牛膝-杜仲药对对小鼠巨噬细胞RAW264.7的抗炎作用

目的研究牛膝-杜仲药对的抗炎作用。方法将小鼠巨噬细胞RAW264.7分为空白组、模型组、牛膝组(800μg/mL)、杜仲组(800μg/mL)和牛膝-杜仲药对低、中、高浓度组(400、800、1 600μg/mL),除空白组和模型组外,其余各组加入相应药物培养6 h后,空白组继续加入培养基,模型组加入10μg/mL脂多糖(以复制炎症模型),其余各组加入相应药物和10μg/mL脂多糖。检测各组细胞中炎症因子一氧化氮(NO)、白细胞介素1β(IL-1β)、IL-6、肿瘤坏死因子α(TNF-α)]的水平,并采用金氏公式评价牛膝、杜仲的联合作用;检测细胞中诱导型一氧化氮合酶(iNOS)、环氧合酶2(COX-2)、核因子κB(NF-κB)抑制蛋白α(IκBα)的表达水平和NF-κB p65、IκB激酶(IKK)、p38丝裂原激活的蛋白激酶(p38 MAPK)、胞外信号调节激酶(ERK)、c-Jun N端激酶(JNK)的磷酸化水平。结果与空白组比较,模型组细胞中炎症因子水平以及i NOS、COX-2蛋白表达水平和NF-κB p65、IKK、p38 MAPK、ERK、JNK蛋白磷酸化水平均显著升高(P<0.01),IκBα蛋白的表达水平显著降低(P<0.01);经牛膝-杜仲药对干预后,细胞中炎症因子水平以及上述蛋白的表达水平或磷酸化水平大部分显著逆转(P<0.05或P<0.01),且牛膝-杜仲药对具有协同作用。结论牛膝-杜仲药对对RAW264.7细胞具有协同抗炎作用,其作用机制可能与抑制NF-κB/MAPK信号通路相关蛋白表达有关。

Anti-inflammatory effect of couplet medicinals of Achyranthes bidentata-Eucommia ulmoides on mouse macrophage RAW264.7

OBJECTIVE To study the anti-inflammatory effect of couplet medicinals of Achyranthes bidentata-Eucommia ulmoides. METHODS Mouse macrophage RAW264.7 were divided into blank group,model group,A. bidentata group(800μg/m L),E. ulmoides group(800 μg/mL)and low-,medium-and high-concentration groups of couplet medicinals of A. bidentataE. ulmoides(400,800,1 600 μg/mL). Excep for blank group and model group,the other groups were added with corresponding drugs for 6 hours;then blank group was continued to add into the medium,while model group was added into 10 μ g/mL lipopolysaccharide(to induce the inflammatory model);other groups were added into corresponding drugs and 10 μ g/mL lipopolysaccharide. The levels of inflammatory factors nitric oxide(NO),interleukin-1β(IL-1β),IL-6,tumor necrosis factor-α(TNF-α)] were detected,and Jin’s formula was used to evaluate the effects of A. bidentata-E. ulmoides. The expression of inducible nitric oxide synthase(iNOS),cyclooxygenase-2(COX-2),nuclear factor kappa-B(NF-κB)and inhibitor α of NF-κB(IκBα)as well as the phosphorylation of NF-κB p65,IκB kinase(IKK),p38 mitogen-activated protein kinase(p38 MAPK),extracellular signal-regulated kinase(ERK)and c-Jun N-terminal kinase(JNK)were determined. RESULTS Compared with blank group,the level of inflammatory factors,protein expression of iNOS and COX-2 as well as the phosphorylation of NF-κB p65,IKK,p38 MAPK,ERK and JNK were increased significantly(P<0.01),while the protein expression of I κ B α was decreased significantly(P<0.01). After intervention of couplet medicinals of A. bidentata-E. ulmoides,the level of inflammatory factors,the expression or phosphorylation of above proteins were reversed significantly(P<0.05 or P<0.01),and couplet medicinals of A.bidentata-E. ulmoides had a synergistic effect. CONCLUSIONS The couplet medicinals of A. bidentata-E. ulmoides have synergistic anti-inflammatory effect on RAW264.7 cells. Its mechanism may be related to the inhibition of NF-κB/MAPK signaling pathway related protein expression.