通痹四味方对佐剂性关节炎及体内免疫调节的作用

Effect of a Traditional Chinese Medicine Effective Parts Combination,Tongbi Siwei Formula on Treating Adjuvant Arthritis in Rats and in vivo Immunomodulation

目的观察中药有效部位组方通痹四味方(TBSWF)对大鼠佐剂性关节炎(AA)的治疗作用及小鼠体内免疫调节作用。方法采用注射弗氏完全佐剂(FCA,5 mg·mL^(-1))0.05 mL进行AA大鼠造模,造模成功的大鼠随机分为雷公藤多苷组(LGT,40 mg·kg^(-1)),腰痛宁组(YTN,150 mg·kg^(-1)),腰痹通组(YBT,380 mg·kg^(-1))及TBSWF高、中、低剂量组(140,70,35 mg·kg^(-1))。考察AA大鼠关节炎指数(AI)、足跖肿胀度及胸腺、脾脏系数等;采用酶联免疫吸附法(ELISA)测定AA大鼠血清中的前列腺素E_2(PGE_2)、肿瘤坏死因子(TNF-α)及白细胞介素1β(IL-1β)的水平。采用小鼠皮肤迟发性过敏反应模型和炭粒廓清模型分别考察TBSWF对小鼠特异性免疫和非特异性免疫的调节效果。结果与模型组比较,TBSWF高剂量组AA大鼠继发肿胀度、AI值显著降低(P<0.05,P<0.01),且抑制继发性肿胀的效果优于YTN组和YBT组(P<0.05);TBSWF高、中剂量组的大鼠体质量增加值明显增高,而脾脏系数、胸腺系数明显降低,差异有统计学意义(P<0.01,P<0.05);TBSWF高、中、低剂量组大鼠血清中PGE_2、IL-1β的含量明显降低(P<0.01),TBSWF高、中剂量组大鼠血清中TNF-α水平明显降低(P<0.01),且显著低于YTN组和YBT组(P<0.01)。小鼠皮肤迟发性过敏反应中,与空白组比较,TBSWF高、中、低剂量组的耳廓肿胀度均明显降低(P<0.01,P<0.05),且TBSWF高剂量组的脾脏系数亦明显降低(P<0.05);小鼠炭粒廓清实验中,TBSWF高、中、低剂量组小鼠的炭粒廓清指数K和校正廓清指数α均有一定程度的下降,但差异无统计学意义(P>0.05)。结论 TBSWF能抑制AA模型大鼠的继发性足跖肿胀和关节炎病变,改善大鼠机体功能,降低AA大鼠血清中炎症细胞因子水平,并对正常小鼠特异性免疫有一定的抑制作用,而对非特异性免疫抑制作用不明显。

Objective To investigate the in vivo effects of Tongbi Siwei Formula（TBSWF）, a candidate Chinese medicine formula for treating rheumatism disease, on curing the rat adjuvant arthritis （AA） and the immunomodulatory effects. Methods Complete Freund＇s Adjuvant （FCA, 0.05 mL, 5 mg·mL^-1） was used to establish rat AA model by injection, then, the modeled animals were divided into Leigongteng （LGT） group, Yaotongning （YTN） group, Yaobitong （YBT） group, TBSWF high, medium, and low dose groups （140, 70, 35 mg·kg^-1）. The arthritis index （AI） scores, the swelling of voix pedis and the index of spleen and thymus of AA rats were measured; the level of PGE2, TNF-α, IL-1β in rat plasma was determined by ELISA. Moreover, the mice skin delayed style hypersensitivity （DTH） reaction model and the mice carbon particles clearance model were applied to investigate the specific and nonspecific immunomodulatory action of TBSWF, respectively. Results Compared to model group, secondary swelling and arthritis degrees of AA rats are significantly inhibited in high dose group of TBSWF （P 〈 0.05, P 〈 0.01） , for which the effects on inhibiting secondary swelling are superior to YTN and YBT groups; the weight of AA rats was significantly increased （P 〈 0.05, P 〈 0.01 ） and the index of spleen and thymusin was remarkably decreased （P 〈 0.05, P 〈 0.01） in the high and medium dose groups of TBSWF; the levels of PGE2, IL-1β and TNF-α in TBSWF groups were significantly reduced and significantly lower than those in YTN and YBT groups （P 〈 0.01 ）. In induced mouse skin DTH reaction, auricular swelling of TBSWF group significantly decreased （P 〈 0.05, P 〈 0.01） with prominently dropping of spleen index （P 〈 0.05） compared with the blank group. In rat carbon particles clearance model, index K and index cL in TBSWF group prominently decreased, whereas TBSWF groups had no significant differences with blank group （P 〉 0.05）. Conclusion TBSWF shows a good performance in treating secondary swelling and lesion of AA rats as well as improving AA rats＇ body functions; in addition, TBSWF might show some inhibitory effect on the specific immune of normal mice, and show no obvious effect on the non-specific immunosuppression, demonstrating its good selective immunomodulatory effects.