5-羟色胺通过上调2B受体抑制放线菌素D诱导肝细胞凋亡初步实验研究  

5-HT inhibits actinomycin D induced hepatocyte apoptosis by upregulating 2B receptor

作　　者：马丽霞[1] 高玉娟 刘晓慧[1] 张晶[1] MA Li-xia;GAO Yu-juan;LIU Xiao-hui;ZHANG Jing(Department of Hepatitis C and Toxic Hepatology,Beijing Youan Hospital,Capital Medical University,Beijing 100069,China)

机构地区：[1]首都医科大学附属北京佑安医院丙型肝炎与中毒性肝病科,100069 [2]南京大学附属南京鼓楼医院呼吸与危重症医学科

出　　处：《肝脏》2019年第12期1387-1392,共6页Chinese Hepatology

摘　　要：目的探讨5-羟色胺(5-HT)在肝细胞凋亡中的作用及机制。方法以不同浓度梯度放线菌素D(AcD)分别诱导HepG2和7702细胞凋亡,采用AnnexinV-FITC/PI流式细胞术和TUNEL方法检测细胞凋亡,MTT法检测细胞存活率。AcD预处理HepG2细胞后分别加入5-HT、5-HT 2A受体激动剂DOI、5-HT 2B受体激动剂M110、5-HT 2B受体拮抗剂SB204+5-HT,AnnexinV-FITC/PI流式细胞术和MTT法分别检测细胞凋亡率和存活率,蛋白质印迹法检测蛋白水解酶3(caspase3)、丝氨酸/苏氨酸蛋白激酶(AKT)及磷酸化丝氨酸/苏氨酸蛋白激酶(p-AKT)的表达。结果AcD以浓度依赖性方式分别诱导HepG2细胞和7702细胞的凋亡,50 ng/mL AcD处理两种细胞凋亡率均高于无血清对照组(HePG2细胞t=1.71,7702细胞t=1.98,均P<0.05),与单用AcD组相比,5-HT使HepG2细胞凋亡率从31.96%±2.13%降至10.24%±2.59%(t=1.62,P<0.05),5-HT 2B受体激动剂使其降至8.41%±0.96%(t=1.75,P<0.05),反之,5-HT 2B受体拮抗剂能拮抗5-HT的抗凋亡作用,其凋亡率为25.92%±1.33%(t=1.45,P>0.05)。5-HT及5-HT 2B受体激动剂均能够减少caspase3活化,增加AKT的磷酸化,使细胞凋亡减少。结论5-HT通过上调5-HT 2B受体促进AKT磷酸化,从而抑制AcD诱导的肝细胞凋亡。Objective To investigate the role and mechanism of 5-hydroxytryptamine(5-HT)in hepatocyte apoptosis,and to provide evidence for the application of 5-HT in liver diseases.Methods Apoptosis of HepG2 and 7702 cells was induced by different concentrations of actinomycin-D(AcD).After AcD pretreatment of HepG2 cells,5-HT,5-HT 2A receptor agonist DOI,5-HT 2B receptor agonist M110,and 5-HT 2B receptor antagonist SB204 with 5-HT were added respectively.Then,apoptotic rates and survival rates were detected by Annexin V-FITC/PI flow cytometry and MTT assay,expressions of aspartate specific protease 3(caspase-3),serine/threonine protein kinase(AKT)and phosphorylated serine/threonine protein kinase(p-AKT)were detected by Western blot method.Results AcD induced apoptosis of HepG2 cells and 7702 cells in a concentration-dependent manner.The apoptotic rates of cells treated with 50 ng/ml AcD were higher than those of serum-free control group(HepG2 cells t=1.71,7702 cells t=1.98,P<0.05).Compared with the AcD group,5-HT decreased the apoptosis rate of HepG2 cells from 31.96%±2.13%to 10.24%±2.59%(t=1.62,P<0.05),and the 5-HT 2B receptor agonist decreased the apoptosis rate to 8.41%±0.96%(t=1.75,P<0.05).On the contrary,the 5-HT 2B receptor antagonist antagonize the anti-apoptotic effect of 5-HT,making the apoptotic rate 25.92%±1.33%(t=1.45,P>0.05).Both 5-HT and 5-HT 2B receptor agonists reduced the activation of caspase3,increased the phosphorylation of AKT,and decreased the apoptosis.Conclusion 5-HT promotes AKT phosphorylation by up-regulating 5-HT 2B receptor,thereby inhibiting AcD-induced hepatocyte apoptosis.

关 键 词：放线菌素D 5-HT 肝细胞凋亡 AKT磷酸化 5-HT 2B受体 

分 类 号：R73[医药卫生—肿瘤]