【作者】 沈昆金；

【导师】 汪悦；

【作者基本信息】 南京中医药大学 ， 中医内科学， 2009， 博士

【摘要】 背景目的:类风湿性关节炎(rheumatoid arthritis)是一种以关节滑膜炎为特征的慢性全身性自身免疫性自身免疫性疾病。滑膜炎持久反复发作,可导致关节内软骨和骨的破坏,关节功能障碍。本病发病率较高,且发病以青壮年为多。本病预后欠佳,致残废率高,严重危害广大人民的身体健康。目前无论采取何种治疗方案,虽可改善症状、减轻炎症,但最终均未能有效地防止骨侵蚀,阻断病程进展,且长期应用副作用大。到目前为止,西医药治疗本病仍无较为理想的方法。本病属于中医学“痹证”、“历节”、“鹤膝风”、“尪痹”范畴。中医药治疗本病一般以辨证论治为主,通过多层次、多环节、多途径、多靶点作用于人体,抗炎镇痛,调节免疫功能,阻止病变发展,具有较大优势。本课题以古方为基础,结合全国名老中医汪履秋老先生和导师汪悦教授的经验而成热痹清,主要由石膏、知母、苍术、桂枝、银花、秦艽、虎杖、苡仁、青风藤等药组成。前期临床用于类风湿性关节炎湿热证取得了较好的疗效。本课题将对该方进一步做实验研究,观察其主要的药理药效,并初步探讨其作用机理。方法:1.抗炎作用试验:腹腔注射醋酸所致小鼠腹腔毛细血管通透性增高的影响、对巴豆油混合致炎剂所致小鼠耳廓肿胀的影响以及对大鼠塑料环肉芽肿影响试验观察其抗炎作用。2.镇痛试验:热板法、醋酸扭体法观察其镇痛作用。3.对Ⅱ型胶原关节炎大鼠的影响:观察痹痛清颗粒对大鼠Ⅱ型胶原关节炎评分、大鼠造模足关节体积、大鼠关节肿胀度及对组织病理学评分的影响。4.对Ⅱ型胶原关节炎大鼠血清细胞因子的影响:CIA小鼠滑膜细胞的分离、培养与鉴定,CIA滑膜细胞培养上清液IL-1β、TNF-α含量的测定。结果:1.抗炎作用试验:(1)腹腔注射醋酸所致小鼠腹腔毛细血管通透性增高的影响:痹痛清颗粒各剂量组腹腔液OD值均小于模型组,与模型组相比,有显著性差异。(2)对巴豆油混合致炎剂所致小鼠耳廓肿胀的影响:痹痛清颗粒各个剂量组致炎耳质量差值小于空白对照组,与空白对照组相比,有显著性差异。(3)对大鼠塑料环肉芽肿的影响:痹痛清颗粒高、中两个剂量组肉芽肿湿重和干重均小于模型,与模型组之相比,均有显著性差异。2.镇痛试验:(1)热板法:各给药剂量组在给药后各时间点痛阈提高百分率均高于空白对照组,其中0.5h时痹痛清颗粒1组、2组与对照组相比,有显著性差异。各剂量组在给药1h时,与空白对照组相比,均有显著性差异。在给药2h时痹痛清颗粒1组、2组与空白对照组相比,有非常显著性差异。(2)醋酸扭体法:痹痛清颗粒1、2二个剂量组能明显减少醋酸所致小鼠扭体反应次数,与空白对照组比较,差别有显著性差异。3.对Ⅱ型胶原关节炎大鼠的影响:痹痛清低、高剂量组和雷公藤多苷片组动物关节炎评分均低于模型组。痹痛清高、中、低剂量组,雷公藤多苷片左足和右足体积均分别小于模型组左足和右足体积;痹痛清高、中剂量组关节肿胀度均小于模型组;雷公藤片组、痹痛清颗粒高、中、低剂量组关节病变程度评分均低于模型组,其中雷公藤片组和痹痛清颗粒高剂量组与模型组相比均有显著性差异。4.对Ⅱ型胶原关节炎大鼠血清细胞因子的影响:痹痛清三个剂量组小鼠滑膜细胞培养上清液IL-1β、TNF-α含量均低于模型组,与模型组相比,具有显著性差异。结论:1.抗炎作用试验:痹痛清颗粒能抑制HAc所引起小鼠腹腔毛细血管通透性的增高;抑制巴豆油所致小鼠耳廓肿胀;抗肉芽组织形成。说明痹痛清颗粒对急性炎症反应和慢性炎症增生作用具有一定的抑制作用。2.镇痛试验:痹痛清颗粒给药后痛阈提高百分率显著增高,醋酸所致小鼠反应次数明显减少。说明痹痛清颗粒具有一定的镇痛作用。3.对Ⅱ型胶原关节炎大鼠的影响:痹痛清颗粒给药后Ⅱ型胶原关节炎大鼠的病变均有不同程度的减轻。4.对Ⅱ型胶原关节炎大鼠关节滑膜细胞培养分泌IL-1β、TNF-α的影响:结果表明,痹痛清颗粒有抑制Ⅱ型胶原关节炎大鼠关节滑膜的病理性分泌IL-1β、TNF-α。更多还原

【Abstract】 Objective:Rheumatoid Arthritis(RA) is a kind of chronic multisystematically inflammatory autoimmune disease,marked by Arthromeningitis.Because of its high incidence of disease and rate of disability,it has been one of the international tackled problems.Due to its uncertain pathogenic pathogenic cause and pathogenesis,there is no specific and pertinent drug,moreover clinical anti-rheumatic drugs in western medicine have always a lot of side effects.However,Chinese medicine is mainly characterized by bianzheng lunzhi.It has great advantage to preventing the development of disease through multi-layer,multi-loop,multi-method and multi-target way.The research lies on the clinical experience of treating active stage of tutor and mine.To explore the law of syndrome differentiation and treatment on active stage of RA and the mechanism of action of the therapeutic method through animal experiments and clinical trial.Methods:There are four parts as follows.①Establish adjuvant induced Arthritis rats model,observe Bitong Qing granule for its Arthritis evaluation,arthroncus degree and so on the effects of histopathologic change on ankle joint.②EstablishⅡcollagen induced Arthritis of rats model,observe Bitong Qing granule for its Arthritis evaluation,splenocytes proliferation response,sensitized adhesive capacity of T spleniclym phocvtes and so on the effects of synoviocytes cultured supernatant on IL-1β, TNF-αcontained quantity.③The pain threshold and frequency of body writhing were studied across a range of stimulus intensities in thermal(hot plate) and chemical irritant(acetic acid glacial) tests.④The anti-inflammatory effect was tested by ear swollen method and capillary penetration method in mice.Results:①Bitong Qing granule can obviously reduce adjuvant induced arthritis rats local paw swelling degree,histopathologic change on ankle joint,lower arthritis evaluation.②Bitong Qing granule can obviously reduceⅡcollagen induced arthritis of rats and delay the incidence of Arthritis,obviously inhibit the claws swelling and swelling evaluation scores:inhibit the T lymphocyte proliferation:inhibit an sensitized an sensitized adhesive capacity(adhesion) of T splenocytes:lower synoviocytes cultured supernatant IL-1β,TNF-αlevels.③Bitong Qing granule can prolong the pain threshold of mice and inhibit frequency of body writhing.④Bitong Qing granule can inhibite xylene-induced ear swelling and reduce acetic acid-induced permeability of the capillaries.Conclusion:Bitong Qing granule can that is basic prescription of warming meridian to expel cold associated with dispersing heat has good clinical effect on active stage of RA.The mechanism of curing RA may be in connection with the functions of anti-inflammatory,analgesia and immune regulation.更多还原