中华传染病杂志
中華傳染病雜誌
중화전염병잡지
CHINESE JOURNAL OF INFECTIOUS DISEASES
2012年
7期
402-406
,共5页
杨铂%张立新%刘靓雯%安勇
楊鉑%張立新%劉靚雯%安勇
양박%장립신%류정문%안용
肝炎,中毒性%细胞凋亡%异烟肼%HepG2细胞%细胞,培养的%基因表达%膜糖蛋白类%Fas/FasL
肝炎,中毒性%細胞凋亡%異煙肼%HepG2細胞%細胞,培養的%基因錶達%膜糖蛋白類%Fas/FasL
간염,중독성%세포조망%이연정%HepG2세포%세포,배양적%기인표체%막당단백류%Fas/FasL
Hepatitis,toxic%Apoptosia%Isoniazid%HepG2 cells%Cells,cultured%Gene expression%Membrane glycoproteins%Fas/FasL
目的 通过建立异烟肼致HepG2细胞坏死或凋亡模型,观察HepG2细胞Fas/Fas配体(FasL)的表达.方法 以HepG2细胞为模型,分别用含1、2、4、6、8 mg/mL异烟肼的细胞培养液,空白对照组加入新鲜培养液,培养24 h后观察各组细胞形态,膜联蛋白(Annexin)V和碘化丙啶染色,流式细胞仪检测HepG2细胞的坏死和凋亡情况以及其Fas/FasL的表达.数据采用单因素方差分析,各不同浓度药物组与空白对照组的比较采用Dunnett t检验.结果 随异烟肼浓度的增加(4、6、8 mg/mL),HepG2细胞出现逐渐增多的坏死和凋亡,总死亡率分别为(32.1±7.5)％、(34.9±8.1)％和(38.2±9.4)％,与正常对照组的(7.2±1.5)％相比,差异有统计学意义(t=4.62、5.14、5.75,均P＜0.01)；Fas的表达也随之增加,异烟肼2、4、6和8 mg/mL浓度组Fas表达率分别为(8.7±2.2)％、(11.5±2.8)％、(12.3±3.0)％和(10.6±2.9)％,与正常对照组的(3.1±0.8)％比较,差异有统计学意义(t=2.97,P＜0.05;t=4.46,P＜0.01；t=4.88,P＜0.01；t=3.98,P＜0.05).异烟肼4、6、8 mg/mL浓度组FasL表达率分别为(16.2±3.5)％、(21.7±4.8)％、(18.7±4.9)％,与正常对照组的(7.4±1.4)％相比,差异有统计学意义(t=3.11,P＜0.01;t=5.06,P＜0.01；t=3.99,P＜0.05).异烟肼浓度为8 mg/mL时,HepG2细胞的死亡增加,主要以坏死为主,凋亡发生率未再增加.结论 异烟肼可以诱导HepG2细胞变性、坏死和凋亡,这种凋亡的发生可能与异烟肼诱导肝细胞表达Fas/FasL增多有关.
目的 通過建立異煙肼緻HepG2細胞壞死或凋亡模型,觀察HepG2細胞Fas/Fas配體(FasL)的錶達.方法 以HepG2細胞為模型,分彆用含1、2、4、6、8 mg/mL異煙肼的細胞培養液,空白對照組加入新鮮培養液,培養24 h後觀察各組細胞形態,膜聯蛋白(Annexin)V和碘化丙啶染色,流式細胞儀檢測HepG2細胞的壞死和凋亡情況以及其Fas/FasL的錶達.數據採用單因素方差分析,各不同濃度藥物組與空白對照組的比較採用Dunnett t檢驗.結果 隨異煙肼濃度的增加(4、6、8 mg/mL),HepG2細胞齣現逐漸增多的壞死和凋亡,總死亡率分彆為(32.1±7.5)％、(34.9±8.1)％和(38.2±9.4)％,與正常對照組的(7.2±1.5)％相比,差異有統計學意義(t=4.62、5.14、5.75,均P＜0.01)；Fas的錶達也隨之增加,異煙肼2、4、6和8 mg/mL濃度組Fas錶達率分彆為(8.7±2.2)％、(11.5±2.8)％、(12.3±3.0)％和(10.6±2.9)％,與正常對照組的(3.1±0.8)％比較,差異有統計學意義(t=2.97,P＜0.05;t=4.46,P＜0.01；t=4.88,P＜0.01；t=3.98,P＜0.05).異煙肼4、6、8 mg/mL濃度組FasL錶達率分彆為(16.2±3.5)％、(21.7±4.8)％、(18.7±4.9)％,與正常對照組的(7.4±1.4)％相比,差異有統計學意義(t=3.11,P＜0.01;t=5.06,P＜0.01；t=3.99,P＜0.05).異煙肼濃度為8 mg/mL時,HepG2細胞的死亡增加,主要以壞死為主,凋亡髮生率未再增加.結論 異煙肼可以誘導HepG2細胞變性、壞死和凋亡,這種凋亡的髮生可能與異煙肼誘導肝細胞錶達Fas/FasL增多有關.
목적 통과건립이연정치HepG2세포배사혹조망모형,관찰HepG2세포Fas/Fas배체(FasL)적표체.방법 이HepG2세포위모형,분별용함1、2、4、6、8 mg/mL이연정적세포배양액,공백대조조가입신선배양액,배양24 h후관찰각조세포형태,막련단백(Annexin)V화전화병정염색,류식세포의검측HepG2세포적배사화조망정황이급기Fas/FasL적표체.수거채용단인소방차분석,각불동농도약물조여공백대조조적비교채용Dunnett t검험.결과 수이연정농도적증가(4、6、8 mg/mL),HepG2세포출현축점증다적배사화조망,총사망솔분별위(32.1±7.5)％、(34.9±8.1)％화(38.2±9.4)％,여정상대조조적(7.2±1.5)％상비,차이유통계학의의(t=4.62、5.14、5.75,균P＜0.01)；Fas적표체야수지증가,이연정2、4、6화8 mg/mL농도조Fas표체솔분별위(8.7±2.2)％、(11.5±2.8)％、(12.3±3.0)％화(10.6±2.9)％,여정상대조조적(3.1±0.8)％비교,차이유통계학의의(t=2.97,P＜0.05;t=4.46,P＜0.01；t=4.88,P＜0.01；t=3.98,P＜0.05).이연정4、6、8 mg/mL농도조FasL표체솔분별위(16.2±3.5)％、(21.7±4.8)％、(18.7±4.9)％,여정상대조조적(7.4±1.4)％상비,차이유통계학의의(t=3.11,P＜0.01;t=5.06,P＜0.01；t=3.99,P＜0.05).이연정농도위8 mg/mL시,HepG2세포적사망증가,주요이배사위주,조망발생솔미재증가.결론 이연정가이유도HepG2세포변성、배사화조망,저충조망적발생가능여이연정유도간세포표체Fas/FasL증다유관.
Objective To establish a model of isoniazid induced necrosis and apoptosis in HepG2 cell and to observe the expressions of Fas/Fas ligand (FasL) in this model.Methods HepG2 cells were treated with different dosages of isoniazid (0,1,2,4,6 and 8 mg/mL) or blank control for 24 hours.Flow cytometer was used to observe the cellular morphology of the HepG2 cell.Annexin V/propidium iodide double staining and flow cytometry were employed to detect the necrosis and apoptosis of HepG2 cells.The expressions of Fas/FasL on the cells were also determined by flow cytometry.The data were analyzed by one-way ANOVA.The comparisons between the drug groups and the control group were performed by using Dunnett t test. Results The higher the dose of isoniazid (4,6,8 mg/mL) was,the more necrosis and apoptosis were observed.In the 4,6 and 8mg/mL isoniazid arms,the total mortality rates were all higher than the control group [(32.1 ±7.5)％,(34.9±8.1)％,(38.2±9.4)％ vs (7.2±1.5)％ respectively](t=4.62,5.14 and 5.75,respectively; all P＜0.01 ).The expression levels of Fas increased along with the dose of isoniazid increasing [(8.7±2.2)％,(11.5±2.8)％,(12.3±3.0)％ and (10.6±2.9)％ in isoniazid 2,4,6and 8 mg/mL arms,respectively],which were all higher than that in control arm [(3.1 ±0.8) ％](t=2.97,P＜0.05; t=4.46,P＜0.01; t=4.88,P＜0.01; t=3.98,P＜0.05).Furthermore,the expressions of FasL increased as well when the dose of isoniazid increased.The expression levels of FasL were (16.2±3.5)％,(21.7±4.8)％ and (18.7±4.9)％,respectively in isoniazid 4,6 and 8 mg/mL arms,which were all higher than that in the control group [(7.4±1.4)％](t=3.11,P＜0.01; t=5.06,P＜0.01; t=3.99,P＜0.05).HepG2 cell necrosis increased with isoniazid of 8 mg/mL.However,the increase of apoptosis was not observed.Conclusion Isoniazid can induce HepG2 cell necrosis and apoptosis,and the apoptosis may be related with the increased expressions of the Fas/FasL on the cells.
