摘要
【目的】研究不同浓度的镉对体外培养的小鼠肝细胞存活影响。【方法】采用组织块法启动肝细胞的原代培养,胰酶消化法传代培养,利用光镜观察、MTT细胞存活率检测、Hoechst33342荧光染色和DNA琼脂糖凝胶电泳等方法研究氯化镉对体外培养肝细胞存活的毒性作用。【结果】结果显示,在37℃,含20%胎牛血清的DMEM/F12培养条件下,肝脏组织原代培养启动第3天即有不规则多角形的细胞迁出,第8~10天长成细胞单层。5~160μmol/L氯化镉处理可显著降低体外培养的肝细胞的存活率,具有浓度和时间的依赖性,处理24和48 h对肝细胞的半致死浓度分别为30.3和21.0μmol/L。【结论】氯化镉处理导致肝细胞染色质的凝缩和DNA片段化,诱导细胞凋亡。
【Objective】To investigate the effect of cadmium on survival of cultured murine hepatocytes in vitro,primary murine hepatocytes were cultured with tissue block method and subcultured by trypsin digestion.【Method】Toxic effect of cadmium on cultured murine hepatocytes was examined with inverted phase microscope,MTT assay,fluorescent staining and DNA agarose gel electrophoresis.【Result】The results showed that cells migrated from tissues 3 days after the primary culture initiated in DMEM/F12（1∶ 1） medium supplemented with 20% fetal bovine serum（FBS） at 37 ℃.Hepatocytes in primary culture and subculture were polygonal morphology,proliferated to continuous monolayer within 8-10 days and propagated stably.Cadmium chloride（Cd C12） at the concentration of 5-160 μmol/L had obvious inhibitory effects on survival rate of murine hepatocytes in dose-and time-dependent manner.The half lethal concentration（IC50） at 24 and 48 hours was 30.3 and 21.0 μmol/L,respectively.【Conclusion】Treatment of Cd C12 caused chromatin condensation and fragmentation.Meanwhile,murine hepatocytes initiated apoptosis process.
作者
杜娟
张莉莉
王晶
商旋
常世民
DU Juan;ZHANG Li- li;WANG Jing;SHANG Xuan;CHANG Shi-min(Langfang Normal University, Hebei Langfang 065000, Chin)
出处
《西南农业学报》
CSCD
北大核心
2018年第4期830-834,共5页
Southwest China Journal of Agricultural Sciences
基金
河北省自然科学研究青年基金项目(C2018408059)
廊坊师范学院微生物学重点学科项目(201501)
廊坊师范学院教育教学改革项目(2015014)
廊坊师范学院应用型本科生物技术重点建设专业(201602)
关键词
镉
肝细胞
原代培养
细胞凋亡
Cadmium
Murine hepatocytes
Cell culture
Cell apoptosis
作者简介
杜娟（1983-）,女,河北衡水人,硕士,研究方向为生理学和生态学,E-mail：dujuan0203@163.com;通讯作者：王晶（1983-）,女,山东东平人,博士,研究方向为细胞毒理学和细胞免疫学,E-mail：oucflora@163.com.
