中华劳动卫生职业病杂志
中華勞動衛生職業病雜誌
중화노동위생직업병잡지
CHINESE JOURNAL OF INDUSTRIAL HYGIENE AND OCCUPATIONAL DISEASES
2012年
12期
883-887
,共5页
信丽丽%李晓海%邓华欣%旷聃%戴霞云%黄素丽%王峰%何美安%R.William Currie%邬堂春
信麗麗%李曉海%鄧華訢%曠聃%戴霞雲%黃素麗%王峰%何美安%R.William Currie%鄔堂春
신려려%리효해%산화흔%광담%대하운%황소려%왕봉%하미안%R.William Currie%오당춘
HSPA1A启动子%荧光素酶报告基因%焦炉逸散物%毒性评估
HSPA1A啟動子%熒光素酶報告基因%焦爐逸散物%毒性評估
HSPA1A계동자%형광소매보고기인%초로일산물%독성평고
HSPA1A promoter%Luciferase reporter gene%Coke oven emissions%Toxicity assessment
目的 探讨利用含HSPA1A(HSP70-1)启动子荧光素酶报告基因质粒的稳定转染HepG2/HSPA1A细胞评价焦炉逸散物毒性的可行性.方法 构建含HSPA1A启动子的pGL4.17/HSPA1A重组质粒并转染人HepG2细胞,建立稳定细胞株HepG2/HSPA1A.用不同浓度的炉底、炉侧和炉顶焦炉逸散物染毒HepG2/HSPA 1A细胞24h,分别测定细胞的相对荧光素酶活力、存活率、丙二醛(MDA)含量、Olive尾距和微核率.结果 与对照组相比,炉底焦炉逸散物染毒组细胞的相对荧光素酶活力明显升高,差异有统计学意义(P＜0.01).且在0.15 μg/L时,细胞的相对荧光素酶活力最高,为对照组的1.4倍.细胞的相对荧光素酶活力分别随着炉侧和炉顶焦炉逸散物浓度的增加而逐渐增加,差异均有统计学意义(P＜0.01).且在最高浓度时(65.4 μg/L,202 μg/L)细胞的相对荧光素酶活力分别为对照组的2.1和5.3倍.仅炉底焦炉逸散物染毒后细胞的相对荧光素酶活力与MDA浓度呈正相关,差异有统计学意义(r=0.404,P＜0.05).炉底、炉侧和炉顶焦炉逸散物染毒后,HepG2/HSPA1A细胞的相对荧光素酶活力均与Olive尾距、微核率呈正相关(r尾距分别为0.476、0.940、0.788,r微核率分别为0.580、0.649、0.432),P＜0.05.结论 HepG2/HSPA1A细胞的相对荧光素酶活力可以敏感地反映焦炉逸散物的毒性效应,该细胞可用于快速评估职业环境复合污染物的综合毒性效应.
目的 探討利用含HSPA1A(HSP70-1)啟動子熒光素酶報告基因質粒的穩定轉染HepG2/HSPA1A細胞評價焦爐逸散物毒性的可行性.方法 構建含HSPA1A啟動子的pGL4.17/HSPA1A重組質粒併轉染人HepG2細胞,建立穩定細胞株HepG2/HSPA1A.用不同濃度的爐底、爐側和爐頂焦爐逸散物染毒HepG2/HSPA 1A細胞24h,分彆測定細胞的相對熒光素酶活力、存活率、丙二醛(MDA)含量、Olive尾距和微覈率.結果 與對照組相比,爐底焦爐逸散物染毒組細胞的相對熒光素酶活力明顯升高,差異有統計學意義(P＜0.01).且在0.15 μg/L時,細胞的相對熒光素酶活力最高,為對照組的1.4倍.細胞的相對熒光素酶活力分彆隨著爐側和爐頂焦爐逸散物濃度的增加而逐漸增加,差異均有統計學意義(P＜0.01).且在最高濃度時(65.4 μg/L,202 μg/L)細胞的相對熒光素酶活力分彆為對照組的2.1和5.3倍.僅爐底焦爐逸散物染毒後細胞的相對熒光素酶活力與MDA濃度呈正相關,差異有統計學意義(r=0.404,P＜0.05).爐底、爐側和爐頂焦爐逸散物染毒後,HepG2/HSPA1A細胞的相對熒光素酶活力均與Olive尾距、微覈率呈正相關(r尾距分彆為0.476、0.940、0.788,r微覈率分彆為0.580、0.649、0.432),P＜0.05.結論 HepG2/HSPA1A細胞的相對熒光素酶活力可以敏感地反映焦爐逸散物的毒性效應,該細胞可用于快速評估職業環境複閤汙染物的綜閤毒性效應.
목적 탐토이용함HSPA1A(HSP70-1)계동자형광소매보고기인질립적은정전염HepG2/HSPA1A세포평개초로일산물독성적가행성.방법 구건함HSPA1A계동자적pGL4.17/HSPA1A중조질립병전염인HepG2세포,건립은정세포주HepG2/HSPA1A.용불동농도적로저、로측화로정초로일산물염독HepG2/HSPA 1A세포24h,분별측정세포적상대형광소매활력、존활솔、병이철(MDA)함량、Olive미거화미핵솔.결과 여대조조상비,로저초로일산물염독조세포적상대형광소매활력명현승고,차이유통계학의의(P＜0.01).차재0.15 μg/L시,세포적상대형광소매활력최고,위대조조적1.4배.세포적상대형광소매활력분별수착로측화로정초로일산물농도적증가이축점증가,차이균유통계학의의(P＜0.01).차재최고농도시(65.4 μg/L,202 μg/L)세포적상대형광소매활력분별위대조조적2.1화5.3배.부로저초로일산물염독후세포적상대형광소매활력여MDA농도정정상관,차이유통계학의의(r=0.404,P＜0.05).로저、로측화로정초로일산물염독후,HepG2/HSPA1A세포적상대형광소매활력균여Olive미거、미핵솔정정상관(r미거분별위0.476、0.940、0.788,r미핵솔분별위0.580、0.649、0.432),P＜0.05.결론 HepG2/HSPA1A세포적상대형광소매활력가이민감지반영초로일산물적독성효응,해세포가용우쾌속평고직업배경복합오염물적종합독성효응.
Objective Using the stable HSPA 1A (HSP70-1) promoter-driven luciferase reporter HepG2 cells (HepG2/HSPA1A cells) to assess the overall toxicity of coke oven emissions.Methods The stable HepG2/HSPA1A cells were treated with different concentrations of coke oven emissions (COEs) collected from the top,side,and bottom of a coke oven battery for 24 h.After the treatments,luciferase activity,cell viability,malondialdehyde (MDA) concentration,Olive tail moment,and micronuclei frequency were determined,respectively.Results The bottom COEs induced significant increases (P＜0.01) in relative luciferase activity up to 1.4 times the control level at 0.15 μg/L.The low dose of side COEs (0.02 μg/L) led to a significant increase (P＜0.01) in relative luciferase activity that progressively increased to 2.1 times the control level at 65.4 μg/L.The top COEs produced a strong dose-dependent induction of relative luciferase activity up to over 5 times the control level at the highest concentration tested (202 μg/L).In HepG2/HSPA1A cells treated with the bottom COEs,relative luciferase activity was positively correlated with MDA concentration (r=0.404,P＜0.05).For the three COEs samples,positive correlations were observed between relative luciferase activity and Olive tail moment and micronuclei frequency.Conclusion The relative luciferase activity in HepG2/HSPA1A cells can sensitively reflect the overall toxicity of COEs.The stable HepG2/HSPA1A cells can be used for rapid screening of the overall toxicity of complex air pollutants in the workplace.
