摘要
目的探讨不同条件下诱导L02肝细胞损伤的最佳条件,建立稳定可靠的体外肝细胞损伤模型。方法将常规培养的L02细胞分别设置为正常对照组、过氧化氢(H_(2)O_(2))组、对乙酰氨基酚(APAP)组和乙醇组;H_(2)O_(2)组用不同浓度梯度(300、400、500和600 mol/L)的H_(2)O_(2)分别诱导6、12和16 h;APAP组以不同浓度梯度(1、10、20和40 mmol/L)分别诱导3和6 h;乙醇组加入不同浓度梯度(0.5%、1%、1.5%和2%)的乙醇分别孵育6和24 h;在上述损伤条件下用细胞计数试剂盒(CCK)-8检测各组L02细胞的存活率。在确定的最佳损伤条件下,建立不同肝损伤模型,检测丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、丙二醛(MDA)和超氧化物歧化酶(SOD)等损伤指标。结果除300 mol/L H_(2)O_(2)诱导12 h以外,其余不同浓度H_(2)O_(2)组的细胞存活率均低于正常对照组(均<0.05)。不同浓度APAP组的细胞存活率均低于正常对照组(均<0.05),尤其是40 mmol/L的APAP更为明显。乙醇诱导6 h后,不同浓度乙醇组的细胞存活率均明显下降,1.5%和2%乙醇的细胞存活率均低于正常对照组(均<0.05);此外,不同浓度的乙醇诱导24 h可以引起L02细胞大量坏死。H_(2)O_(2)模型组、APAP模型组和乙醇模型组的细胞存活率、SOD活性均低于正常对照组(均<0.05),而ALT、AST、MDA含量均高于正常对照组(均<0.05)。结论分别用300mol/L H_(2)O_(2)诱导6 h、40 mmol/L APAP作用3 h和1.5%的乙醇作用6 h的L02肝细胞所形成的肝损伤模型比较稳定,是体外模拟氧化性、药物性和酒精性肝损伤模型的最佳诱导条件。
Objective To study the optimum conditions of the L02 hepatocytes injured under different conditions,and establish a stable and reliable model of hepatocytes injured in vitro.Methods The L02 hepatocytes were divided into the normal control group,hydrogen peroxide(H_(2)O_(2))group,Acetaminophen(APAP)group andethanol group.In the H_(2)O_(2)group,the L02 hepatocytes were treated with different concentration gradient of H_(2)O_(2)(300,400,500 and 600 mol/L)for 3,12 and 16 h,respectively.In the APAP group,APAP with different concentration gradients(1,10,20 and 40 mmol/L)was induced for 3 and 6 h,respectively.In the ethanol group,ethanol with different concentration gradients(0.5%,1%,1.5%and 2%)was induced for 6 h and 24 h,respectively.Cell viability was tested with cell coun-ting kit(CCK)-8 among all the groups.Under the optimal injury conditions,different models of liver injury were es-tablished,and the indexes of liver injury such as malondialdehyde(MDA)and superoxide dismutase(SOD)were de-tected,and the stability of the models was verified.Results Compared to the normal control group,the other con-centration of H_(2)O_(2)had significant differences except 300 mol/L of H_(2)O_(2)induced for 12 h(all<0.05).Cell viability declined with increased APAP concentration and treatment time,especially with APAP at the concentration of 40 mmol/L(<0.05).Cell viability was obviously decreased in each concentration after 6 h ethanol treatment,espe-cially with ethanol at the concentration of 1.5%and 2%(all<0.05).In addition,a large percentage of L02 hepa-tocytes were died after 24 h treatment with different concentration gradient of ethanol.The cell survival rate and SOD activity of H_(2)O_(2)group,APAP group and ethanol group were lower than those of the normal control group(all<0.05),while the contents of alanine aminotransferase(ALT),aspartate aminotransferase(AST)and MDA were higher than those of the normal control group(all<0.05).Conclusions The L02 hepatocytes separately treating with 300 mol/L of H_(2)O_(2)for 6 h,40 mmol/L
作者
姜晓杰
潘金明
琚立萍
张晶
孙晶晶
庄思静
席建军
庄让笑
JIANG Xiaojie;PAN Jinmin;JU Liping;ZHANG Jing;SUN Jingjing;ZHUANG Sijing;XI Jianjun;ZHUANG Rangxiao(Xixi Hospital of Hangzhou,Hangzhou 310023,Zhejiang,China)
出处
《现代实用医学》
2024年第2期154-158,共5页
Modern Practical Medicine
基金
浙江省基础公益研究计划项目(LGF21H300002)。
