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目的类风湿性关节炎(rheumatoid arthritis,RA)病程中由于炎症导致关节局部组织酸化,酸敏感离子通道(acid-sensitive ion channels,ASICs)表达与大鼠关节软骨的破坏密切相关。本研究从多途径观察苦杏仁苷对胶原诱导性关节炎(typeⅡcollagen induced arthritis,CIA)大鼠关节滑膜的免疫应答调节,探讨苦杏仁苷抗RA的作用机制。方法选取45只Wistar雌性大鼠,随机抽取8只为空白对照组。35只造模成功的CIA大鼠又随机分为模型组5只,雷公藤多甙组(即阳性对照药组)、苦杏仁苷高剂量组、苦杏仁苷低剂量组各10只。空白对照组和CIA模型组予以0.9%生理盐水,雷公藤多甙组予以雷公藤多甙1 mg/(kg·d),苦杏仁苷高剂量组予以120 mg/(kg·d),苦杏仁苷低剂量组予以60 mg/(kg·d),1次/d连续灌胃给药28 d。经腹腔麻醉,股动脉采血,常规分离血清用于IL-1β、s ICAM-1检测;分离左膝关节滑膜组织用40 g/L多聚甲醛固定作为HE染色标本;分离右膝关节滑膜组织-80℃中保存用于Western blot ASIC3蛋白检测。同时提取脑组织于-80℃中保存作为阳性对照。结果 HE染色显示CIA模型组滑膜细胞排列疏散、紊乱、增生,大量的炎细胞浸润;苦杏仁苷高剂量组、低剂量组和雷公藤多甙组滑膜轻度增生,炎性细胞浸润明显减少,外周血中IL-1β、s ICAM-1水平均有明显下降,滑膜中ASIC3蛋白表达也明显下降,与CIA模型组比较,差异有统计学意义(P<0.05);与空白组比较,差异均无统计学意义(均P>0.05)。结论苦杏仁苷能有效通过抑制CIA大鼠外周血中炎性细胞因子IL-1β、s ICAM-1的表达水平,以及下调滑膜ASIC3蛋白的表达,干扰对关节滑膜的损伤,起到抗炎和免疫抑制作用,可能是苦杏仁苷的抗炎镇痛和保护软骨作用的机制之一。
In the course of rheumatoid arthritis (RA), the local tissue acidification and the expression of acid-sensitive ion channels (ASICs) are closely related to the destruction of rat articular cartilage due to inflammation. In this study, we observed the regulatory effect of amygdalin on the synovial membrane in rats with type II collagen induced arthritis (CIA), and explored the anti-RA mechanism of amygdalin. Methods Forty-five Wistar female rats were selected and randomly selected as blank control group. Thirty-five CIA rats were randomly divided into model group (n = 5), tripterygium glycosides group (positive control group), amygdalin high dose group and amygdalin low dose group (n = 10). The blank control group and CIA model group were given 0.9% saline, tripterygium glycosides tripterygium glycosides 1 mg / (kg · d), amygdalin high dose group was 120 mg / (kg · d), bitter almond Glycosides low dose group was given 60 mg / (kg · d), 1 / d continuous oral administration of 28 d. The serum of the femoral artery was collected by intraperitoneal anesthesia and the femoral artery was collected for detection of IL-1β, s ICAM-1. The synovial tissue of left knee was isolated and fixed with 40 g / L paraformaldehyde as HE stain. Tissues were stored at -80 ° C for Western blot analysis of ASIC3 protein. At the same time, brain tissue was extracted and stored at -80 ℃ for positive control. Results HE staining showed that the synovial cells in CIA model group were evacuated, disorganized, proliferated and infiltrated with a large number of inflammatory cells. The synovial hyperplasia and infiltration of inflammatory cells in the amygdalin high dose group, low dose group and tripterygium glycosides group were obvious (P <0.05). Compared with the control group, the levels of IL-1β, s ICAM-1 in peripheral blood were significantly decreased and the expression of ASIC3 in synovial membrane was also significantly decreased , There was no significant difference (all P> 0.05). Conclusion Amygdalin can effectively inhibit the expression of inflammatory cytokines IL-1β, s ICAM-1 in peripheral blood of CIA rats and down-regulate the expression of synovial ASIC3 protein, interfere with the synovial membrane damage, Inflammation and immunosuppressive effects may be one of the mechanisms of anti-inflammatory and analgesic action of amygdalin and protection of cartilage function.