淫羊藿素对LPS诱导原代皮质星形胶质细胞炎性反应影响

EFFECT OF ICARITIN ON LIPOPOLYSACCHARIDE-INDUCED INFLAMMATORY RESPONSE IN PRIMARY CULTURED CORTICAL ASTROCYTES

目的探讨淫羊藿素(ICT)对脂多糖(LPS)诱导的原代皮质星形胶质细胞肿瘤坏死因子α(TNF-α)和诱导型一氧化氮合酶(iNOS)基因表达的影响及胰岛素样生长因子1受体(IGF-1R)阻断剂JB-1的阻断作用。方法常规培养原代皮质星形胶质细胞,将其分为对照组、LPS组、ICT+LPS组和JB-1+ICT+LPS组。对照组和LPS组细胞分别给予体积分数0.01二甲基亚砜(DMSO)和1 mg/L的LPS处理,其余组在有或无JB-1(1 mg/L)预处理的情况下,给予ICT(10μmol/L)预保护1 h,再加入1 mg/L的LPS共同作用6 h。应用实时荧光定量PCR检测各组细胞TNF-α和iNOS基因的表达。结果与对照组比较,LPS组TNF-α和iNOS基因表达明显上调(F=81.98、118.60,q=20.41、25.22,P<0.01);ICT预保护能明显降低由LPS诱导的TNF-α和iNOS基因表达的上调(q=7.34、13.31,P<0.01),此作用可以被IGF-1R阻断剂JB-1所阻断(q=4.65、7.52,P<0.05)。结论ICT能够抑制LPS诱导的原代皮质星形胶质细胞的炎性反应,其抗炎机制可能与IGF-1R途径的激活有关。

Objective To investigate the effect of icaritin(ICT)on the gene expression of tumor necrosis factor-α(TNF-α)and inducible nitric-oxide synthase(iNOS)in primary cultured cortical astrocytes induced by lipopolysaccharide(LPS)and the blocking effect of the insulin-like growth factor-1 receptor(IGF-1R)antagonist JB-1.Methods Primary cultured cortical astrocytes were obtained by conventional methods and were then divided into control group,LPS group,ICT+LPS group,and JB-1+ICT+LPS group.The astrocytes in the control group and the LPS group were treated with volume fraction 0.01 dimethyl sulfoxide and 1 mg/L LPS,respectively,and those in the other groups were pretreated with ICT(10μmol/L)for 1 hour with or without JB-1(1 mg/L),followed by LPS(1 mg/L)treatment for another 6 hours.Quantitative real-time PCR was used to measure the gene expression of TNF-αand iNOS in each group.Results Compared with the control group,the LPS group had significantly upregulated gene expression of TNF-αand iNOS(F=81.98,118.60;q=20.41,25.22;P<0.01).Pretreatment with ICT significantly inhibited LPS-induced upregulation of TNF-αand iNOS(q=7.34,13.31;P<0.01),which was blocked by the IGF-1R antagonist JB-1(q=4.65,7.52;P<0.05).Conclusion ICT can inhibit LPS-induced inflammatory response in primary cultured cortical astrocytes,and its anti-inflammatory mechanism may be related to activation of the IGF-1R pathway.