氟西汀对PC12细胞缺氧损伤的保护作用  

Protective effect of fluoxetine against hypoxia induced injury on PC12 cells

作　　者：曹琪璐 刘菁 赵彤 陈克明 马慧萍[1,2] 贾正平 CAO Qi-lu;LIU Jing;ZHAO Tong;CHEN Ke-ming;MA Hui-ping;JIA Zheng-ping(School of Pharmacy,Lanzhou University,Lanzhou 730000,China;PLA Joint Logistic Support Hospital 940 Hospital,Lanzhou 730050,China)

机构地区：[1]兰州大学药学院,甘肃兰州730000 [2]中国人民解放军联勤保障部队第九四〇医院,甘肃兰州730050

出　　处：《中国药理学通报》2021年第6期866-870,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金面上项目(No 81571847);军队后勤科研计划(No CWH17J010)。

摘　　要：目的以PC12细胞为研究对象,探讨氟西汀对体外培养细胞缺氧损伤的保护作用。方法将PC12细胞随机分为常氧对照组、缺氧模型组和缺氧+盐酸氟西汀组(盐酸氟西汀组)。将后两组一同放入缺氧培养盒中培养18 h后,观察细胞状态,CCK-8法检测细胞活力,DCFH-DA荧光染色法检测活性氧(ROS)含量、酶标仪法检测丙二醛(MDA)水平和乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性;Western blot法检测凋亡蛋白Bax、Bcl-2和caspase-3等的表达。结果与常氧对照组相比,缺氧导致PC12细胞出现明显损伤,盐酸氟西汀在10-6 mol·L-1对PC12细胞缺氧损伤有最大保护作用;与正常对照组相比,缺氧导致PC12细胞LDH、ROS和MDA含量显著升高,SOD和CAT活性降低,盐酸氟西汀处理能逆转这些变化;盐酸氟西汀还能够下调Bax和caspase-3蛋白的表达,上调Bcl-2的表达。结论盐酸氟西汀对缺氧致PC12细胞损伤具有保护作用,其作用机制可能与缓解氧化应激、抑制细胞凋亡有关。Aim To study the protective effect of fluoxetine against hypoxia induced injury on PC12 cells.Methods PC12 cells were randomly divided into control group,hypoxia group,and fluoxetine hydrochloride group.The last two groups were put into a hypoxic culture chamber for 18 hours,the cell state was observed under inverted microscope,and cell viability was detected using CCK-8 assay.Reactive oxygen species(ROS)level was evaluated by DCFH-DA.Lactate dehydrogenase(LDH),superoxide dismutase(SOD),catalase(CAT),malondialdehyde(MDA)in cell culture supernatant were evaluated by enzyme labeling method.The expression levels of Bcl-2,Bax and caspase-3 were determined by Western blot.Results Compared with normal group,hypoxia caused obvious damage to PC12 cells.Fluoxetine hydrochloride at 10-6 mol·L-1 had a significant protective effect on PC12 cells against hypoxia damage,the viability as well as the activity of antioxidant enzyme such as SOD and CAT was reduced and the level of LDH,MDA and ROS increased when treated with hypoxia in PC12 cells.These changes were reversed with the pretreatment of fluoxetine.In addition,fluoxetine could inhibit the expression of Bax,caspase-3,while upregulate the expression of Bcl-2 expression.Conclusions Fluoxetine can suppress the hypoxia-induced damage on PC12 cells,and the mechanism may be related to scavenging free radicals,increasing the activity of antioxidant enzymes decreasing oxidative stress,and inhibiting inflammatory response and apoptosis.

关 键 词：氟西汀 缺氧 PC12 活性氧 氧化应激 凋亡蛋白 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]