清络通痹颗粒对胶原关节炎大鼠滑膜炎的影响

目的：探讨清络通痹颗粒对胶原性关节炎大鼠滑膜细胞的影响及机理。方法：建立胶原性关节炎大鼠（CIA）模型，同时设正常对照组。给药自第7天始，持续21天。取滑膜，检测滑膜细胞增殖能力和滑膜细胞分泌TNF-α的量。结论：一定剂量的清络通痹颗粒可抑制CIA大鼠滑膜细胞的过度增生，这与其调节与滑膜增生相关的细胞因子有关。     

清络通痹颗粒治疗类风湿性关节炎的分子作用机理研究

目的：探讨清络通痹颗粒治疗类风湿性关节炎的作用机理。方法：复制大鼠佐剂性关节炎模型，观察清络通痹颗粒对佐剂性关节炎大鼠IL－1、TNF－α、NO及组织病理学的影响。结果：清络通痹颗粒能明显降低佐剂性关节炎大鼠腹腔巨噬细胞产生的IL－1水平，减少佐剂性关节炎大鼠血浆TNF－α、血清NO含量，抑制滑膜细胞分泌TNF－α，并能减轻关节组织和细胞超微结构的病理损伤。结论：上述作用是清络通痹颗粒对佐剂性关节炎大鼠减轻关节病变，阻止骨质破坏发生，发展的主要机制。     

清络通痹冲剂对佐剂性关节炎大鼠的影响

通过分离、培养 3组 (治疗组、模型组、阳性组 )佐剂性关节炎大鼠膝关节滑膜细胞 ,检测其培养上清中肿瘤坏死因子 (TNF)的生物活性 ,观察清络通痹冲剂对佐剂性关节炎大鼠滑膜细胞分泌TNF的影响。此外将清络通痹冲剂作用于体外培养的人胚肺细胞 ,观察清络通痹冲剂对人胚肺细胞SOD活力的影响。结果表明 ,清络通痹冲剂能明显降低佐剂性关节炎大鼠滑膜细胞分泌的TNF(治疗组与模型组比P <0 0 5 ) ,并能明显增强人胚肺细胞SOD活力 (治疗组SOD活力与对照组相比P <0 0 5 )。     

清络通痹颗粒对胶原关节炎大鼠的保护作用及机制研究

目的:观察清络通痹颗粒(QLT)对胶原关节炎(CIA)大鼠的影响及机制。方法:建立CIA大鼠模型,观察清络通痹颗粒对CIA大鼠抗Ⅱ型胶原(CⅡ)抗体、外周血和腹腔巨噬细胞肿瘤坏死因子(TNF-α)水平、滑膜细胞凋亡和关节摄片等指标的影响。结果:清络通痹颗粒三个剂量组大鼠血清抗CⅡ抗体水平较模型组均有不同程度降低;清络通痹颗粒7.2,14.4g/kg剂量组大鼠血清和PMΦTNF-α的含量较模型组明显降低;清络通痹颗粒7.2,14.4 g/kg可明显促进CIA大鼠滑膜细胞的凋亡;CIA大鼠后肢关节摄片可见关节周围软组织肿胀明显,严重骨密度降低,关节腔间隙增宽,关节严重破坏,而清络通痹颗粒7.2,14.4g/kg给药的大鼠上述症状不同程度明显减轻。结论:一定剂量的清络通痹颗粒可调节CIA大鼠免疫功能紊乱,诱导滑膜细胞凋亡和抑制骨质破坏,其机制可能与抑制炎症性细胞因子TNF-α的过度分泌有关。     

蚁附蠲痹颗粒对佐剂性关节炎大鼠滑膜组织病理形态学的影响

目的:观察蚁附蠲痹颗粒对佐剂性关节炎大鼠滑膜组织中病理形态学的影响。方法:光镜下对佐剂性关节炎大鼠滑膜炎性细胞浸润、滑膜细胞、滑膜纤维组织增生评分,观察其组织形态学的改变。结果:佐剂性关节炎模型大鼠关节滑膜炎细胞浸润、增生明显。而蚁附蠲痹颗粒各给药组其病理改变较模型组明显减轻(P<0.05)。模型组大鼠组织形态学改变明显:关节滑膜衬里层细胞明显增厚,炎性细胞浸润,血管翳组织侵蚀骨组织等;而给药组组织形态学改变显著减轻。结论:蚁附蠲痹颗粒能有效减轻类风湿性关节炎对滑膜组织病理形态学的改变。     

蚁附蠲痹颗粒对佐剂性关节炎大鼠滑膜组织病理形态学的影响

目的：观察蚁附蠲痹颗粒对佐荆性关节炎大鼠滑膜组织中病理形态学的影响。方法：光镜下对佐剂性关节炎大鼠滑膜炎性细胞浸润、滑膜细胞、滑膜纤维组织增生评分，观察其组织形态学的改变。结果：佐剂性关节炎模型大鼠关节滑膜炎细胞浸润、增生明显。而蚁附蠲痹颗粒各给药组其病理改变较模型组明显减轻（P〈0．05）。模型组大鼠组织形态学改变明显：关节滑膜衬里层细胞明显增犀，炎性细胞浸润，血管翳组织侵蚀骨组织等；而给药组组织形态学改变显著减轻。结论：蚁附蠲痹颗粒能有效减轻类风湿性关节炎对滑膜组织病理形态学的改变。     

通痹合剂2号对胶原诱导型关节炎大鼠滑膜组织病理的影响

目的:研究通痹合剂2号对胶原诱导型关节炎(CIA)模型大鼠滑膜组织病理的影响.方法:滑膜组织常规脱水、包埋、制片、苏木精伊红染色,观察大鼠的滑膜组织病理分级积分情况,以此来判断通痹合剂2号对c队滑膜组织病理的影响.结果:通痹灵对CIA大鼠滑膜组织的滑膜血管增生、炎症细胞浸润、滑膜细胞增殖有明显抑制作用.滑膜病理总积分与模型组比较,差异非常显著性(P<0.001).结论:通痹合剂2号能有效抑制类风湿关节炎的滑膜炎症,阻断"滑膜-软骨"这一病理环节.     

清络通痹颗粒对胶原性关节炎大鼠以及血管内皮细胞增殖的影响

目的 观察清络通痹颗粒(QLT)时胶原性关节炎大鼠血液流变学,外周血IL-1表达的影响以及对血管内皮细胞增殖的影响.方法 制备胶原性关节炎大鼠模型(CIA),检测QLT对胶原性大鼠血液流变学和外周血IL-1的影响,MTT比色法检测QLT含药血清对血管内皮细胞增殖的影响.结果 QLT 7.2,14.4g/kg剂量组全血粘度及血浆黏度、外周血IL-1表达水平与模型组相比有明显降低,并且可显著抑制ECV304细胞的增殖.结论 QLT3个剂量组可不同程度降低全血黏度和血浆黏度,从而达到活血化淤的作用;抑制血管内皮细胞的增殖,从而达到抑制血管增生和血管生成,这可能是清络通痹颗粒治疗RA的重要机制.     

蚁附蠲痹颗粒对佐剂性关节炎大鼠滑膜组织中Caspase-3表达的影响

目的:观察蚁附蠲痹颗粒对佐剂性关节炎大鼠滑膜组织中Caspase-3表达的影响,探讨蚁附蠲痹颗粒治疗类风湿性关节炎的分子作用机制。方法:经计算机摄像系统采样后,采用Biomias计算机图像处理分析软件对佐剂性关节炎大鼠滑膜组织中信号强度进行分析,得出Caspase-3阳性染色面积及积分光密度值。结果:佐剂性关节炎模型组大鼠滑膜组织中Caspase-3表达(阳性总面积、积分光密度)较空白组明显的增高(P<0.05);阳性组和蚁附蠲痹颗粒高、中、低剂量组较空白组和模型组均显著增高(P<0.01)。结论:蚁附蠲痹颗粒使凋亡反应的效应者(Caspase-3)在滑膜组织中表达增强,促进细胞凋亡是其发挥治疗作用的一个重要环节。     

通痹颗粒对胶原性关节炎大鼠Wnt信号通路DKK1、β-catenin表达的影响

目的:观察通痹颗粒对胶原性关节炎(CIA)大鼠Wnt信号通路Dickkopf1(DKK1)和β-连环蛋白(β-catenin)表达的影响。方法:将40只健康SD雌性大鼠随机分为空白组、模型组、阳性药物组和通痹颗粒组。除空白组外,其余各组用牛Ⅱ型胶原诱导CIA大鼠模型。造模成功后,通痹颗粒组每日予通痹颗粒20g/kg灌胃,阳性药物组每日予雷公藤多苷片2.4mg/kg灌胃,其余组灌胃等量生理盐水,各组均每日灌胃1次,连续用药14d。比较各组大鼠治疗前后关节炎指数、关节肿胀度变化,治疗后滑膜病理学改变和血清DKK1、β-catenin表达情况。结果:治疗前和治疗第7、14天,与空白组大鼠比较,其余各组大鼠关节炎指数和关节肿胀度均显著增加(P0.01,P0.05)。治疗第7、14天,各治疗组关节炎指数和关节肿胀度均显著低于本组治疗前和模型组治疗同时期(P0.05),且治疗第14天时数值低于第7天(P0.05,P0.05)。空白组滑膜组织未增厚,滑膜细胞整齐排列,未观察到血管增生和炎症细胞浸润;模型组滑膜组织明显增厚,细胞排列紊乱,血管明显增生,大量炎症细胞浸润;各治疗组滑膜组织略微增厚,细胞排列较整齐,少量炎症细胞浸润,其中阳性药物组观察到血管轻度增生。与空白组比较,其余各组大鼠血清DKK1和β-catenin均显著升高(P0.05);与模型组比较,各治疗组大鼠血清DKK1和β-catenin水平明显降低(P0.05)。结论:通痹颗粒可以显著改善CIA大鼠关节滑膜的炎症反应,提示通过Wnt/β-catenin信号通路下调DKK1和β-catenin表达,从而防止异常血管增生及滑膜的炎症细胞浸润,可能是通痹颗粒治疗CIA的机制之一。     

Effects and mechanisms of total glucosides of paeony on adjuvant arthritis in rats

Total glucosides of paeony (TGP) is the major active constituent of Paeonia lactiflora Pall. The present study was carried out to investigate the effects of TGP on adjuvant arthritis (AA) of rat and its possible mechanisms. AA was induced by metatarsal footpad injection with complete Freund's adjuvant in male Sprague-Dawley rats. The secondary inflammatory reaction was evaluated by hind paw swelling, polyarthritis index. Activity of interleukin-1 (IL-1) was detected by Con A-induced thymocytes proliferation of C57BL/6J mice assay. The tumor necrosis factor alpha (TNFalpha), prostaglandin E(2) (PGE(2)) and cyclic adenosine monophosphate (cAMP) levels in synoviocytes were assessed by radioimmunoassay (RIA). PGE(2) receptors, EP2 and EP4, were analyzed by Western blot analysis. The level of IL-6 was measured by ELISA. Intragastric administration of TGP (50,100 mg/kg) significantly decreased secondary inflammatory reaction in AA rats. Suppressing the activity of IL-1 and TNFalpha, decreased PGE(2) and increased cAMP levels in synoviocytes of AA rats were observed after administration of TGP. In the immunoblot analysis, TGP could up-regulate the expression of EP2 and EP4. These results showed TGP significantly inhibited the progression of AA, and the inhibitory effects might be associated with its ability to mediate the level of cAMP and inhibit the production of IL-1, TNFalpha, IL-6 and PGE(2) from activated synoviocytes.     

痹痛灵颗粒对胶原诱导类风湿性关节炎大鼠原代滑膜细胞的影响

[目的]观察痹痛灵颗粒对胶原诱导类风湿性关节炎(CIA)大鼠关节原代滑膜细胞的影响。[方法]通过测量Ⅱ型胶原诱导大鼠类风湿性关节炎模型的足踝体积和关节肿胀评分,并对CIA大鼠原代滑膜细胞增殖能力的检测及滑膜细胞培养上清液IL-1β、TNF-α含量的测定,观察痹痛灵颗粒对滑膜细胞及滑膜细胞培养上清液IL-1β、TNF-α含量的影响。[结果]痹痛灵颗粒高、中剂量组大鼠滑膜细胞增殖小于模型组,与模型组相比有显著性差异;痹痛灵颗粒高、中、低剂量组与雷公藤多苷片组的细胞培养液中IL-1β、TNF-α水平明显低于模型组,有显著差异。[结论]痹痛灵颗粒通过抑制CIA大鼠滑膜细胞病理性增生,降低滑膜细胞培养液中IL-1β、TNF-α水平,对胶原诱导大鼠类风湿性关节炎产生一定的治疗作用。     

通痹灵对大鼠佐剂性关节炎滑膜细胞功能的影响

目的：研究通痹灵（ＴＢＬ）对佐剂性关节炎（ＡＡ）大鼠滑膜成纤维增殖及滑膜细胞分泌白细胞介素１（ＩＬ－１），肿瘤坏死因子α（ＴＮＦ－α）和前列腺素Ｅ２（ＰＧＥ２）的影响。     

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.     

白头翁皂苷对佐剂性关节炎大鼠FZD8表达的影响

研究中药白头翁有效成分白头翁皂苷(pulchinenoside,PULC)对佐剂性关节炎(adjuvant arthritis,AA)大鼠Frizzled(FZD)表达的影响。采用足跖注射完全弗氏佐剂制备AA大鼠,原代培养AA大鼠滑膜成纤维样滑膜细胞(fibroblast-like synoviocytes,FLS),Real-time qPCR检测PULC灌胃治疗对AA大鼠FLS FZD8表达的影响,MTT,ELISA检测FZD8敲除对FLS增殖和IL-1,IL-6,IL-8表达的影响,Real-time qPCR检测miR-375在FZD8异常表达中的作用。检测发现经PULC灌胃治疗后,AA大鼠FLS中FZD8表达明显降低,FZD8敲除后FLS增殖明显减弱,IL-1,IL-6,IL-8表达明显减少,PULC灌胃治疗后miR-375表达明显升高,上调表达的miR-375能够抑制FZD8的表达。PULC可能通过上调miR-375表达抑制FZD8的表达。     

中药熏蒸疗法对II型胶原诱导型关节炎大鼠滑膜细胞IL-1β mRNA表达的影响

目的 观察中药熏蒸疗法对II型胶原诱导型关节炎(CIA)大鼠关节滑膜细胞IL-1β mRNA表达水平的影响,以探讨中药熏蒸疗法治疗类风湿性关节炎(RA)的机理.方法 建立CIA模型,将雄性Wistar大鼠40只随机分为正常组、模型组、水熏组、低熏组、高熏组,观察各组大鼠膝关节滑膜细胞IL-1β mRNA的表达.结果 熏蒸各组大鼠滑膜细胞中IL-1β mRNA的表达量明显低于模型组,具有非常显著差异(P＜0.01);而各熏蒸组组间比较,高、低熏组明显低于水熏组,均有非常显著差异(P＜0.01);高熏组明显低于低熏组,亦具有显著差异(P＜0.01).结论 中药熏蒸疗法能明显抑制CIA大鼠滑膜细胞IL-1β mRNA表达,并推测中药熏蒸疗法抗炎机制与其抑制IL-1β的表达相关.     

Anti‐Arthritic and Antiinflammatory Effects of the Traditional Uighur Formula Kursi Caper In Vivo

Kursi Caper (KC) is a Uighur medicine based on caper which is widely used to treat arthritis and rheumatism, and preliminary studies in our laboratory showed that this traditional formula may possess potent antiinflammatory effects. This study confirms the antiinflammatory effect of KC in the adjuvant induced arthritis (AIA) model, the carrageenan and cotton‐pellet induced granuloma rat models, and further investigates in vivo the mechanism of action by measuring relevant indicators of anti‐arthritic activity. KC showed significant and dose‐dependent anti‐arthritic and antiinflammatory effects, demonstrated by reduced paw edema and arthritic scores in all animal models. Histopathological examination showed that KC reduced levels of synovial inflammatory factors in AIA rats. The overproduction of TNF‐α and IL‐1β was attenuated, and CAT, MDA and SOD levels were restored to normal in KC‐treated rats. KC also significantly reduced LPS‐induced proliferation of B lymphocytes and ConA induced proliferation of T lymphocytes in a dose‐dependent manner. Flow cytometry showed that the high dose KC‐treated group had a significantly decreased frequency of Th17 cells. This study indicates that KC can significantly attenuate arthritis and inflammation in rats by decreasing the levels of inflammatory cytokines, regulating oxidative stress, reducing lymphocyte proliferation and decreasing Th17. This supports the traditional use of KC as a potential modern therapeutic agent for the treatment of arthritis and related conditions. Copyright © 2015 John Wiley & Sons, Ltd.     

通痹灵对佐剂性关节炎大鼠滑膜细胞MCP-1 mRNA表达的影响

目的:探讨中药复方制剂通痹灵(以桂枝芍药知母汤为基础)对佐剂性关节炎大鼠滑膜细胞单核细胞趋化蛋白 1(MCP-1)mRNA表达的影响。方法:采用半定量RT-PCR法测定佐剂性关节炎大鼠滑膜细胞MCP-1 mRNA表达水平。结果:通痹灵明显抑制了佐剂性关节炎大鼠滑膜细胞MCP-1 mRNA的表达,提示通痹灵治疗类风湿性关节炎的分子机制与抑制MCP-1基因表达有关。     

野菊花总黄酮对佐剂性关节炎大鼠氧自由基代谢的影响

目的:探讨野菊花总黄酮(total flavonoids of Chrysanthemum indicum,TFC)对佛氏完全佐剂诱导的佐剂性关节炎(AA)大鼠模型的继发性肿胀作用及自由基代谢和免疫调节的影响.方法:用佛氏完全佐剂(FCA)建立大鼠的佐剂性关节炎模型,用足趾容积测量仪检测大鼠的继发性肿胀,观察TFC对其的治疗作用;采用试剂盒检测佐剂性关节炎大鼠的血清和腹腔巨噬细胞培养上清中的SOD活性、MDA含量以及NO水平;采用ConA诱导测定大鼠脾淋巴细胞增殖反应,采用小鼠脾细胞增殖法测定IL-2活性.结果:与模型组比较,TFC能改善佐剂性关节炎大鼠的继发性足趾肿胀,能降低血清和腹腔巨噬细胞中MDA和NO含量,升高SOD活性,同时促进脾淋巴细胞增殖和IL-2活性.结论:TFC可以抑制佐剂性关节炎大鼠的继发性足趾肿胀,其机制可能与其抗氧化作用和免疫调节有关.