Differential effects of methotrexate and liposomally conjugated methotrexate in rat adjuvant-induced arthritis.

In this study we evaluated the comparative efficacy of free and liposomally conjugated methotrexate on both disease induction and suppression of acute inflammation in rat adjuvant-induced arthritis. Rats were given either empty liposomes (E-LIPO), free methotrexate (MTX) or the liposomally conjugated methotrexate (MTX-LIPO) at a dose of 100 micrograms/day for 7 consecutive days by the intravenous route. When MTX treatment was initiated on the day of arthritis induction the drug suppressed but did not abolish the development of joint inflammation. Free MTX had no significant anti-inflammatory effect upon an established arthritis when dosing was commenced on day 11 post-adjuvant induction. Conversely, MTX-LIPO did not affect the progression of the arthritis when dosing was started on day 0, but exerted a significant anti-inflammatory effect on an established arthritis. MTX-LIPO treatment was significantly less haematotoxic than free MTX.     

Effects of bisphosphonates on joint damage and bone loss in rat adjuvant-induced arthritis

Objective and design: Examination of the effects of bisphosphonates on joint damage and generalized bone loss.Materials: Adjuvant-arthritis was induced by injection of Mycobacterium butyricum into the footpad of the right hind paw of Lewis rats (8 animals/group) on day 0.Treatment: Arthritic rats were treated with the vehicle (saline), etidronate or alendronate (subcutaneously, daily 5 times a week for 3 weeks from day 1 to day 21). Experiment-1: Etidronate (0.1, 0.5, 2.5, 12.5 mg/kg) or alendronate (0.02, 0.1, 0.5, 2.5 mg/kg), Experiment-2: Etidronate (2.5, 5, 10mg/ kg) or alendronate (0.001, 0.01, 0.1 mg/kg).Methods: In the adjuvant-injected side of the hind limbs, paw swelling was evaluated at 1-week intervals, and bone mineral density (BMD) in the proximal tibia, histopathology and radiographical findings in the tibio-tarsal region were evaluated at the time of sacrifice (on day 21).Results: In all treatment schedules, both bisphosphonates significantly prevented paw swelling and bone loss. Alendronate reduced paw swelling at higher doses (over 0.1 mg/ kg) compared with its effect on BMD decrease (over 0.001 mg/kg). In contrast, etidronate reduced paw swelling and joint damage at doses similar to those (over 2.5 mg/kg) prevented BMD decrease.Conclusions: Both etidronate and alendronate are effective in reducing arthritic damage, but their effective dose ranges for inflammatory responses and BMD decrease clearly differ; i.e., the etidronate dose ranges for anti-inflammatory and anti-resorptive effects are similar, whereas the dose range for anti-inflammatory effects of alendronate is 100-fold higher than that for its anti-resorptive effects.Received 21 January 2003; returned for revision 14 July 2003; accepted by M. Katori 15 August 2003     

Inhibitory effect of allopurinol on adjuvant arthritis in rats

The effect of Allopurinol (ALLO), on adjuvant arthritis was studied in rats and compared with the effect of indomethacin (IND). Drugs were given by intraperitoneal injection for each day beginning from the day of adjuvant injection (day 0) and continued until the 16th day. Paw swelling was measured on days 4, 17 and 29, and secondary lesions were assessed on days 17 and 29. Polymorphonuclear leukocyte (PMNs) count was also evaluated on day 17.ALLO, at relatively high doses (25–50 mg/kg), reduced paw swelling of the adjuvant-injected extremity on day 4; lower doses (6.25–12.5 mg/kg), however, elicited the same inhibitory effect on day 17. IND (0.25 mg/kg) also prevented paw swelling on days 4 and 17. Both ALLO and IND reduced the secondary lesions on days 17 and 29 and prevented the increase in polymorphonuclear leukocytes during the development of adjuvant arthritis. Possible mechanisms of the antiinflammatory effect of ALLO in adjuvant arthritis are discussed.     

复方雷公藤凝胶剂对大鼠佐剂型关节炎的疗效及肝毒性研究

目的 探讨不同剂量复方雷公藤凝胶剂(TWCG)外用对佐剂型关节炎(AA)大鼠的治疗作用及其对肝功能的影响.方法 以弗氏完全佐剂(FCA)诱导大鼠关节炎模型,用不同剂量TWCG对其外敷治疗,以扶他林乳胶剂为阳性对照,以凝胶基质为空白对照.观察各组大鼠关节肿胀度;ELISA测定大鼠血清中肿瘤坏死因子-α(TNF-α)、白细胞介素-4(IL-4)的含量;免疫组织化学方法检测踝关节软骨中基质金属蛋白酶-3(MMP-3)的含量;以全自动生化分析仪检测大鼠血清中的谷丙转氨酶(ALT)、谷草转氨酶(AST)水平来观察其肝毒性.结果 与空白对照组相比,扶他林乳胶剂组及不同剂量TWCG组均能显著减轻大鼠关节肿胀度(P＜0.05);能明显降低血清中TNF-α的含量并增加血清中IL-4含量(P＜0.05),能显著降低关节组织中MMP-3的含量(P＜0.05),且中、高剂量TWCG组效果明显优于扶他林及小剂量TWCG组(P＜0.05);但各组大鼠之间ALT、AST差异无统计学意义(P＞0.05).结论 TWCG对AA大鼠有明显的抗炎消肿作用,能有效保护关节软骨组织,同时无明显的肝脏毒性.     

瑞香狼毒甲醇提取物抗瘤机理研究

目的 研究瑞香狼毒甲醇提取物的抗瘤机理。方法 用不同剂量的瑞香狼毒甲醇提取物(简称醇提物)，观察其体内对荷瘤鼠和所荷肿瘤生长的作用；MTT法研究其体内对荷瘤鼠脾细胞转化及NK活性，体外对正常鼠脾细胞生长、转化和NK活性以及对3株肿瘤细胞增殖的影响；流式细胞术和透射电镜检测其体外对K562的细胞周期和凋亡及p53、bcl-2和c-myc基因表达的影响；台盼蓝拒染法检测其体外对K562增殖曲线的影响。结果 体内，每天5μg/kg醇提物可抑制肿瘤生长，提高荷瘤鼠免疫功能。体外，0．1～l0μg/ml醇提物可刺激脾细胞增殖和协同最适量和亚适量ConA刺激脾细胞转化，0．1～10μg/ml醇提物可提高脾细胞NK杀伤活性，0．1～l0μg/ml醇提物对K562、S180和YAC-1的增殖均有抑制作用，K562最为敏感。0．5μg/ml醇提物作用的K562，处于G0/G1期的细胞和凋亡细胞显著增加，p53基因表达显著增加，bcl-2和c-myc基因表达显著下降。K562在0．5μg/ml醇提物作用24h后洗去或不洗去培养9d，细胞虽仍存活，但密度却不增加。结论 醇提物在一定剂量范围内可抑制肿瘤细胞生长，提高荷瘤鼠免疫功能。抗瘤机理与其可刺激脾细胞增殖，协同ConA刺激脾细胞转化和提高脾细胞NK杀伤活性及阻滞肿瘤细胞周期，抑制其分裂增殖，促进其凋亡有关。     

树突状细胞对佐剂性关节炎作用的实验研究

本文建立了大鼠佐剂性关节炎模型，经体内应用ＷＺＤ５（大鼠树突状细胞单抗）和输注新鲜分离的同系大鼠淋巴树突状细胞，以模型鼠的关节肿胀率和关节组织的病理改变、ＣｏｎＡ刺激淋巴细胞增殖反应和循环免疫复合物等为指标，来分析树突状细胞在佐剂性关节炎发病过程中的作用。结果表明：该细胞具有促进佐剂性关节炎的作用，而ＷＺＤ５则能减少关节肿胀率、减轻病变、缩短病程。     

乙醇对体外培养大鼠大脑皮层神经细胞分化的影响

目的:观察神经细胞微管相关蛋白2(microtubeasso ciated protein-2,MAP2)、孤儿核受体1(nuclear receptor-related factor 1,NURR1)、拓扑异构酶Ⅱβ(DNA topoisomereⅡβ,TopoⅡβ)在乙醇毒性作用下的表达变化,探讨MAP2、NURR1、TopoⅡβ与乙醇导致的神经细胞分化异常的相关关系。方法:取新生鼠(出生24 h内),75%酒精消毒,断头取脑,分离皮层神经细胞接种于培养板,给予剂量75 mmol/L的乙醇培养3 d、5 d、7 d后,采用免疫荧光观察MAP2、NURR1、TopoⅡβ的表达变化;Fluoro-Jade B荧光染色观察神经细胞变性坏死情况。结果:随乙醇染毒时间的延长,原代培养的神经细胞分布较正常对照组稀疏,MAP2的表达发生了变化,以7 d组变化最明显,突起变细变短,阳性表达减弱;NURR1及TopoⅡβ的表达也随乙醇染毒时间的延长,阳性细胞数目及阳性信号表达的强度均呈下降趋势,以7 d最为显著。NURR1阳性细胞计数为(9.6±3.5)与正常对照组的(45.2±3.96)有显著差异(P0.05)。TopoⅡβ(55.8±3.77)与正常对照组的(86.2±4.82)有明显差异(P0.05);Fluoro-Jade B荧光标记显示在乙醇染毒5 d组及7 d组均可见阳性细胞。结论:MAP2、NURR1、TopoⅡβ可能参与了乙醇导致的神经细胞分化异常,改变了细胞骨架稳定,最终导致了部分神经细胞的变性坏死。     

The Inhibitory Effect of Binens Bipinnata L. Extract on U14 Tumour in Mice

The objective of this paper was to study the in vitro and in vivo inhibitory effect of Bidens bipinnata L. extract on growth of cervical carcinoma U14 cells. MTT method was used to determine the inhibitory effect of Bidens bipinnata L. extract on U14 tumour cells, and the effects of Bidens bipinnata L. extract on inhibition rate of solid tumour and life prolongation rate of ascites tumour were observed through the establishment of two animal models of mouse cervical carcinoma U14 solid tumour and ascites tumour. In the in vitro MTT assay, the inhibition rate gradually increased with the increase of dose of Bidens bipinnata L. and the extension of time. Its inhibition rate was 70.44% at a concentration of 80µg/L. Solid tumour inhibition rates in the high- and low-dose groups and cisplatin group were 49.13%, 2.26% and 75.72% respectively; life prolongation rates in each ascites tumour group were 63.63%, 34.86% and 87.34% respectively. The Bidens bipinnata L. extract has a certain inhibitory effect on growth of mouse cervical carcinoma U14.     

Different activities of Schinus areira L.: anti-inflammatory or pro-inflammatory effect

The anti-inflammatory drugs possess many serious side effects at doses commonly prescribed. It is really important to discover novel regulators of inflammation from natural sources with minimal adverse effects. Schinus areira L. is a plant native from South America and is used in folk medicine as an anti-inflammatory herb. For this study, the activity of aqueous extracts on inflammation and the effect on superoxide anion production in mice macrophages were assayed. Aqueous extracts were prepared by soaking herbs in cold water (cold extract), boiling water (infusion), and simmering water (decoction). Cold extract possess an anti-inflammatory activity. Decoction and infusion showed pro-inflammatory activity. Cold extract increased the production of superoxide anion. It has been proposed to use diverse methods to obtain extracts of S. areira L. with different effects. Cold extract, decoction, and infusion could be utilized as extracts or as pharmacological preparations for topical application.     

The effect of treatment with interferon-gamma on type II collagen-induced arthritis.

We have investigated the effects of recombinant murine interferon-gamma (rIFN-gamma) on type II collagen-induced arthritis (CA) in DBA/l mice. Therapeutic as well as prophylactic treatment with subcutaneous rIFN-gamma, at 10(5) U/mouse six times a week, inhibited the development of CA without any obvious side effects. The accompanying suppression of anti-CII antibody responses may partly explain the inhibition of CA by rIFN-gamma. The possible role of the anti-inflammatory effect of systemic IFN-gamma in the inhibition of CA is discussed.     

The effect of FK506 and cyclosporin A on antigen-induced arthritis.

FK506 and cyclosporin A inhibited the development of antigen-induced arthritis in the rat and rabbit. FK506 was five times more potent than cyclosporin A in the rat and approximately 20 times more potent in the rabbit. FK506 was effective in both species if administered either from the day of intra-articular administration of antigen or when the arthritis was established. In the rabbit, arthritis returned when administration of FK506 was stopped. FK506 (10 mg/kg/day) caused renal damage which was not observed at a dose of 2.5 mg/kg/day. Both of these doses were equally effective at inhibiting the arthritis. The conclusion from these studies is that FK506 is a more effective anti-arthritic agent than cyclosporin A and that a pronounced therapeutic effect can be achieved at non-toxic doses of the drug.     

Protective effect of date palm fruit extract (Phoenix dactylifera L.) on dimethoate induced-oxidative stress in rat liver

Nowadays, people’s exposure to chemical compounds such as organophosphorus insecticides is continuously on the rise more and more. Theses compounds have induced an excessive production of free radicals which are responsible for several cell alterations in the organism. Recent investigations have proved the crucial role of nutritional antioxidants to prevent the damage caused by toxic compounds. In this study, we investigate the role of date palm fruit extract (Phoenix dactylifera L.) in protection against oxidative damage and hepatotoxicity induced by subchronic exposure to dimethoate (20 mg/kg/day). Oral administration of dimethoate caused hepatotoxicity as monitored by the increase in the levels of hepatic markers enzymes (transaminases, alkaline phosphatase, gamma-glutamyl transferase and lactate dehydrogenase), as well as in hepatic malondialdehyde thus causing drastic alteration in antioxidant defence system. Particularly, the activities of superoxide dismutase (SOD) and glutathione peroxidase (GPx) were found increased by dimethoate while catalase (CAT) activity was reduced significantly. These biochemical alterations were accompanied by histological changes marked by appearance of vacuolization, necrosis, congestion, inflammation, and enlargement of sinusoids in liver section. Pretreatment with date palm fruit extract restored the liver damage induced by dimethoate, as revealed by inhibition of hepatic lipid peroxidation, amelioration of SOD, GPx and CAT activities and improvement of histopathology changes. The present findings indicate that in vivo date palm fruit may be useful for the prevention of oxidative stress induced hepatotoxicity.     

苏木提取物及其单体成分抗排斥反应的研究进展

苏木(Caesalpinia sappan L.)为豆科云实属植物,其干燥心材为传统中药材.苏木不同提取物及其单体成分均有一定的抗排斥反应作用,但由于提取方式及其单体成分的不同,其抗排斥反应的强度及作用机制也存在着差异.苏木及其单体化合物将在自身免疫性疾病和异体移植等免疫排斥反应性疾病方面拥有广阔的前景.     

Inhibitory effect of triptolide on interleukin-18 and its receptor in rheumatoid arthritis synovial fibroblasts

OBJECTIVE: To determine the effects of triptolide (TP) on the expression of interleukin-18 (IL-18) and its receptor in phorbol 12-myristate 13-acetate (PMA)-stimulated rheumatoid arthritis synovial fibroblasts (RASF). MATERIALS AND METHODS: RASF were obtained from the synovial tissue of patients with RA. RASF were pretreated with TP (0~100 ng/ml) for 2 h before stimulation with PMA (50 ng/ml). The bioactivity of IL-18 in the supernatant was detected based on IFN-gamma secretion from IL-18-responding human myelomonocytic KG-1 cells. IL-18 level was analyzed by ELISA. In situ expression of IL-18Ralpha was determined by immunofluorescence assay. To estimate the protein and mRNA expression of IL-18 and IL-18Ralpha in RASF, western blot and quantitative RT-PCR were performed. Nuclear factor-kappaB (NF-kappaB) activity in the whole-cell extract of treated RASF was also measured using an ELISA-based method. RESULTS: TP effectively inhibited the bioactivity of IL-18 in PMA-stimulated RASF. The expression of IL-18 and IL-18R at protein and gene levels was reduced by TP. NF-kappaB activity in PMA-stimulated RASF was profoundly suppressed by TP. These effects showed a high correlation with TP concentration (0~100 ng/ml). CONCLUSION: TP effectively inhibited the expression of IL-18 and its receptor in PMA-stimulated RASF. These results suggest a mechanism of TP in RA therapy.     

山茶籽醇提取液对去卵巢阿尔茨海默病大鼠学习记忆、脑组织β淀粉样蛋白的影响

目的:观察山茶籽醇提取液对去卵巢阿尔茨海默病(AD)大鼠学习记忆、脑组织β淀粉样蛋白(Aβ)的影响,为临床应用山茶籽醇提取液防治雌激素缺乏AD提供实验依据。方法:5月龄健康雌性大鼠分成对照组、AD模型组、山茶籽治疗组和雌二醇治疗组。各实验组第4、8、12周分别取大鼠,称重后进行水迷宫实验,再麻醉大鼠取左心血5 ml,处死取大脑。免疫组织化学显色的大脑组织片用显微镜观察;右侧大脑用低温匀浆提取液进行Aβ分析。结果:AD模型组经水迷宫实验证明其学习记忆能力明显下降,脑组织海马CA1免疫组织化学显色Aβ沉积明显增加,Aβ阳性细胞计数明显增加,脑组织Aβ_(1-40)电泳带光密度值明显增加,山茶籽治疗组和雌二醇(E2)治疗组的学习记忆能力明显提高,海马CA1免疫组织化学显色Aβ沉积和Aβ阳性细胞计数明显减少,脑组织Aβ_(1-40)电泳带光密度值明显变小。结论:山茶籽醇提取液有雌激素样作用,可以减少AD模型大鼠脑组织β淀粉样蛋白沉积,提高其学习记忆能力,对去卵巢AD模型大鼠有防治作用。     

透骨草提取物诱导人宫颈癌Hela细胞凋亡及其相关机制研究

目的:探讨透骨草提取物诱导人宫颈癌Hela细胞凋亡及其相关机制。方法:荧光显微镜观察Hela细胞形态,透射电镜观察Hela细胞超微结构,流式细胞术检测Hela细胞凋亡率,Western blot检测Bcl-2蛋白和Caspase-3蛋白表达水平。结果:100μg/ml透骨草提取物处理的Hela细胞后,荧光显微镜显示Hela细胞皱缩,细胞核浓缩呈新月状边集。透射电镜可见Hella细胞表面突起和微绒毛减少,核断裂、染色质凝聚且边缘化,有"出芽"现象。流式细胞术显示透骨草提取物处理的Hela细胞凋亡率为(25.90±1.13)%,明显高于对照组(P<0.01)。Western blot结果显示透骨草提取物处理的Hela细胞Bcl-2蛋白和Caspase-3蛋白表达达明显低于与对照组(P<0.05)。结论:透骨草提取物可诱导Hela细胞凋亡,其机制可能与下调Bcl-2蛋白和Caspase-3蛋白表达有关。     

The suppressive effect of gelatin-conjugated superoxide dismutase on disease development and severity of collagen-induced arthritis in mice.

We studied the effect of superoxide dismutase (SOD) on murine collagen-induced arthritis (CIA), an animal model of human rheumatoid arthritis (RA). Among SOD derivatives studied, only gelatin-SOD conjugate which has prolonged half life in vivo was effective to suppress the development of CIA, while native SOD or gelatin carrier alone was ineffective. Interestingly, pyran polymer-conjugated SOD which also has a long half life showed no suppressive effect on the disease. No significant effect on immune response against type II collagen (CII) was found in any of the experimental groups. In addition, induction of suppressor cells was not detected in spleen or lymph node cells of the gelatin-SOD-treated group. Therefore, these results suggest that oxygen radicals may have an important role in the effector phase of the immune response to manifest this chronic autoimmune polyarthritis. Thus, the use of appropriate antioxidants for the treatment of human RA may be rationalized.     

枳椇皮提取物对泌尿结石大鼠肾功能及c-jun氨基末端激酶 信号通路的调节作用及其机制

目的：探讨枳椇皮提取物对泌尿结石大鼠肾功能及c-jun 氨基末端激酶（JNK）信号通路的调节作用及其 机制。方法：健康大鼠随机分为对照组、泌尿结石模型组（ 模型组）、枳椇皮提取物不同剂量治疗组（ 治疗组）、 排石颗粒药物对照组（ 排石组）。H-E 染色观察肾组织病理组织学形态;比色法检测血尿素氮（BUN）和肌酸酐（Cr） 水平;TUNEL 法检测肾小管上皮细胞凋亡;免疫组织化学检测肾组织p-JNK、JNK蛋白表达;黄嘌呤氧化酶法 检测肾组织超氧化物歧化酶（SOD）水平;TBA法检测肾组织丙二醛（MDA）水平。结果：模型组大鼠尿液中的 草酸钙结晶排出明显较少,低剂量组、高剂量组与排石组草酸钙结晶排出量均较模型组高,且以高剂量组效果最 佳。与对照组相比,模型组大鼠肾组织内有炎症细胞浸润,管内充满草酸钙结晶,肾小球被挤压,肾小管内有炎 性渗出物及坏死。低剂量组、高剂量组、排石组较模型组均有所减轻。与对照组相比,模型组血BUN、Cr、p-JNK、 JNK、MDA、肾小管上皮细胞凋亡升高,SOD 降低。与模型组相比,低剂量组BUN、Cr、p-JNK、JNK、MDA、 肾小管上皮细胞凋亡降低,SOD 升高。与低剂量组相比,高剂量组与排石组BUN、Cr、p-JNK、JNK、MDA、 肾小管上皮细胞凋亡降低,SOD 升高。高剂量组与排石组各指标差异无统计学意义。结论：枳椇皮提取物通过降 低JNK水平,减少了肾小管上皮细胞的凋亡,降低氧化损伤,从而改善泌尿结石大鼠肾功能,起到治疗的作用。     

Inhibitory effect of Paeonia lactiflora Pallas extract (PE) on poly (I:C)-induced immune response of epidermal keratinocytes

Epidermal keratinocytes provide protective role against external stimuli by barrier formation. In addition, kertinocytes exerts their role as the defense cells via activation of innate immunity. Disturbance of keratinocyte functions is related with skin disorders. Psoriasis is a common skin disease related with inflammatory reaction in epidermal cells. We attempted to find therapeutics for psoriasis, and found that Paeonia lactiflora Pallas extract (PE) has an inhibitory potential on poly (I:C)-induced inflammation of keratinocytes. PE significantly inhibited poly (I:C)-induced expression of crucial psoriatic cytokines, such as IL-6, IL-8, CCL20 and TNF-α, via down-regulation of NF-κB signaling pathway in human keratinocytes. In addition, PE significantly inhibited poly (I:C)-induced inflammasome activation, in terms of IL-1β and caspase-1 secretion. Finally, PE markedly inhibited poly (I:C)-increased NLRP3, an important component of inflammasome. These results indicate that PE has an inhibitory effect on poly (I:C)-induced inflammatory reaction of keratinocytes, suggesting that PE can be developed for the treatment of psoriasis.     

白扁豆多糖对免疫抑制小鼠的免疫调节作用

目的探讨白扁豆多糖(polysaccharide from Dolichos lablab L.,DLP)对环磷酰胺(cyclophosphamide,CTX)所致的免疫抑制小鼠的免疫调节作用。方法小鼠随机分为正常对照组、模型组、香菇多糖(Lentinan,LNT,15 mg/kg)和白扁豆多糖低、中、高剂量组(75、150、300 mg/kg)。通过对小鼠连续3 d腹腔注射环磷酰胺(100 mg/kg)建立小鼠免疫抑制模型,造模后连续灌胃给药7 d,检测免疫器官重量指数、血清溶血素、脾淋巴细胞增殖、脾脏NK细胞活性及小鼠血清细胞因子IL-2、IL-4和INF-γ水平,观察白扁豆多糖的免疫调节作用。结果与正常对照组相比,模型组小鼠的免疫器官重量指数、血清溶血素、脾淋巴细胞增殖、脾脏NK细胞活性及小鼠血清细胞因子IL-2、IL-4和INF-γ水平均明显降低(P<0.05,P<0.01),而中、高剂量DLP和香菇多糖可明显减轻由环磷酰胺引起的上述各项指标的下降(P<0.05,P<0.01)。结论 DLP能改善环磷酰胺所致免疫抑制小鼠的免疫功能。