摘要
目的:通过足跖部注射牛Ⅱ型胶原和不完全氟氏佐剂乳化剂,建立类风湿性关节炎(rheumatoid arthritis,RA)的胶原诱导型关节炎(collagen induced arthritis,CIA)模型,用相应指标观察虎杖大黄素干预炎症和新生血管形成情况。方法:Wistar大鼠40只随机分为2组,正常组,模型组,模型组采用等量的牛Ⅱ型胶原和不完全氟氏佐剂乳化后在雄性Wistar大鼠足跖部皮内注射0.4 m L(含胶原400μg),14 d后同样方法于足跖部皮内注射0.1 m L加强,致炎20 d成模后,再将模型组分为模型、虎杖大黄素与醋酸地塞米松组,每组10只,此时虎杖大黄素组0.4 g·L-1虎杖大黄素0.8 mg·kg-1ig,醋酸地塞米松组用0.75 g·L-1醋酸地塞米松7.5 mg·kg-1ig,两药各自连用20 d,并用足跖厚度,足爪X射线,足爪大体图片,膝关节滑膜HE染色,ELISA检测大鼠血清中肿瘤坏死因子-α(TNF-α)含量,免疫组化检测膝关节滑膜TNF-α及CD34表达,观察成模大鼠的2个靶点变化。结果:与正常组比较,模型组类风湿性关节炎大鼠足跖厚度明显增加(P<0.01),大鼠血清TNF-α含量明显升高(P<0.01);给药后,与模型组比较,虎杖大黄素与醋酸地塞米松组明显降低大鼠足跖厚度,血清TNF-α含量(P<0.05,P<0.01);X射线结果显示,模型组大鼠足跖部出现不连续的低密度影,即为溶骨现象,且有明显的软组织肿胀阴影,而正常组、虎杖大黄素组、醋酸地塞米松组未见有不连续的低密度影,即未见溶骨现象,亦未见软组织肿胀阴影;HE染色显示,模型组大鼠膝关节滑膜可见有大量的炎症细胞浸润和大量的血管翳,而正常组未见炎症细胞浸润和血管翳,虎杖大黄素组和醋酸地塞米松组只见少量炎症细胞浸润和少量的血管翳;免疫组化结果显示,模型组膝关节滑膜中的CD34及TNF-α的表达明显增高,虎杖大黄素组和醋酸地塞米松组膝关节滑膜中的CD34及TNF-α的表达明显降低。结论:虎杖大黄素可通过影响炎症和新生血管形成两个靶点干预RA。
Objective： To establish a collagen-induced arthritis（ CIA） model of rheumatoid arthritis（ RA） through plantar injection of bovine collagen Ⅱ and incomplete Freunds Adjuvant,in order to observe the effect of emodin from Polygoni Cuspidati Rhizoma et Radix（ PCRR） on inflammation and neovascularization with corresponding indicators. Method： Wistar rats were randomly divided into 2 groups： the normal group and the model group. The model group was given the same amounts of bovine collagen Ⅱ and incomplete freunds adjuvant and then intradermally injected with 0. 4 m L（ containing 400 μg of collagen） of solution through plantar region.After 14 days,the same method（ 0. 1 m L） was adopted for the plantar intradermal injection of additionally 0. 1 m L of solution to induce inflammation and establish the model 20 days later. After the modeling,the model group was divided into the model subgroup,the emodin from PCRR subgroup and the dexamethasone acetate subgroup,with10 in each group. Subsequently,the modin from PCRR subgroup was orally given 0. 4 g·L- 1emodin from PCRR（ 0. 8 mg·kg- 1,ig）,while the dexamethasone acetate subgroup was orally given 0. 75 g·L- 1dexamethasone acetate（ 7. 5 mg·kg- 1,ig）. The two drugs were continuously used separately for 20 d,and the plantar thickness,claw X-ray,claws gross pictures,synovial HE staining of knee joint and ELISA were used for detecting rat serum tumor necrosis factor alpha（ TNF-α） in serum. The immunohistochemistry was used to detect TNF-α and CD34 expressions of knee joint synovium,and the changes in the two targets were observed in modeled rats. Result：Compared with the normal group,the model group show significant increases in plantar thickness of RA rats（ P〈0. 01） and TNF-α content in rat serum（ P〈0. 01）,after the administration,compared with the model subgroup,the emodin from PCRR subgroup and the dexamethasone acetate subgroup showed notable decreases in plantar thickness and TNF-α content in serum（ P〈0. 05,P〈0. 01）. According to X-ray result,the model subgroup showed discontinuous low-density shadows in plantar region,which were actually notable soft tissue swelling and suggested osteolysis phenomenon,and the normal group,the emodin from PCRR subgroup and the dexamethasone acetate subgroup showed no discontinuous low-density shadows,which suggested no notable soft tissue swelling and osteolysis phenomenon. According to the HE staining,the model subgroup showed plenty of inflammatory cell infiltration and pannus at knee joint synovium,the normal group showed no inflammatory cell infiltration and pannus,and the emodin from PCRR subgroup and the dexamethasone acetate subgroup showed only a little inflammatory cell infiltration and pannus. According to the immunohistochemical result,the model subgroup showed remarkable increases in TNF-α and CD34 expressions at knee joint synovium,whereas the emodin from PCRR subgroup and the dexamethasone acetate subgroup showed significant decreases in TNF-α and CD34 expressions.Conclusion： Emodin from PCRR can intervene RA by regulating two targets TNF-α and CD34 for the inflammation and neovascularization.
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2015年第17期111-115,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81460671)
贵州省科学技术厅贵阳中医学院联合基金项目(黔科合中药字[2011]LK27043号)
关键词
类风湿性关节炎
虎杖
大黄素
炎症
新生血管形成
rheumatoid arthritis
Polygoni Cuspidati Rhizoma et Radix
emodin
inflammation
neovascularization
