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目的:通过观察大鼠免疫及瘀血状态的变化从而研究黄芪桂枝五物汤对阳虚寒凝型骨关节炎大鼠的影响。方法:50只大鼠随机分为正常组,模型组,塞来昔布组(20.82 mg·kg~(-1)),黄芪桂枝五物汤7 d组和黄芪桂枝五物汤14 d组(30 g·kg~(-1))。除正常组外,各造模组大鼠分别采用冷固法加游泳、隔日给食及寒冷环境刺激的方法复制阳虚寒凝型骨关节炎模型。造模结束前最后2周,各给药组分别按6 m L·kg~(-1)给药,黄芪桂枝五物汤14 d组ig给药14 d,其余各给药组分别ig 7 d,正常组及模型组予生理盐水ig 7 d。ig结束后,分别观察各组大鼠一般状态;量取膝关节周长,计算膝关节肿胀度;苏木素-伊红(HE)染色观察大鼠下肢膝骨关节的病理变化;取大鼠血液,酶联免疫吸附测定(ELISA)检测血清液肿瘤坏死因子-α(TNF-α),白细胞介素~(-1)β(IL~(-1)β),血浆前列腺素E2(PGE2),血栓素B2(TXB2),6-酮前列腺素F1α(6-keto-PGF1α)含量,生化法检测血清超氧化物歧化酶(SOD)水平。结果:与正常组比较,模型组大鼠关机肿胀度明显升高,胶原纤维增粗、血管翳及炎性细胞浸润较为明显,大鼠血清TNF-α,IL~(-1)β,SOD及6-keto-PGF1α水平明显降低,PGE2,TXB2水平,TXB2/6-keto-PGF1α明显升高(P<0.05,P<0.01);与模型组比较,黄芪桂枝五物汤使造模后大鼠毛色变得柔亮,尾部瘀痕减轻,双下肢活动轻微受限,减少大鼠跛行;关节肿胀度降低,胶原纤维增粗、血管翳及炎性细胞浸润程度减轻,黄芪桂枝五物汤给药后使TNF-α,IL~(-1)β,SOD及6-keto-PGF1α水平升高(P<0.05),PGE2,TXB2水平,TXB2/6-keto-PGF1α降低(P<0.05)。结论:黄芪桂枝五物汤对阳虚寒凝型骨关节炎疗效确切,黄芪桂枝五物汤的作用效果与用药时间有关,其作用机制与抗炎及改善机体瘀血状态有关,并有可能通过抗炎细胞因子的调节作用影响机体瘀血状态。
OBJECTIVE: To study the effects of Huangqi Guizhi Wuwu Decoction on rats with Yang Deficiency and Cold Stasis Osteoarthritis by observing the changes of immune and blood stasis in rats. Methods: Fifty rats were randomly divided into normal group, model group, celecoxib group (20.82 mg · kg -1), Astragalus Guizhi Wuwu Decoction group and Astragalus Guizhi Wuwu Decoction for 14 days Group (30 g · kg -1). Except for the normal group, rats in each model group were respectively treated with cold-fixing method plus swimming, feeding on alternate days and stimulation with cold environment to replicate the model of Yang Deficiency and Condensing Osteoarthritis. The last two weeks before the end of modeling, each administration group was given 6 m L · kg -1, 14 d after administration of Huangqi Guizhi Wuwu Decoction for 14 days, and the other groups were given ig 7 d, Normal and model groups were given saline ig 7 d. At the end of ig, the general state of rats in each group were observed; the perimeter of knee joint was measured to calculate the swelling degree of knee joint; the pathological changes of knee joint of lower limbs were observed by hematoxylin-eosin (HE) staining; Serum levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), plasma prostaglandin E2 (PGE2), and thrombus were measured by enzyme linked immunosorbent assay (ELISA) TXB2, 6-keto-PGF1α, and the level of serum superoxide dismutase (SOD) was detected by biochemical method. Results: Compared with the normal group, the swelling degree of rats in the model group was significantly increased, the thickening of collagen fibers, the infiltration of blood vessels and inflammatory cells in the model group were more obvious. The levels of TNF - α, IL - 1, 6-keto-PGF1α, PGE2, TXB2 and TXB2 / 6-keto-PGF1α were significantly increased (P <0.05, P <0.01). Compared with the model group, Rat hair become shiny, tail stasis to reduce the activity of both lower limbs slightly limited to reduce the limp in rats; joint swelling decreased, thickening of collagen fibers, blood vessels and infiltration of inflammatory cell infiltration, Astragalus Gui Zhi Wu Wu Tang After administration, the levels of TNF-α, IL-1β, SOD and 6-keto-PGF1α were increased (P <0.05) . Conclusion: Astragalus Guizhi Wuwu Decoction has definite curative effect on yang deficiency and cold coagulation type osteoarthritis. The effect of astragalus and Guizhi Wuwu Decoction is related to the time of administration, and its mechanism is related to anti-inflammation and improving the state of blood stasis. May affect the body through the regulation of anti-inflammatory cytokines blood stasis.