本文選題：硫化氫　切入點(diǎn)：細(xì)胞　出處：《北京大學(xué)學(xué)報(bào)(醫(yī)學(xué)版)》2013年03期

【摘要】：目的:建立測定活細(xì)胞釋放微量硫化氫(hydrogen sulfide,H2S)的簡易方法。方法:在細(xì)胞培養(yǎng)板蓋上方黏附濾膜,細(xì)胞釋放的硫化氫與濾膜上的醋酸鋅反應(yīng)產(chǎn)生硫化鋅,采用亞甲基藍(lán)分光光度法測定并計(jì)算細(xì)胞釋放的硫化氫的量。應(yīng)用該方法分別檢測了HepG2細(xì)胞和人臍靜脈內(nèi)皮細(xì)胞硫化氫的釋放量。結(jié)果:HepG2細(xì)胞添加L-半胱氨酸以及輔酶磷酸吡哆醛后,繼續(xù)培養(yǎng)12 h,H2S釋放量為(859.39±19.12)nmol/(min.106cells);給予胱硫醚-γ-裂解酶抑制劑炔丙基甘氨酸(DL-propargylglycine,PAG)后,H2S產(chǎn)率下降到(341.34±105.90)nmol/(min.106cells);胱硫醚-β-合酶抑制劑間羥胺處理后,H2S產(chǎn)率下降到(375.05±174.50)nmol/(min.106cells);兩種酶抑制劑聯(lián)合使用后H2S釋放量顯著抑制,為(204.47±97.14)nmol/(min.106cells)。人臍靜脈內(nèi)皮細(xì)胞也可內(nèi)源性產(chǎn)生H2S,HUVEC細(xì)胞H2S的釋放量為(26.23±3.24)nmol/(min.106cells),約為HepG2細(xì)胞產(chǎn)量的1/30。臺(tái)盼藍(lán)檢測細(xì)胞活性大于95%,細(xì)胞生長狀態(tài)良好,表明該方法無細(xì)胞毒作用,細(xì)胞可根據(jù)實(shí)驗(yàn)需要繼續(xù)培養(yǎng)。結(jié)論:濾膜吸附法簡便易行、實(shí)用可靠,可應(yīng)用于活細(xì)胞釋放硫化氫的測定。
[Abstract]:Objective: to establish a simple method for the determination of trace hydrogen sulfide (H _ 2S) released from living cells.Methods: the membrane was adhered to above the cell culture plate. The hydrogen sulfide released by the cell reacted with zinc acetate on the filter membrane to produce zinc sulfide. The amount of hydrogen sulfide released by the cell was determined and calculated by methylene blue spectrophotometry.The release of hydrogen sulfide from HepG2 cells and human umbilical vein endothelial cells (HUVEC) was measured by this method.Results after addition of L- cysteine and coenzyme pyridoxal,緇х畫鍩瑰吇12 h,H2S閲婃斁閲忎負(fù)(859.39鹵19.12)nmol/(min.106cells);緇欎簣鑳辯～閱，

本文編號：1711463