秦艽不同配伍药对对风寒湿痹类风湿关节炎模型大鼠血清炎性因子及踝关节病理学改变的影响

Effects of Qinjiao in Different Combinations on the Content of Serum Inflammatory Cytokines and Ankle Pathology in Rats with Wind-cold-damp Rheumatoid Arthritis

目的观察秦艽不同配伍药对对风寒湿痹类风湿关节炎(RA)模型大鼠血清炎性因子及踝关节病理学改变的影响,阐释中药药性-病证-疗效之间的关联性。方法 2015年3—12月将80只SFP级SD大鼠随机分为空白组、模型组(单纯Ⅱ型胶原诱导)、病证模型组(Ⅱ型胶原诱导+风寒湿环境诱导)、阳性药组(雷公藤多苷干预)、单味秦艽组、秦艽威灵仙组(秦威组)、秦艽桑寄生组(秦桑组)和秦艽防己组(秦防组)。采用Ⅱ型胶原诱导加风寒湿环境因素刺激制备风寒湿痹RA大鼠模型,造模结束后各给药组按15 ml/kg给予对应的药液灌胃,空白组、模型组及病证模型组给予等量0.9%氯化钠溶液,连续28 d。观察各组大鼠一般状况、足跖肿胀度、关节炎指数(AI)评分变化及踝关节病理学改变情况,检测各组大鼠血清肿瘤坏死因子α(TNF-α)、白介素1β(IL-1β)水平。结果模型组、病证模型组大鼠AI评分高于空白组(P<0.05),各给药组大鼠AI评分低于模型组及病证模型组(P<0.05),秦威组大鼠AI评分低于单味秦艽组、秦桑组、秦防组(P<0.05)。病证模型组大鼠血清TNF-α水平高于空白组、模型组、阳性药组、单味秦艽组、秦威组(P<0.05),单味秦艽组、秦威组大鼠血清TNF-α水平低于秦防组(P<0.05)。模型组、病证模型组大鼠血清IL-1β水平高于空白组(P<0.05),除秦防组外的其他给药组大鼠血清IL-1β水平均低于模型组(P<0.05),各给药组大鼠血清IL-1β水平均低于病证模型组(P<0.05)。踝关节病理学显示,各给药组滑膜增生减轻,其中秦威组关节组织形态改善较好,秦防组较差。结论对于风寒湿痹RA,平温相配效果优于平平、平寒相配及单味药,实验结果符合中医临床"疗寒以热药"的治疗原则,平温相配治疗风寒湿痹RA的机制可能与其能够降低炎性因子TNF-α、IL-1β的水平,减轻滑膜增生及软骨破坏有关。

Objective To elaborate the correlation of traditional Chinese medicine（TCM） properties with syndrome differentiation and treatment effect by observing the effects of Qinjiao（Gentianae Macrophyllae Radix） in different combinations on the content of serum inflammatory cytokines and ankle pathology in rats with wind-cold-damp rheumatoid arthritis（RA）.Methods This study was conducted between March and December 2015.Eighty SPF grade SD rats were randomly divided into the blank group,collagenⅡmodel group,wind-cold-damp arthralgia model group,Tripterygium group,single Qinjiao group,Qinjiao-Weilingxian（Clematidis Radix et Rhizoma） group（hereinafter referred to as Qin-Wei group）,Qinjiao-Sangjisheng（Taxilli Herba） group（hereinafter referred to as Qin-Sang group）,Qinjiao-Fangji（Stephaniae Tetrandrae Radix） group（hereinafter referred to as Qin-Fang group）.For modeling,the collagenⅡmodel group was induced by collagenⅡemulsion,other model groups except the blank model group were induced by collageⅡ and exposed in wind-cold-damp environment.After modeling,the blank group,collagenⅡmodel group,wind-cold-damp arthralgia model group were given the same amount of 0.9% sodium chloride solution,each of the other groups was gavaged with 15 ml/kg corresponding drug solution,once a day with administration duration of 30 min,for 28 days.The changes of general condition,paw swelling and arthritis index（AI） were observed.The serum levels of TNF-α and IL-1β were determined.Pathological changes of ankle joint were detected by HE staining.Results Specifically,compared with the blank group,the collagenⅡmodel group and wind-cold-damp arthralgia model group had higher average AI scores （P〈0.05）;the average AI scores in Tripterygium group,single Qinjiao group,Qin-Wei group,Qin-Sang group,Qin-Fang group were lower than those in the collagenⅡmodel group and wind-cold-damp arthralgia model group（P〈0.05）;the average AI scores of Qin-Wei group were significantly lower than those of single Qinjiao group,Qin-Sang group and Qin-Fang group（P〈0.05）.The serum TNF-α levels in the wind-cold-damp arthralgia model group were higher than those in blank group,collageⅡmodel group,Tripterygium group,single Qinjiao group and Qin-Wei group（P〈0.05）;the serum TNF-α levels in single Qinjiao group and Qin-Wei group were lower than those in Qin-Fang group（P〈0.05）.The serum IL-1β levels in collageⅡmodel group and wind-cold-damp arthralgia model group were higher than those in the blank group（P〈0.05）;the serum IL-1β levels in Tripterygium group,single Qinjiao group,Qin-Wei group,Qin-Sang group except Qin-Fang group were lower than those in collageⅡmodel group（P〈0.05）;Tripterygium group,single Qinjiao group,Qin-Wei group,Qin-Sang group and Qin-Fang group had lower serum IL-1β levels than wind-cold-damp arthralgia model group（P〈0.05）.Pathological changes of ankle joint detected by HE staining showed that,the inflammation and pathological destruction were alleviated in Tripterygium group,single Qinjiao group,Qin-Wei group,Qin-Sang group and Qin-Fang group in varying degrees,joint tissue morphology improved better in Qin-Wei group,and pathological changes improved the poorest in Qin-Fang group.Conclusion For the wind-cold-damp RA,herb with mild property combined with that with warm property,has better effect than two kinds of herbs with mild properties,and herb with mild property combined with that with cold property,and single herb does.The experimental results are consistent with the clinical treatment principles of TCM,＆quot;treating the cold disease should use the hot medicine＆quot;.The mechanism of Qinjiao-Weilingxian compatibility having good effect on the disease may be related to its ability,which can reduce the serum TNF-α and IL-1β levels as well as synovial hyperplasia and cartilage destruction.