Stimulative effects of Ulmus davidiana Planch (Ulmaceae) on osteoblastic MC3T3-E1 cells

Ulmus davidiana Planch (Ulmaceae) has long been known to have anti-inflammatory and protective effects on damaged tissue, inflammation and bone among other functions. To treat rheumatoid arthritis (RA), a herbal medicine, Ulmus davidiana Planch (Ulmaceae) extract (UD) is being used in traditional oriental medicine. The effect of UD on the proliferation and osteoblastic differentiation in non-transformed osteoblastic cells (MC3T3-E1) was studied. UD dose-dependently increased DNA synthesis (significant at 5-20 microg/ml). UD increased alkaline phosphatase (ALP) activity and prolyl hydroxylase activity of MC3T3-E1 cells (5-20 microg/ml). Antiestrogen tamoxifen eliminated the stimulation of proliferation and ALP activity of MC3T3-E1, which was induced by UD. UD at concentrations ranged from 30 to 100 microg/ml inhibited prostaglandin E2 production in MC3T3-E1. These results indicate that UD directly stimulates cell proliferation and differentiation of osteoblasts. These results also suggest and UD is effective for bone anti-resorptive action in bone cells.     

Suppressive effects of Chelidonium majus methanol extract in knee joint, regional lymph nodes, and spleen on collagen-induced arthritis in mice

Chelidonium majus L. has multiple applications in Korean traditional medicine because of its anti-tumoral, cytotoxic, anti-inflammatory and anti-microbial activities and has long been known to have anti-inflammatory effects. However, no study on the anti-arthritic activity of Chelidonium majus has been reported in vivo. Rheumatoid arthritis (RA) is a systemic autoimmune disease with chronic inflammation characterized by hyperplasia of synovial cells in affected joints, which ultimately leads to the destruction of cartilage and bone. Cytokine production and gene expression were assessed during CIA (collagen-induced arthritis) model mice in knee joint, lymph node (LN), and spleen, using ELISA and competitive RT-PCR. DBA/1J mice were immunized with bovine type II collagen. After a second collagen immunization, mice were treated with CME orally at 400, 40mg/kg once a day for 4 weeks. The severity of arthritis within the knee joints was evaluated by histological assessment of cartilage destruction and pannus formation. Administration of CME significantly suppressed the progression of CIA and inhibited the production of TNF-alpha and IL-6 in spleen and lymph node. The erosion of cartilage was dramatically reduced in mouse knees after treatment with CME. In conclusion, our results demonstrates that CME significantly suppressed the progression of CIA and that this action was characterized by the decreased production of TNF-alpha, IL-6, IFN-gamma, B cells, gammadelta T cells (in spleen) and increased proportion of CD4+CD25+ regulatory T cells in vivo. In the serum of CME-treated mice, the levels of IgG and IgM RA factor were decreased.     

Anti-inflammatory effect of bee venom on type II collagen-induced arthritis

Bee venom (BV) has been used to relieve pain and reduce inflammation in traditional Oriental medicine, especially in chronic inflammatory diseases such as rheumatoid arthritis (RA). We previously reported that the BV injection into a traditional acupuncture point (Zusanli) reduced arthritis-associated edema and nociceptive responses in Freund's adjuvant-induced arthritis in rats (Kwon et al., 2001). This study was designed to evaluate the anti-inflammatory and anti-cytokine effect of BV on a murine type-II collagen-induced arthritis (CIA) model. Male mice were immunized by spontaneous injection of 100 microg of an emulsion of bovine type-II collagen and complete Freund's adjuvant (CFA), with a booster injection after 2 weeks. In the experimental group, 0.1 ml BV was injected at acupuncture point (Zusanli) near both knees twice a week for a total of 5 times. In the control group, normal saline was injected at the same frequencies. These injections began 5 weeks after the first collagen injection. Starting the 3rd week after the first collagen injection, we examined limb swelling and severity of arthritis twice a week. At 8 weeks, mice were sacrificed and synovial tissue was examined with the light microscope and serum cytokines (IL-1beta and TNF-alpha) were measured by ELISA. The incidence of arthritis, the mean arthritis index and the number of arthritic limbs were significantly lower in the treatment compared to the control group (63% versus 75%, 3.4% versus 8.5%, 23% versus 75%, respectively). Among the serum proinflammatory cytokines, the production of TNF-alpha in the BV group was suppressed compared to the control group (59 +/- 4.5 versus 99.5 +/- 6.5, p < 0.05), but IL-1beta was not suppressed. The examination of the histopathology of the joints of murine CIA showed decreased inflammation signs and less lymphocyte infiltration after BV acupuncture therapy. Acupuncture therapy with BV suppressed the development of arthritis and caused inhibition of the immune responses in type-II collagen-induced arthritis.     

Inhibition of Ulmus davidiana Planch (Ulmaceae) on bone resorption mediated by processing of cathepsin K in cultured mouse osteoclasts

Ulmus davidiana Planch (Ulmaceae) (UD) has long been known to be antiinflammatory in traditional Korean medicine. This experiment investigated the effects of UD on bone resorption using bone cell culture. Different concentrations of crude extract of UD were added to mouse bone cell culture. The mitochondrial activity of the bone cells after exposure of UD was determined by colorimetric 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT). It was demonstrated that UD has potential effects on bone cell culture without cytotoxicity. The most effective concentration of UD in bone cells was 100 microg/mL. Cathepsin K (Cat K) is the major cysteine protease expressed in osteoclasts and is thought to play a key role in matrix degradation during bone resorption. When mouse long bone cells including osteoclasts and osteoblasts were treated with UD, UD prevented the osteoclast-mediated intracellular processing of Cat K, suggesting that UD may disrupt the intracellular transport of pro Cat K. Since secreted proenzymes have the potential to reenter the cell via the mannose-6-phosphate (M6P) receptor, to prevent this possibility, UD was tested in the absence or presence of M6P. Inhibition of Cat K processing by UD was observed in a dose-dependent manner. Furthermore, the addition of M6P resulted in enhanced potency of UD. UD dose-dependently inhibited in vitro bone resorption with a potency similar to that observed for inhibition of Cat K processing.     

Preliminary study of effects and mechanisms of Qingpeng paste on type II collagen-induced arthritis in rats

OBJECTIVE: To investigate the effect of Qingpeng paste (QP) on collagen-induced arthritis (CIA) in rats. METHOD: CIA was established in female Wistar rats with injection of type II bovine collagen at the base of the tail of animals. CIA rats were treated daily with external administration of different doses of QP or voltaren beginning on the day after the onset of arthritis (day 1) until day 20. Paw swelling rate and the serum levels of IL-1 beta were determined. Moreover, the expression of TNF-alpha and IL-alpha and histopathological changes in the arthritic joints were also observed. RESULT: QP markedly suppressed the paw swelling rate of arthritic rat, reduced the expression of TNF-alpha and IL-alpha in synovial membrane. Histopathological changes in the arthritic joints were also significantly ameliorated in the QP-treated versus vehicle-treated rats. However, the elevated serum levels of IL-1 beta in arthritic rats were not influenced by QP. CONCLUSION: The present findings demonstrate the protective property of QP on collagen-induced arthritis, mechanisms underlying it may be related to reduce the expression of IL-1alpha and TNF-alpha in synovial membrane.     

Suppression of the onset and progression of collagen-induced arthritis in rats by QFGJS, a preparation from an anti-arthritic Chinese herbal formula

QFGJS is an herbal preparation, and its pronounced effectiveness in treating adjuvant-induced arthritis (AIA) has been previously demonstrated. We herein aimed to confirm its anti-arthritic effect on collagen-induced arthritis (CIA) in rats. CIA was established in female Wistar rats with intradermal injection of type II bovine collagen at the base of the tail of animals. CIA rats were treated daily with oral administration of different doses of QFGJS beginning on the day of the induction of arthritis (day 0, the prophylactic treatment) or on the day after the onset of arthritis (day 13, the therapeutic treatment) until day 30. The results showed that prophylactic treatment with QFGJS significantly suppressed the onset of arthritis, and therapeutic treatment with QFGJS markedly reduced paw swelling and ESR levels even in the established CIA. Radiologic and histopathologic changes in the arthritic joints were also significantly reduced in the QFGJS-treated versus vehicle-treated rats. Moreover, the serum levels of pro-inflammatory cytokines TNF-alpha, IL-1beta, and IL-6 were markedly lowered in the QFGJS-treated rats. Hence, our studies demonstrate the quality, safety, and effectiveness of QFGJS as an anti-arthritic agent, which makes QFGJS a strong candidate for further clinical trials on rheumatoid arthritis (RA) patients.     

Anti-inflammatory and immune-modulating effect of Ulmus davidiana var. japonica Nakai extract on a macrophage cell line and immune cells in the mouse small intestine

Ethnopharmacological relevance

Traditional Oriental medicine has utilized the barks of the stem and root of Ulmus davidiana var. japonica Nakai (UD) to treat inflammatory disorders.

Aim of the study

The purpose of the present study was to evaluate UD's anti-inflammatory and immune-modulating effects on a lipopolysaccharide (LPS)-stimulated macrophage cell line and small-intestinal lamina propria (LP) cells, respectively.

Materials and methods

RAW 264.7 cells were stimulated with LPS in the presence of various concentrations of a UD water-soluble extract. Cell viability, nitric oxide (NO) production, and the level of inflammatory cytokines synthesis were measured. Among the mice receiving the UD water-soluble extract, changes in the LP cell populations and immunoglobulin (Ig)A production were evaluated.

Results

The UD water-soluble extract inhibited LPS-induced NO synthesis and inflammatory cytokine production in a RAW264.7 macrophage-like cell line. Small-intestinal LP cells isolated from mice that received the UD extract displayed a decrease in the side scatter of medium-to-high cells. Those LP cells isolated from the UD-treated mice also showed a marked decrease of intracellular IgA. However, UD administration had no apparent effect on the synthesis of systemic inflammatory cytokines.

Conclusions

These results suggest that UD water-soluble extracts have anti-inflammatory properties and, as such, can be used to promote intestinal immune-homeostatic conditions.     

Protective effects of Ulmus davidiana var. japonica against OVA-induced murine asthma model via upregulation of heme oxygenase-1

Aim of the study

Traditionally, the stem and root bark of Ulmus davidiana var. japonica (Ulmaceae) are Korean herbal medicines used for anti-inflammatory and anticancer therapy. In this study, we investigated the protective effects of Ulmus davidiana var. japonica ethanolic extract (UD) in a murine asthma model. Furthermore, we determined whether heme oxygenase (HO)-1 is required for the protective activity of UD.

Materials and methods

Airways of ovalbumin (OVA)-sensitized mice exposed to OVA challenge developed eosinophilia, mucus hypersecretion and increased cytokine levels. UD was applied 1 h prior to OVA challenge. Mice were administered UD orally at doses of 100 and 200 mg/kg once daily on days 18–23. Bronchoalveolar lavage fluid (BALF) was collected 48 h after the final OVA challenge. Levels of interleukin (IL)-4 and IL-5 in BALF were measured using enzyme-linked immunosorbent assays (ELISAs). Lung tissue sections 4 μm in thickness were stained with Mayer's hematoxylin and eosin for assessment of cell infiltration and mucus production with PAS (periodic acid shift reagent) staining, in conjunction with ELISA, immunohistochemistry and Western blot analyses for HO-1 protein expression.

Results and conclusion

Orally administered UD significantly inhibited the number of OVA-induced inflammatory cells and IgE production, along with reduced T-helper (Th)2 cytokine levels, such as IL-4 and IL-5, in BALF and lung tissue. In addition, UD induced a marked decrease in OVA-induced reactive oxygen species (ROS), inflammatory cell infiltration and mucus production in lung tissue. These effects were correlated with HO-1 mRNA and protein induction. Our results indicate that UD protects against OVA-induced airway inflammation, at least in part, via HO-1 upregulation.     

京尼平苷对类风湿性关节炎大鼠血清IL-1β和TNF-α的影响

目的:研究京尼平苷对类风湿性关节炎大鼠血清IL1β和TNFα的影响,进一步探讨该药物的作用机制。方法:建立Ⅱ型胶原蛋白诱导大鼠类风湿性关节炎模型。观察京尼平苷对实验性大鼠足肿胀的抑制作用;检测血清IL1β和TNFα的水平。结果:与模型组比较,京尼平苷可以延缓对侧肢体足肿胀发生的时间,抑制大鼠足肿胀的程度(P<0.01);用药后,中、高剂量给药组血清IL1β和TNFα水平明显低于模型组(P<0.01)。结论:京尼平苷在类风湿性关节炎大鼠中可以下调致炎因子IL1β和TNFα的水平,提示这可能与该药抑制实验性类风湿性关节炎病情进展密切相关。     

雷公藤甲素配伍甘草酸对CIA大鼠血清TNF-α,IL-10的影响

目的：观察雷公藤甲素与甘草酸配伍对CIA模型大鼠关节肿胀和血清TNF-α,IL-10水平的影响。方法：使用Ⅱ型胶原诱导出成功的关节炎模型大鼠60只,并随机分为模型组、雷公藤甲素组、甘草酸组、雷草1组、雷草2组和雷草3组,另设空白组。以关节炎指数和关节肿胀度作为疗效评判标准,并采用ELISA方法检测大鼠血清中抗Ⅱ型胶原抗体,TNF-α和IL-10的水平。结果：经雷公藤甲素配伍甘草酸治疗后,模型大鼠的关节炎指数和关节肿胀度均显著下降(P＜0.01),血清中抗Ⅱ型胶原抗体和TNF-α水平显著下降,而IL-10水平显著升高(P＜0.05,P＜0.01)。结论：甘草酸配伍雷公藤甲素治疗CIA,可以在保持疗效的前提下减少雷公藤甲素的用量。     

二仙除痹汤对大鼠胶原诱导型关节炎的影响

目的探讨二仙除痹汤对大鼠胶原诱导型关节炎（CIA）的作用及机制。方法取健康Wistar大鼠,随机分为正常组、模型组及二仙除痹汤低、高剂量组,除正常组外其余各组建立大鼠CIA模型。二仙除痹汤低、高剂量组于造模后第7日起分别给予二仙除痹汤8、16 g/kg灌胃,每日1次,连续4周。评估发病率及临床积分,观察关节组织病理形态和放射学改变,检测关节组织匀浆上清和血清中促炎细胞因子白细胞介素（IL）-1β、IL-6、肿瘤坏死因子（TNF）-α以及血清炎症介质前列腺素（PG）E2的含量变化。结果模型组大鼠CIA发病率100%,关节炎最高临床积分8.2,踝关节出现明显的炎症细胞浸润、血管翳形成、软骨和骨破坏等组织病理学变化以及关节间隙狭窄或增宽、关节面虫蚀样破坏等影像学改变,关节和血清中IL-1β、TNF-α、IL-6以及血清PGE2含量明显升高。与模型组相比,二仙除痹汤低、高剂量组可显著降低关节炎的发病率（分别为60%、40%）、临床积分（分别为5.7、3.8）和改善关节组织病理学改变,二仙除痹汤高剂量组还能明显抑制骨破坏程度,降低关节和血清中IL-1β、TNF-α、IL-6及PGE2的含量,二仙除痹汤低剂量组的亦显著抑制血清中IL-1β、IL-6以及关节中IL-6的含量。结论二仙除痹汤能控制大鼠CIA病情的发生发展,其机制可能与对系统和关节局部促炎细胞因子和炎症介质的有效抑制有关。     

阿克他利对Ⅱ型胶原性关节炎小鼠的治疗作用及部分机制研究

 目的 研究阿克他利(Acta)对小鼠Ⅱ型胶原性关节炎(CIA)的治疗作用。方法 采用Ⅱ型胶原(CⅡ)乳剂皮内注射诱导的小鼠CIA模型。在此基础上,检测小鼠足肿胀、免疫功能的改变,以及对CⅡ的迟发性变态反应(DTH)和血清中抗CⅡ抗体的测定,同时进行了病理组织学的检查。结果 Acta(10,30,90 mg·kg^-1)ig对CⅡ诱导的小鼠足肿胀有明显的抑制作用;体外研究发现,Acta(10,30,90 mg·kg^-1)能使CIA小鼠过高的ConA增殖反应和IL-2的产生恢复至接近正常,同时对CIA小鼠过高的IL-1产生有明显的抑制作用;Acta(10,30,90 mg·kg^-1)ig可以明显减轻CⅡ诱发迟发性变态反应(DTH),但Acta对CIA小鼠体内的抗体的产生无显著影响。病理学检查表明,Acta(10,30,90 mg·kg^-1)ig可以减轻CIA小鼠的骨膜增生和软骨破坏。结论 Acta对CIA小鼠具有治疗作用,该作用可能是通过细胞免疫调节实现的。     

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.     

Effects and mechanisms of total glucosides of paeony on synoviocytes activities in rat collagen-induced arthritis

The aim of the study was to investigate the effects of TGP, an active compound extracted from the roots of Paeonia lactiflora Pall, on the activities of synoviocytes in rats with collagen-induced arthritis (CIA) and its possible mechanisms. CIA was induced in male Sprague-Dawley (SD) rats immunized with chicken type II collagen (CII) in Freund's complete adjuvant (FCA). Synoviocytes proliferation was determined by 3-(4, 5-2dimethylthiazal-2yl) 2, 5-diphenyltetrazoliumbromide (MTT) assay. Tumor necrosis factor alpha (TNF-alpha), interleukin-1 (IL-1), prostaglandin E(2) (PGE(2)) and cyclic adenosine monophosphate (cAMP) levels in synoviocytes were measured by radioimmunoassay (RIA). E-prostanoid (EP)(2) and EP(4) receptors were analyzed by Western blot analysis. The results showed that TGP significantly inhibited the proliferation of synoviocytes, decreased the production of IL-1, TNF-alpha and PGE(2) and elevated the levels of cAMP. Further study showed that TGP could up-regulate the expression of EP(2) and EP(4). These results indicated that TGP might exert its anti-inflammatory effects through inhibiting the production of pro-inflammatory mediators in synoviocytes of CIA rats, which might be associated with its ability to regulate cAMP-dependent EP(2)/EP(4)-mediated pathway.     

桑桂通痹剂对胶原诱导关节炎小鼠病理形态学及血管内皮生长因子、基质金属蛋白酶3的影响

目的观察桑桂通痹剂对胶原诱导关节炎（CIA）模型小鼠踝关节血管翳数目、关节炎指数及血管内皮生长因子（VEGF）、基质金属蛋白酶3（MMP-3）的影响,探讨其作用靶点。方法将50只DBA／1J小鼠随机分为正常纽、模型组、桑桂通痹剂组、甲氨蝶呤组、雷公藤多甙片组,除正常组之外,其余各组使用II型胶原诱导小鼠造成CIA模型,桑桂通痹剂组、甲氨蝶呤组、雷公藤多甙片于造模后开始给药,共计28d。按评分法评价关节炎指数,病理观察小鼠踝关节血管翳数目,免疫组化法检测VEGF、MMP-3蛋白表达。结果桑桂通痹剂组可降低模型小鼠的关节炎指数;减少由滑膜细胞和毛细血管增生所形成的血管翳数目,VEGF、MMP-3表达也有所降低。结论桑桂通痹荆可能是通过抑制VEGF、MMP-3表达,减少模型小鼠的关节炎指数和血管翳数目,从而起到治疗作用。     

紫草素对胶原诱导关节炎的药效学研究

目的：研究紫草素对胶原诱导的关节炎（collageninducedarthritis,CIA）小鼠的疗效。方法：将紫草素高低两种剂量给CIA小鼠灌胃,连用30天,对小鼠一般状态进行观察,用关节炎评分评估CIA小鼠关节炎病情;小鼠足测量仪检测关节肿胀程度;组织学染色评价膝关节的软骨和骨破坏。结果：紫草素高剂量组与对照组相比,可以降低CIA小鼠的关节炎评分,阻止病理学进展;延缓CIA小鼠体重下降程度,降低后爪肿胀程度。结论：紫草素能够有效控制CIA小鼠的炎症进展,阻止骨质破坏,免疫抑制作用可能是其治疗胶原诱导的关节炎的机制。     

Effect of pine pollen extract on experimental chronic arthritis

The effects of pine pollen extract (PE) on Freund's complete adjuvant (FCA)‐induced arthritis and collagen‐induced arthritis (CIA) were investigated. The oral administration of PE (100 and 200 mg/kg per day) for 21 days after subcutaneous immunization with FCA, significantly reduced hindpaw swelling and the production of inflammatory cytokines (TNF‐α, IL‐1β and IL‐6) compared with the FCA‐induced arthritis group. Treatment with the PE (100 mg/kg) also decreased the serum levels of LDL‐cholesterol and increased HDL‐cholesterol contents compared with those of the arthritis group. Since CIA is a model of some types of human autoimmune rheumatoid arthritis (RA), the study examined whether PE is efficacious against CIA in mice and investigated the possible antioxidant potential of PE on CIA. Arthritis in DBA/1J mice was induced by subcutaneous immunization with bovine type II collagen. PE (100 and 200 mg/kg) was orally administered once daily for 49 days after initial immunization with type II collagen. The arthritis score and paw edema were markedly suppressed in the groups treated with PE. Moreover, administration of PE (100 mg/kg) for 49 days reduced the serum levels of rheumatoid factor, anti‐type II collagen antibody, TNF‐α, IL‐1β, IL‐6, protein carbonyl, advanced glycation endproducts, malondialdehyde and LDL‐cholesterol compared with that of CIA mice. These results suggest that the pine pollen might be beneficial in the treatment of chronic inflammatory disorders. Copyright © 2009 John Wiley & Sons, Ltd.