Results

Administration of ME, EE and n-BE significantly suppressed paws and joints swelling and reduced the spleen indexes. Pathological examination demonstrated that ARCF effectively protected anklebone and cartilage from being eroded versus vehicle-treated mice. Moreover, the serum levels of inflammatory cytokines TNF-α, IL-1β, IL-6, IL-33 and RF were markedly lowered in ARCF treated groups compared with the control group (p < 0.05).

Conclusion

Our studies demonstrate that administration of ARCF is obviously suppressed the progression of CIA. The anti-arthritic effectiveness of ARCF will make the herb a strong candidate for further clinical trials on RA patients.     

Therapeutic effects of total coumarins from Urtica dentata Hand on collagen-induced arthritis in Balb/c mice

Ethnopharmacological relevance

Urtica dentata Hand (UDH), the root of Laportea bulbifera (Sieb. et. Zucc.) Wedd, has long been utilized in traditional Chinese medicine for the treatment of rheumatoid arthritis and some other autoimmune diseases. Coumarins are the main active principles contributing to UDH's efficacy, but the mechanisms have not been fully clarified.

Aim of study

To explore effects of total coumarins (TC) isolated from UDH on the development of type II collagen (CII)-induced arthritis (CIA) in Balb/c mice.

Materials and methods

Arthritis was induced in Balb/c mice by immunization with an emulsion of 200 mg CII and complete Freund's adjuvant (CFA). The CIA mice were then given with a suspension of TC or saline by intragastric (i.g.) administration every other day. The incidence and severity of disease and histopathology of inflammation were assessed. Inflammatory response was determined by measuring the levels of different inflammation mediators in serum. The effect of TC on differentiation of CD4⁺CD25⁺ Foxp3⁺Treg cells was examined by flow cytometry. The phenotype of bone marrow-derived dendritic cells (DCs), T-bet mRNA level and IL-12p70 secretion by DCs were also detected.

Results

Pharmacologically, treatment with TC for type II collagen induced arthritis in mice through oral administration displayed significant and dose-dependent drop of clinical arthritis score and paw swelling, compared with the untreated CIA mice. Pathologic changes showed that TC protected tissues against bone destruction, whereas an almost complete destruction occurred in the CIA model group. The protective status was associated with a substantial decrease in the production of IFN-γ and IL-2, an increase of IL-10 and TGF-β and suppressive expression of T-bet in DCs. TC also induced the generation of CD4⁺CD25⁺ Treg cells with a Treg phenotype Foxp3. TC-treated DCs were characterized as low expression of MHC class II and CD86 molecules, as well as a reduction of IL-12p70.

Conclusions

Our data suggest that TC provides substantial therapeutic protection against CIA by eliciting immune tolerance and it would be a valuable candidate for further investigation as a new anti-arthritic agent.