淫羊藿苷拮抗脂多糖炎症模型的体内和体外研究

作者	单位
吴金峰	复旦大学附属华山医院中西医结合肺、炎症、肿瘤研究室
董竞成	复旦大学附属华山医院中西医结合肺、炎症、肿瘤研究室
徐长青	复旦大学附属华山医院中西医结合肺、炎症、肿瘤研究室
张心菊	复旦大学附属华山医院中心实验室
曾丽绚	复旦大学附属华山医院中西医结合肺、炎症、肿瘤研究室

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中文摘要:

目的从前炎症细胞因子、炎性介质、黏附分子等环节,评价淫羊藿苷的抗炎作用。方法以脂多糖(LPS)刺激小鼠巨噬细胞系RAW264.7建立体外炎症模型,以LPS刺激C57BL/6J小鼠建立体内炎症模型,采用淫羊藿苷干预,并以地塞米松为阳性对照。采用CCK-8试剂盒检测细胞活力;采用酶联免疫法检测细胞培养上清和血清中肿瘤坏死因子α(TNF-α)、白细胞介素-6(IL-6)含量;采用Griess Reagent法检测细胞培养上清一氧化氮(NO)含量;采用流式细胞术检测小鼠中性粒细胞表面黏附分子CD11b表达;采用病理切片HE染色观察肺部炎症细胞浸润。结果体外研究表明:淫羊藿苷(1、10、100μg/mL)均未显示出细胞毒性作用(P<0.01);淫羊藿苷(10、100μg/mL)可显著抑制RAW264.7TNF-α、IL-6产生(P<0.01);淫羊藿苷100μg/mL可显著抑制NO释放(P<0.01)。体内研究表明:淫羊藿苷(20mg/kg)可显著降低小鼠血清TNF-α、IL-6的水平(P<0.01),减少小鼠中性粒细胞表面黏附分子CD11b的表达(P<0.01),减轻小鼠肺部炎症细胞浸润。结论淫羊藿苷是一种安全有效的天然抗炎药物,对前炎症细胞因子(如TNF-α、IL-6)、炎症介质(如NO)和黏附分子(如CD11b)等的多环节干预是其部分疗效机理。

英文摘要:

Objective To evaluate the anti-inflammatory effects of icariin,from aspects of pro-inflammatory cytokines,inflammatory mediators and adhesion molecules.Methods Mouse inflammation model in vitro was established by stimulating macrophage cell line RAW264.7 with lipopolysaccharide(LPS);and the inflammation model in vivo was established by stimulating C57BL/6J mouse with LPS.Taking dexamethasone as the positive control,both models were treated with icariin,and the cell viability in model mice was detected with CCK-8 kit;tumor necrosis factor-α(TNF-α),interleukin-6(IL-6) in cell culture medium and serum were detected by ELISA;nitric oxide(NO) in cell culture medium by Griess Reagent method;CD11b expression on the surface of neutrophil in mice by flow cytometry,and pulmonary inflammatory cell infiltration in mice by pathological section as well.Resultsin vitro studies showed that icariin at the doses of 1 μg/mL,10 μg/mL and 100 μg/mL,all displayed no cytotoxicity(P<0.01);10 μg/mL and 100 μg/mL icariin effectively lowered the levels of TNF-α and IL-6(P<0.01) in medium;and 100 μg/mL icariin significantly reduced level of NO(P<0.01) in medium.in vivo studies showed that icariin at the dose of 20mg/kg significantly lowered serum TNF-α and IL-6 levels(P<0.01),reduced the average fluorescence intensity of adhesion molecules CD11b(P<0.01),and alleviated pulmonary inflammatory cell infiltration.ConclusionI cariin is a safe and effective natural anti-inflammatory drug,its partial mechanism is possible the multiple links intervention on pro-inflammatory cytokines(TNF-α,IL-6),inflammatory mediators(NO) and adhesion molecules(CD11b).

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