秦皮素抑制NF-κB/COX-2信号通路改善PC12细胞损伤  

Fraxetin alleviates cell injurythrough the inhibition of NF-κB/COX-2

作　　者：江芮[1] 吕浩[1] 李申[2] 王丽莉[3] 

机构地区：[1]天津市安定医院临床三科,天津300222 [2]天津医科大学基础医学院,天津300070 [3]天津市安定医院临床八科,天津300222

出　　处：《沈阳药科大学学报》2017年第6期513-518,共6页Journal of Shenyang Pharmaceutical University

摘　　要：目的研究秦皮素(fraxetin)对脂多糖(lipopolysaccharide,LPS)致大鼠嗜铬细胞瘤细胞株(PC12细胞)损伤的作用并考察炎症反应中相关蛋白的变化。方法采用MTT法研究秦皮素对PC12细胞活力的作用,筛选合适给药浓度。试验共分为3组即对照组(Control组)、LPS模型组和秦皮素保护组(fraxetin+LPS组)。采用四甲基偶氮唑盐比色法(methyl thiazolyl tetrazolium,MTT)检测细胞活性变化;采用倒置显微镜和4′,6-diamidino-2-phenylindole dihydrochloride(DAPI)荧光核染色法观察细胞形态学变化;采用real-time PCR法检测细胞因子IL-1、IL-6、TNF-α及环氧合酶-2(cyclooxygenase-2,COX-2)mRNA水平;采用western blot法检测COX-2和NF-κB-p65蛋白表达水平。结果与模型组比较秦皮素保护组细胞存活率显著升高(P<0.01)。LPS模型组炎症因子IL-1、IL-6和TNF-αmRNA水平,NF-κB-p65和COX-2蛋白表达水平相对于对照组均明显升高(P<0.01)。与此同时秦皮素治疗组其炎症细胞因子mRNA水平,NF-κB和COX-2蛋白的表达水平均显著下调。结论秦皮素对LPS所致PC12细胞损伤有保护作用,其效应可能与抑制NF-κB/COX-2信号通路,降低炎症反应有关。Objective To investigate the protective effect and mechanism of fraxetin on lipopolysaccharide （LPS） induced PC12 cells injury, and the effect on inflammation-related proteins. Methods The PC12 cells were cultured, and the concentrations of fraxetin were explored by MTT, then the logarithmic growth phase of PC12 cells were divided into control group,LPS and fraxetin ＋ LPS groups. Cell viability was detected by MTT assay,inverted microscope and DAPI fluorescent staining were used to observe the apoptotic nuclear, real-time PCR method to detect IL-1, IL-6, TNF-α and COX-2 mRNA levels and western blotting method to detect the expression of COX-2 and NF-κB. Results The MTT results showed that 0. 1 nmol·L^-1 LPS significantly induced PC12 cells damage, compared with the model group, Fraxetin significantly increased PC12 cell viability（P 〈0.01 ）. Compared with the control group ,after the intervention of the 0.1 nmol·L^-1 LPS for 24 h, the levels of inflammatory cytokines IL-1, IL-6, TNF-α mRNA levels, NF-KB and COX-2 protein expression levels were significantly increased （ P 〈 0. 01 ） ; but the expression of inflammatory cytokines IL-1, IL-6 ,TNF-α, mRNA levels, NF-KB and COX-2 protein were reduced in fraxetin ＋ LPS group （P 〈 0.01 ）. Conclusions Fraxetin has protective effect on LPS-induced PC12 cell damage, and the effect may be related to the inhibition of NF-κB, COX-2 expression, reducing the release of inflammatory factors related.

关 键 词：秦皮素 PC12细胞 LPS 环氧化酶-2 NF-ΚB 

分 类 号：R96[医药卫生—药理学]