银杏叶提取物对NF-κB活性抑制人视网膜色素上皮细胞炎症反应的影响  

Inflammation Inhibitory Effect of Ginkgo Leaves Extract on NF-κB Activity in Human Retinal Pigment Epithelial Cells

作　　者：南娜[1] 赵萍[1] 张睿[2] 

机构地区：[1]河北医科大学第四医院,石家庄050011 [2]河北医科大学第一医院,石家庄050031

出　　处：《中国实验方剂学杂志》2017年第18期163-167,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81400812)

摘　　要：目的:观察银杏叶提取物对高糖诱导的人视网膜色素上皮细胞ARPE-19炎症的影响及核转录因子-kappaB(NF-κB)通路的变化,探讨其作用机制。方法:体外培养ARPE-19细胞株,分为正常组(5.5 mmol·L-1葡萄糖),高糖组(30 mmol·L-1葡萄糖),银杏叶提取物不同剂量组组(30 mmol·L-1葡萄糖+12.5,25,50,100 mg·L-1银杏叶提取物)。噻唑蓝(MTT)比色法检测细胞活性;流式细胞术测定细胞中肿瘤坏死因子-α(TNF-α),白细胞介素-1β(IL-1β)的含量;实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白质免疫印迹法(Western blot)测定炎症因子(NIK/p-NIK,IKKα/p-IKKα,IκB/p-IκB)表达;Western blot检测NF-κB磷酸化及核NF-κB表达。结果:与正常组比较,高糖组吸光度A明显降低(P<0.05),细胞TNF-α,IL-1β含量明显升高(P<0.05),p-NIK,p-IKKα,p-IκB表达水平明显升高(P<0.05),p-NF-κB,核NF-κB表达水平明显升高(P<0.05);与高糖组比较,12.5,25,50,100 mg·L-1银杏叶提取物组吸光度A明显升高(P<0.05);细胞TNF-α,IL-1β含量明显降低(P<0.05),p-NIK,p-IKKα,p-IκB水平明显降低(P<0.05),p-NF-κB,核NF-κB表达水平明显降低(P<0.05)。结论:银杏叶提取物能通过抑制NF-κB活性抑制炎症反应,保护视网膜色素上皮细胞,用于糖尿病视网膜病变的防治。Objective： To observe the effect of ginkgo leaves extract on the inflammation of humanesophageal epithelial cell ARPE-19 line and the changes in nuclear factor-kappa B（ NF-κB） pathway,in order to explore its mechanism. Method： In vitro,ARPE-19 cells was divided into normal control group（ 5. 5 mmol·L-1glucose）,high-glucose group（ 30 mmol·L-1glucose） and ginkgo leaves extract group（ 30 mmol·L-1glucose ＋12. 5,25,50,100 mg·L^（-1）ginkgo leaves extract）. MTT assay was used to detect cell viability. The levels of tumor necrosis factor-α（ TNF-α） and interleukin-1β（ IL-1β） in the cells were determined by flow cytometry. NIK/pNIK,IKKα/p-IKKα and IκB/p-IκB expressions were measured by Real-time PCR and Western blot. Western blot was used to detect the phosphorylation of NF-κB and the expression of nuclear NF-κB. Result： The A value of the high-glucose group was significantly lower than that of the control group,ginkgo leaves extract groups（ 12. 5,25,50,100 mg·L^（-1）） were significantly higher than those of the high-glucose group（ P〈0. 05）. Levels of TNF-α and IL-1β of the high-glucose group was significantly higher than that of the control group,and those of ginkgo leaves extract group（ 12. 5,25,50,100 mg·L^（-1）） were significantly lower than those of high glucose group（ P〈0. 05）.Compared with the control group,the expressions of p-NIK,p-IKKα,p-IκB in the high-glucose group increased,and the expressions of p-NIK,p-IKKα,p-IκB were decreased in ginkgo leaves extract（ 12. 5,25,50 and 100 mg·L^（-1）） groups（ P〈0. 05）. The phosphorylation and nuclear NF-κB protein expression were increased in the highglucose group compared with the control group,and the NF-κB expression were decreased in ginkgo leaves extract groups（ 12. 5,25,50,100 mg·L^（-1）,P〈0. 05）. Conclusion： ginkgo leaves extract can inhibit the inflammatory response by inhibiting NF-κB activity and protect retinal pigment epithelial cells,so as to prevent a

关 键 词：银杏叶提取物 高糖 糖尿病视网膜病变 炎症反应 核转录因子kappaB活性 

分 类 号：R587[医药卫生—内分泌] R285.5[医药卫生—内科学]