藏药独一味抗类风湿关节炎的作用及其药效物质基础研究

目的:研究藏药独一味抗类风湿关节炎(RA)的作用及其药效物质基础。方法:将56只SD大鼠随机分为正常对照组(0.5%羧甲基纤维素钠溶液),模型组(0.5%羧甲基纤维素钠溶液),甲氨蝶呤组(阳性对照组,3 mg/kg),独一味水提液低、中、高剂量组(0.5、1、2 g/kg,以生药量计)和独一味总黄酮组(200 mg/kg,以黄酮提取物计),每组8只。除正常对照组外,其余各组大鼠均于右后足垫注射弗氏完全佐剂(FCA)复制佐剂性关节炎模型。注射FCA次日,各组大鼠灌胃相应药物,每天给药1次(仅甲氨蝶呤组每3天给药1次),连续给药30天。给药第15、30天时,分别测定大鼠左后足的足肿胀度,计算其关节炎指数。第30天给药结束后,采用酶联免疫吸附法测定各组大鼠血清中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、IL-6、IL-10水平,计算其胸腺指数、脾指数,并采用苏木精-伊红染色法观察各组大鼠踝关节的病理学变化。结果:与正常对照组比较,模型组大鼠给药第15、30天时的足肿胀度、关节炎指数以及给药第30天时的脾指数和血清中TNF-α、IL-1β、IL-6水平均显著升高(P<0.01);给药第30天时的胸腺指数、IL-10水平均显著降低(P<0.01);踝关节可见明显的滑膜细胞增生、浸润关节腔等病理学改变。与模型组比较,各给药组大鼠给药第15、30天时的足肿胀度、关节炎指数以及给药第30天时的脾指数和血清中TNF-α、IL-1β、IL-6水平均显著降低(P<0.01);给药第30天时的胸腺指数和血清中IL-10水平均显著升高(P<0.01);关节炎病理学变化明显改善。与独一味水提液高剂量组比较,独一味总黄酮组大鼠给药第15天时的足肿胀度以及给药第15、30天时的关节炎指数、胸腺指数、脾指数、炎症细胞因子水平、踝关节病理学变化等差异均无统计学意义(P>0.05)。结论:藏药独一味具有良好的抗RA作用,总黄酮可能是其药效物质基础。

Study on Effects and Active Ingredients of Tibetan Medicine Lamiophlomis rotata against Rheumatoid Arthritis

OBJECTIVE:To study the effects and active ingredients of Tibetan medicine Lamiophlomis rotata against rheumatoid arthritis(RA).METHODS:Fifty-six SD rats were randomly divided into normal control group(0.5%sodium carboxymethylcellulose),model group(0.5%sodium carboxymethylcellulose),methotrexate group(positive control group,3 mg/kg),L.rotata water extract low-dose,medium-dose and high-dose groups(0.5,1,2 g/kg,by crude drug),L.rotata total flavonoid group(200 mg/kg,by flavonoid extat),with 8 rats in each group.Except for normal control group,other groups were given Freund’s complete adjuvant(FCA)on the rat’s right hind footpad to induce adjuvant-induced arthritis model.The next day after injection of FCA,rats in all groups were given relevant medicine intragastrically,once a day(once every 3 days for methotrexate group)for 30 days.At 15th and 30th day of administration,the degree of paw swelling of left hind foot was measured,and the arthritis index,spleen index were calculated.At the end of 30th day of administration,the levels of TNF-α,IL-1β,IL-6 and IL-10 in rat serum were determined by ELISA assay,the thymus index and spleen index were calculated,the pathological changes of the ankle joints were observed by HE staining.RESULTS:Compared with normal control group,degree of paw swelling,arthritis index at 15th and 30th day of administration as well as the spleen index and the levels of TNF-α,IL-1β,IL-6 in serum at 30th day of administration were significantly increased in model group(P<0.01);while the thymus index and IL-10 level at 30th day of administration were significantly decreased(P<0.01);synovial cell proliferation and infiltration of articular cavity were observed in ankle joint.Compared with model group,degree of paw swelling,arthritis index at 15th and 30th day of administration as well as the spleen index and the levels of TNF-α,IL-1β,IL-6 in serum at 30th day of administration were significantly decreased in each medicine group(P<0.01);while the thymus index and IL-10 level at 30th day of administration were significantly increased(P<0.01);the pathological changes of arthritis were significantly improved.Compared with L.rotata water extract high-dose group,there were no significant differences in degree of paw swelling at 15th day of administration as well as the arthritis index,spleen index,levels of inflammatory cytokines and pathological changes of ankle joint in L.rotata total flavonoid group(P>0.05).CONCLUSIONS:Tibetan medicine L.rotata shows well anti-RA activity,and total flavonoids may be the active ingredients of its efficacy.