北京大学学报（医学版）2013，Vol.45Issue(3)：489-492,4.DOI:10.3969/j.issn.1671-167X.2013.03.029
活细胞硫化氢检测新方法
A new methods for determining hydrogen sulfide release in cultured cells
摘要
Abstract
Objective:To establish a simple method of measuring hydrogen sulfide (H2S) in cultured living cells.Methods:Filtration membrane was stuck on the lid of cell culture plate.H2S released from cultured cells was trapped by zinc acetate to generate ZnS deposition.Then the ZnS trapped in the filtration membrane was measured by methylene blue assay and the H2S production from the living cells was counted according to the standard curve.This simple method was used to access the H2S release in HepG2 (high expression CBS and CSE) and HUVEC (low expression CSE) cell lines.Results:H2S generation in cultured HepG2 cells assayed using the present method was (859.39 ± 19.12) nmol/(min · 106 cells).PAG (CSE inhibitor),HA (CBS inhibitor) or the two-inhibitor (PAG ± HA) treatment significantly lowered H2S release,respectively:(341.34 ± 105.90) nmol/(min · 106 cells),(375.05 ± 174.50) nmol/(min · 106cells),and (204.47 ± 97.14) nmol/(min · 106cells).The H2S production of HUVEC was (26.23 ± 3.24) nmol/(min · 106 cells) (about 1/30 production of HepG2 cell).Trypan blue assay showed that the cell viability was greater than 95％,suggesting that there was no cytotoxicity by using the present instrument.Conclusion:The modified instrument in cell culture plate lid was feasible for detection of hydrogen sulfide release in living cells.关键词
硫化氢/细胞/亚甲基蓝/胱硫醚β合酶/胱硫醚γ裂解酶Key words
Hydrogen sulfide/ Cells / Methylene blue/ Cystathionine gamma-lyase/ Cystathionine beta-synthase分类
医药卫生引用本文复制引用
谢静,曾强,郑扬,廖锋,徐国恒,唐朝枢,耿彬..活细胞硫化氢检测新方法[J].北京大学学报（医学版）,2013,45(3):489-492,4.基金项目
国家自然科学基金(81170235)和“十二五”国家科技支撑计划(2012BAI37B04)项目资助Supported by the National Natural Science Foundation of China (81170235) and the National Science and Technology Pillar Program during the 12th Five-Year Plan (2012BAI37B04) （81170235）
