豹皮樟总黄酮对大鼠佐剂性关节炎的作用及部分机制研究

Effect of total flavonoids from Litsea coreana Leve. on adjuvant-induced arthritis and its mechanism

目的研究豹皮樟总黄酮(total flavonoids of Litseacoreana Leve,TFLC)对佐剂性关节炎的治疗作用及其机制。方法采用福氏完全佐剂诱导大鼠佐剂性关节炎(adjuvant-induced arthritis,AA)模型,观察TFLC的抗炎作用;放免法测定AA大鼠腹腔巨噬细胞(peritoneal macrophages,PMΦ)中肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白细胞介素6(interleukin-6,IL-6)的含量;MTT法检测AA大鼠脾淋巴细胞增殖反应;HE染色法观察AA大鼠膝关节病理组织学变化;免疫组化法观察AA大鼠膝关节滑膜细胞中基质金属蛋白酶9(matrix metalloproteinases,MMP-9)蛋白表达情况。结果TFLC(50、100、200mg.kg-1)对AA大鼠的原发性、继发性炎症具有明显的抑制作用,降低多发性关节炎评分;TFLC(50、100、200mg.kg-1)可提高AA大鼠低下的脾淋巴细胞增殖反应;TFLC(50、100、200mg.kg-1)有效降低AA大鼠PMΦ产生的TNF-α、IL-6水平;TFLC(50、100、200mg.kg-1)改善AA大鼠膝关节病理损伤;TFLC(50、100、200mg.kg-1)能明显降低AA大鼠膝关节滑膜细胞中MMP-9的蛋白表达。结论TFLC(50、100、200mg.kg-1)对AA大鼠具有明显的治疗作用,抗炎机制可能与调节免疫功能、减少细胞因子的生成以及抑制基质金属蛋白酶的蛋白表达有关。

Aim To investigate the effect of total flavonoids of Litsea coreana Leve. （TFLC） on adjuvant-induced arthritis （AA） and its mechanism. Method AA was induced with Feund＇s complete adjuvant; The degree of paw swelling and polyarthritis index were calculated. TNF-α and interleukin （IL-6） in peritoneal macrophages （PMΦ） of secondary arthritis rats were measured by radioimmunoassay （RIA）. ConA or LPS- induced splenocyte proliferation on AA rats was assayed with MTT reagent. Paraffin sections of knee joint stained with hematoxylin and eosin （HE） were prepared for histological analysis to assess joint damage. Protein expression of matrix metalloproteinases （ MMP- 9） on knee joint synoviocytes of AA rats was surveyed by immunohistochemistry. Results TFLC （50, 100, 200 mg·kg^-1） significantly inhibited the primary and secondary paw swelling, and decreased polyarthritis index. The splenic lymphocyte proliferation reaction of AA rats was increased by the treatment with TFLC （50, 100,200 mg·kg^-1）. TFLC （50, 100,200 mg·kg^-1 ） markedly reduced TNF-α and IL-6 level in PMΦ of AA rats. The pathological damage of knee joint was reversed by TFLC （50, 100, 200 mg·kg^-1 ）. TFLC （50, 100,200 mg·kg^-1） significantly inhibited the expression of protein MMP-9 on knee joint synoviocytes of AA rats. Conclusion TFLC （ 50, 100, 200 mg·kg^-1 ） has marked therapeutic effect on AA rats, which is related to regulating the immune function, reducing the production of cytokines and inhibiting the expression of protein MMP-9.