江芮,吕浩,李申,王丽莉
JIANG Rui,LYU Hao,LI Shen,WANG Li-li

• 1:天津市安定医院临床三科
• 2:天津医科大学基础医学院
• 3:天津市安定医院临床八科

摘要(Abstract)：

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-αand 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 PCl2 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-κB 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-κB 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(KeyWords)： 秦皮素;PC12细胞;LPS;环氧化酶-2;NF-κB
fraxetin;PC12 cells;LPS;cyclooxygenase-2;NF-κB

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 江芮,吕浩,李申,王丽莉
JIANG Rui,LYU Hao,LI Shen,WANG Li-li

DOI: 10.14066/j.cnki.cn21-1349/r.2017.06.012

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