痹肿消汤对实验性关节炎大鼠滑膜annexin1和aldolase A表达影响的同步研究

目的 通过中药复方痹肿消汤对实验性关节炎大鼠滑膜annexin1、aldolase A表达影响的同步研究,探讨痹肿消汤治疗类风湿关节炎(rheumatoid arthritis,RA)的作用机制.方法 采用皮下注射牛Ⅱ型胶原与完全福氏佐剂诱导SD大鼠实验性关节炎(collagen-induced arthritis,CIA)模型,并设立正常对照组.采用免疫印迹法(Western blot)同步检测正常组、模型组及痹肿消汤组大鼠在不同时间点滑膜annexin1、aldolase A的表达,运用ImageTool 3.0灰度扫描软件对结果进行半定量分析.结果 造模25 d后,模型组滑膜annexin 1表达低于正常组(P＜0.05),aldolase A表达高于正常组(P＜0.05).随着免疫时间的延长,模型组滑膜annexin 1表达呈水平递减趋势(P＜0.05),aldolase A表达则逐渐递增(P＜0.05).痹肿消汤治疗后,annexin 1表达在各时间点明显均高于模型组(P＜0.05),表达呈递增趋势(P＜0.05),aldolase A表达则明显低于同时间模型组(P＜0.05),在25、35和45 d水平,表达呈递减趋势(P＜0.05).BZXD能上调CIA大鼠滑膜annexin 1表达、下调aldolase A表达.结论 在CIA病理过程中,随着关节炎症状加重,annexin 1表达下调,aldolase A表达上调.提示annexin 1、aldolase A与RA滑膜病变(关节炎症、骨质侵蚀)密切相关;BZXD上调CIA大鼠滑膜annexin 1表达、下调aldolase A表达,这可能是其阻抑RA关节炎症、骨质侵蚀的重要机制之一.     

Expression and localization analyses of the cholinergic anti-inflammatory pathway and α7nAchR in different tissues of rats with rheumatoid arthritis

Rheumatoid arthritis (RA) is a complicated chronic multisystem autoimmune disease, wherein the inflammatory cascade leads to vasospasm and osteoclastogenesis, which ultimately results in bone and cartilage destruction. In this study, we investigated the expression and localization of the alpha-7 nicotinic receptor (α7nAchR) gene CHRNA7 in the heart, liver, spleen, lung, kidney, and joints of the collagen-induced arthritis (CIA) rat model. The CHRNA7 mRNA and protein expression levels in these tissues of rats from CIA and normal groups were analyzed via real-time quantitative polymerase chain reaction (RT-qPCR) and western blotting, respectively. The cellular localization of CHRNA7 protein was determined via immunohistochemistry (IHC) assays. CHRNA7 was expressed at varying levels in different tissues of rats from the groups, among which joints showed significantly higher CHRNA7 expression levels than other tissues (P < 0.05). CIA rats had significantly higher CHRNA7 expression levels in the spleen and joints than the control group rats (P < 0.05). Positive expression signals for CHRNA7 were detected in various tissues of CIA and control group rats, among which strong positive signals were detected in joint fibroblast-like synoviocytes (FLSs), endothelial cells, stromal cells, and macrophages. Our results further confirmed the involvement of the CAP in the onset and development of inflammatory responses in RA, suggesting that CHRNA7 may be a new therapeutic target for RA. This study is of great clinical and theoretical significance for understanding the differential expression of CHRNA7 in various tissues and cholinergic anti-inflammatory pathway (CAP)-targeted treatment of RA.     

卡介苗在关节炎模型小鼠诱导中的作用

目的：探讨在牛Ⅱ型胶原诱导性关节炎模型（ Collagen-induced arthritis ,CIA）小鼠的诱导过程中,弗氏完全佐剂（ FCA）中的卡介苗（ BCG）浓度和CIA小鼠关节炎严重程度的关系。方法：将牛Ⅱ型胶原蛋白溶液与含不同BCG的FCA等体积混合乳化,在小鼠尾根部注射进行免疫。免疫后观察小鼠的相关指标包括体重、发病率、血清TNF-α、关节炎指数及小鼠关节和脾脏的病理变化。结果：与1 mg/ml卡介苗组相比,4 mg/ml卡介苗组诱导的CIA小鼠发病早,关节炎指数、关节和脾脏病理评分升高及小鼠体重下降更明显,血清TGF-α明显升高,差异有统计学意义。结论：在进行CIA小鼠诱导时,BCG浓度和CIA小鼠关节炎严重程度呈正相关,实验中可以根据对动物的要求,选择含不同BCG的佐剂来诱导CIA小鼠。     

The long-term immunosuppressive effects of disulfide-linked HLA-G dimer in mice with collagen-induced arthritis

HLA-G, a natural immunosuppressant present in the human placenta during pregnancy, prevents fetal destruction by the maternal immune system. The immunosuppressive effect of HLA-G is mediated by the immune cell inhibitory receptors, LILRB1 and LILRB2. HLA-G forms disulfide-linked dimers by natural oxidation, and the dimer associates with LILRB1/B2 much more strongly than the monomer. Furthermore, the dimer formation remarkably enhanced the LILRB-mediated signaling. In this report, we studied the in vivo immunosuppressive effect of the HLA-G dimer, using the collagen-induced arthritis model mouse. Mice were treated with the HLA-G monomer or dimer intracutaneously at the left foot joint, once or for 5 days, and the clinical severity was evaluated daily in a double-blind study. The HLA-G monomer and dimer both produced excellent anti-inflammatory effects with a single, local administration. Notably, as compared to the monomer, the dimer exhibited significant immunosuppressive effects at lower concentrations, which persisted for about two months. In accordance with this result, a binding study revealed that the HLA-G dimer binds PIR-B, the mouse homolog of the LILRBs, with higher affinity and avidity than the monomer. The HLA-G dimer is expected to be quite useful as an anti-rheumatoid arthritis agent, in small amounts with minimal side effects.     

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。     

宣发膜原法对胶原诱导性关节炎大鼠免疫功能的影响

目的：探讨宣发膜原法对Ⅱ型胶原诱导性关节炎（CIA）的治疗机理，为研究类风湿关节炎（RA）的发病和损伤机理提供实验依据。方法：以CIA作为RA的动物模型，给以宣发膜原方治疗，检测CIA大鼠免疫指标和病理变化。结果：治疗组大鼠病理损伤减轻，TNF-α降低，IFN-γ升高，抗CⅡ抗体降低。结论：宣发膜原法能够减轻CIA病理损伤，调节CIA大鼠免疫功能，可以用于RA治疗。     

β-内啡肽对胶原诱导关节炎大鼠的免疫调节作用

目的研究β-内啡肽(β-END)对胶原诱导性关节炎(CIA)大鼠的免疫调节作用,为探索β-END治疗类风湿关节炎(RA)提供实验依据。方法采用尾根部皮内多点注射天然Ⅱ型胶原(CⅡ)的方法免疫雌性Wistar大鼠(60只),建立CIA模型。随机取CIA成模大鼠(5只/组)于初次免疫后第14~35天,给予不同浓度的β-END腹腔内注射,定期进行临床、实验室、影像学及病理指标评估。结果不同剂量β-END(0.1、1、5 nmol隔日1次共2周)治疗后CIA大鼠临床、实验室、影像学及病理指标明显缓解;正常鼠β-END给药5 nmol×2周后重要脏器功能、组织学未见明显异常。结论生理浓度的β-END体内可缓解CIA鼠关节局部及全身免疫炎性反应,这使β-END成为有潜力的治疗RA的制剂。     

外源性LTB4对CIA小鼠Treg/Th17脾细胞分化的作用

目的探讨外源性白三烯B4(LTB4)对胶原诱导型关节炎(collagen-induced arthritis,CIA)小鼠脾细胞调节性T细胞(Treg)和Th17细胞分化的调节,进一步阐明LTB4在类风湿关节炎(RA)发病中的作用机制。方法建立CIA小鼠模型,取造模d28的脾细胞,体外实验分析外源性LTB4对Treg和Th17细胞分化的影响。应用流式细胞术检测CD4+CD25+Foxp3+细胞的数量,荧光定量PCR技术检测调控Treg和Th17细胞分化的特异性转录因子Foxp3和RORγt的mRNA的表达,酶联免疫吸附(ELISA)方法检测培养细胞上清IL-17的含量。结果成功建立CIA小鼠模型;造模d28分离小鼠脾细胞,体外培养加入鸡Ⅱ型胶原(CⅡ)共同孵育,随着LTB4浓度增加(0.01、0.1、1μmol·L-1),CD4+CD25+Foxp3+细胞数量相应减少,Foxp3 mRNA的表达相应降低;相反,IL-17的含量相应增加,RORγt mRNA的表达相应升高。结论LTB4抑制CIA模型脾细胞Treg细胞的分化,促进Th17细胞的分化,提示LTB4在CIA发病过程中具有一定的促炎活性。     

Artocarpus: A review of its traditional uses,phytochemistry and pharmacology

The genus Artocarpus (Moraceae) comprises about 50 species of evergreen and deciduous trees. Economically, the genus is of appreciable importance as a source of edible fruit, yield fairly good timber and is widely used in folk medicines. The aim of the present review is to present comprehensive information of the chemical constituents, biological and pharmacological research on Artocarpus which will be presented and critically evaluated. The close connection between traditional and modern sources for ethnopharmacological uses of Artocarpus species, especially for treatment against inflammation, malarial fever, diarrhoea, diabetes and tapeworm infection. Artocarpus species are rich in phenolic compounds including flavonoids, stilbenoids, arylbenzofurons and Jacalin, a lectin. The extracts and metabolites of Artocarpus particularly those from leaves, bark, stem and fruit possess several useful bioactive compounds and recently additional data are available on exploitation of these compounds in the various biological activities including antibacterial, antitubercular, antiviral, antifungal, antiplatelet, antiarthritic, tyrosinase inhibitory and cytotoxicity. Several pharmacological studies of the natural products from Artocarpus have conclusively established their mode of action in treatment of various diseases and other health benefits. Jacalin, a lectin present in seeds of this plant has a wide range of activities. Strong interdisciplinary programmes that incorporate conventional and new technologies will be critical for the future development of Artocarpus as a promising source of medicinal products. In the present review, attempts on the important findings have been made on identification; synthesis and bioactivity of metabolites present in Artocarpus which have been highlighted along with the current trends in research on Artocarpus.     

血管活性肠肽抑制实验性类风湿性关节炎的研究

类风湿性关节炎是一种自身抗原未明的慢性自身免疫病,以多关节的慢性炎症及渐进性骨和软骨的破坏为主要特征。胶原诱导的关节炎因其临床表现、病理组织学改变以及免疫学表现等方面与类风湿性关节炎的相似性而成为理想的动物模型。在成功建立了胶原诱导的大鼠关节炎模型(CIA)的基础上采用了血管活性肠肽(VIP)注射,研究其干预关节炎发生的效果。结果显示:使用血管活性肠肽组的大鼠CIA发病率和严重程度明显降低,关节肿胀以及骨和软骨的破坏减轻,大鼠血清中抗胶原抗体水平显著降低,大鼠T淋巴细胞对胶原的增殖反应也显著降低。提示VIP可能通过减轻对胶原的免疫应答而抑制关节炎症状,具有可能的临床应用价值。