[Monitoring early toxicity of heavy metals including Hg using a HSE-SEAP reporter gene].
Zhongguo Zhong Yao Za Zhi
; 31(16): 1346-9, 2006 Aug.
Article
de Zh
| MEDLINE
| ID: mdl-17061558
ABSTRACT
OBJECTIVE:
To develop a cellular assay based on heat shock signal pathway and secreted alkaline phosphatase (SEAP) reporter gene for investigating/predicting the early toxicity of heavy metals on HeLa cells in Chinese traditional medicine (TCM).METHOD:
The pHSE-SEAP plasmid was transfected into HeLa cells to build a HSE-SEAP-HeLa cell model. For validation of the model, the transfected cells were treated by either heating at 42 degrees C for 1 h or incubated with 5 mol x L(-1) CdCl2 for 4 h. Then the cells were covered in complete DMEM culture medium for 48 h and the activity of SEAP (reflecting the cellular level of heat shock protein) in cultural supernatants was measured; meanwhile, cell viability was determined by MTT assays. In addition, the cells were treated by four mercury compounds, HgCl2, merthilate sodium, HgS and cinnabar at the sub-lethal concentrations (determined by MTT assays). Then the heat shock response was detected likewise.RESULT:
Significant level of secreted alkaline phosphatase (SEAP) was found in pHSE-SEAP transfected HeLa cells treated either by heating (42 degrees C) or incubating with CdCl2. The heat shock protein was induced by CdCl2 before decrease of cell viability was observed. All four mercury compounds induced heat shock response in both time and concentration-dependant manner. However, there were big differences among the mercury compounds, suggesting potential differences for early-stage toxicity in vivo.CONCLUSION:
The pHSE-SEAP transfected HeLa cells respond effectively to heat shock and metal stresses, and therefore provide a practical and repeatable assay for investigating/predicting the early toxicity of heavy metals and mineral-containing drugs in TCM.
Recherche sur Google
Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
Composés du mercure
/
Gènes rapporteurs
/
Métaux lourds
/
Phosphatase alcaline
/
Protéines du choc thermique
Type d'étude:
Prognostic_studies
Limites:
Humans
Langue:
Zh
Journal:
Zhongguo Zhong Yao Za Zhi
Sujet du journal:
FARMACOLOGIA
/
TERAPIAS COMPLEMENTARES
Année:
2006
Type de document:
Article
Pays d'affiliation:
Chine