中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2011年
5期
367-371
,共5页
屈丹丹%彭福江%刘莉%杨淑玲%郭亚兵
屈丹丹%彭福江%劉莉%楊淑玲%郭亞兵
굴단단%팽복강%류리%양숙령%곽아병
肝细胞%氧化性应激%臭氧%抗氧化酶%Nrf2
肝細胞%氧化性應激%臭氧%抗氧化酶%Nrf2
간세포%양화성응격%취양%항양화매%Nrf2
Hepatocytes%Oxidative stress%Ozone%Antioxidant enzymes%Nrf2
目的 研究臭氧化盐水对肝组织细胞Keap1-核因子E2相关因子2(Nrf2)-抗氧化元件(ARE)通路中Nrf2的作用.方法 采用成年健康雄性Sprague-Dawley大鼠,随机分为正常对照组(NC组)、模型组、臭氧等渗盐水(OS)组,OS对照组(OSC组).OS组、OSC组分别予5 ml/kg OS,模型组予5 ml/kg氧气盐水每日尾静脉注射,连续15 d,第16天分别予OS组及模型组50%CCl4橄榄油溶液2ml/kg腹腔注射造肝损伤模型.NC组及OSC组予植物油2ml/kg腹腔注射,24 h后,检测大鼠血清ALT、AST、肝组织总抗氧化能力(TAOC)、还原型谷胱甘肽(GSH)、谷胱甘肽过氧化物酶(GPx)、过氧化氢酶(CAT).再提取处死大鼠肝组织的核蛋白,应用Western blot测定其细胞核中Nrf2的含量,免疫荧光组织化学技术检测细胞内Nrf2的分布.结果 与模型组比较,OS组大鼠ALT、AST降低[(1240.4±188.2)U/L、(1245.4±176.9)U/L对比(539.8±175.3)U/L、(546.0±130.2)U/L)],差异有统计学意义(P<0.01),TAOC、GSH,GPx,CAT 活性升高,分别为(0.72±0.24)U/mg、(1.05±0.21)mg/g,(676.9±115.1)U/mg、(45.2±14.3)U/mg对比(1.37±0.19)U/mg、(2.23±0.55)mg/g、(1024.6±162.9)U/mg、(68.2±9.9)U/mg,差异有统计学意义(P<0.01).与NC组比较,OSC组大鼠肝组织TAOC、GSH、GPx,CAT活性升高,差异有统计学意义(P<0.01或P<0.05).Western blot及免疫荧光均显示O3能增强肝细胞核内Nrf2的表达,Keap1-Nrf2-ARE通路的激活在O3抗氧化过程中发挥了重要的作用.结论 臭氧化盐水静脉注射可减轻CCl4所致大鼠肝损伤.其机制可能通过激活Keap1-Nrf2-ARE通路及其下游基因,增强细胞抗氧化和抗自由基的能力.
目的 研究臭氧化鹽水對肝組織細胞Keap1-覈因子E2相關因子2(Nrf2)-抗氧化元件(ARE)通路中Nrf2的作用.方法 採用成年健康雄性Sprague-Dawley大鼠,隨機分為正常對照組(NC組)、模型組、臭氧等滲鹽水(OS)組,OS對照組(OSC組).OS組、OSC組分彆予5 ml/kg OS,模型組予5 ml/kg氧氣鹽水每日尾靜脈註射,連續15 d,第16天分彆予OS組及模型組50%CCl4橄欖油溶液2ml/kg腹腔註射造肝損傷模型.NC組及OSC組予植物油2ml/kg腹腔註射,24 h後,檢測大鼠血清ALT、AST、肝組織總抗氧化能力(TAOC)、還原型穀胱甘肽(GSH)、穀胱甘肽過氧化物酶(GPx)、過氧化氫酶(CAT).再提取處死大鼠肝組織的覈蛋白,應用Western blot測定其細胞覈中Nrf2的含量,免疫熒光組織化學技術檢測細胞內Nrf2的分佈.結果 與模型組比較,OS組大鼠ALT、AST降低[(1240.4±188.2)U/L、(1245.4±176.9)U/L對比(539.8±175.3)U/L、(546.0±130.2)U/L)],差異有統計學意義(P<0.01),TAOC、GSH,GPx,CAT 活性升高,分彆為(0.72±0.24)U/mg、(1.05±0.21)mg/g,(676.9±115.1)U/mg、(45.2±14.3)U/mg對比(1.37±0.19)U/mg、(2.23±0.55)mg/g、(1024.6±162.9)U/mg、(68.2±9.9)U/mg,差異有統計學意義(P<0.01).與NC組比較,OSC組大鼠肝組織TAOC、GSH、GPx,CAT活性升高,差異有統計學意義(P<0.01或P<0.05).Western blot及免疫熒光均顯示O3能增彊肝細胞覈內Nrf2的錶達,Keap1-Nrf2-ARE通路的激活在O3抗氧化過程中髮揮瞭重要的作用.結論 臭氧化鹽水靜脈註射可減輕CCl4所緻大鼠肝損傷.其機製可能通過激活Keap1-Nrf2-ARE通路及其下遊基因,增彊細胞抗氧化和抗自由基的能力.
목적 연구취양화염수대간조직세포Keap1-핵인자E2상관인자2(Nrf2)-항양화원건(ARE)통로중Nrf2적작용.방법 채용성년건강웅성Sprague-Dawley대서,수궤분위정상대조조(NC조)、모형조、취양등삼염수(OS)조,OS대조조(OSC조).OS조、OSC조분별여5 ml/kg OS,모형조여5 ml/kg양기염수매일미정맥주사,련속15 d,제16천분별여OS조급모형조50%CCl4감람유용액2ml/kg복강주사조간손상모형.NC조급OSC조여식물유2ml/kg복강주사,24 h후,검측대서혈청ALT、AST、간조직총항양화능력(TAOC)、환원형곡광감태(GSH)、곡광감태과양화물매(GPx)、과양화경매(CAT).재제취처사대서간조직적핵단백,응용Western blot측정기세포핵중Nrf2적함량,면역형광조직화학기술검측세포내Nrf2적분포.결과 여모형조비교,OS조대서ALT、AST강저[(1240.4±188.2)U/L、(1245.4±176.9)U/L대비(539.8±175.3)U/L、(546.0±130.2)U/L)],차이유통계학의의(P<0.01),TAOC、GSH,GPx,CAT 활성승고,분별위(0.72±0.24)U/mg、(1.05±0.21)mg/g,(676.9±115.1)U/mg、(45.2±14.3)U/mg대비(1.37±0.19)U/mg、(2.23±0.55)mg/g、(1024.6±162.9)U/mg、(68.2±9.9)U/mg,차이유통계학의의(P<0.01).여NC조비교,OSC조대서간조직TAOC、GSH、GPx,CAT활성승고,차이유통계학의의(P<0.01혹P<0.05).Western blot급면역형광균현시O3능증강간세포핵내Nrf2적표체,Keap1-Nrf2-ARE통로적격활재O3항양화과정중발휘료중요적작용.결론 취양화염수정맥주사가감경CCl4소치대서간손상.기궤제가능통과격활Keap1-Nrf2-ARE통로급기하유기인,증강세포항양화화항자유기적능력.
Objective To study the effect of ozonized saline on the activation of the Keapl-Nrf2ARE signaling pathway in rat liver cells. Methods Twenty maleSprague-Dawley rats were randomly divided into ozonized saline(OS) group, model group, ozonized saline control (OSC) group and normal control (NC)group. The rats in OS group and model group were intravenously administered with OS or oxygen saline (5 ml/kg) respectively, once a day for 15 days, and then intraperitoneally injected with CCU dissolved in Oliver oil. The rats in OSC group were pretreated with OS for 15 days. The rats in NC group were fed normally for 15 days. On the 16th day, the rats in OSC group and NC group were intraperitoneally injected with Oliver oil (2 ml/kg) without CCU. After 24 hours of CCU or olive oil intraperitoneal injection, the serum levels of alanine transaminase (ALT) and aspertate aminotransferase (AST) were measured. The liver tissues were also collected for detection of total anti-oxygen capability (TAOC), glutathione (GSH), catalase (CAT), Glutathione peroxidase (GPx). Western Blot was used to detect Nrf2 and immunofluorescence staining assay to display intracelluar distribution of Nrf2. Results Compared with the rats in model group,the serum ALT and AST levels of rats in OS group were significantly lower (P < 0.01) ,which were (1240.4 ± 188.2) U/L and (1245.4 ± 176.9) U/L vs (539.8 ± 175.3) U/L and (546.0 ± 130.2) U/L, and the TAOC, CAT, GPx and GSH activity of rats in OS group were significantly higher, which were (0.72 ± 0.24) U/mg, (1.05 ±0.21) mg/g, (676.9 ± 115.1) U/mg and (45.2 ± 14.3) U/mg vs (1.37 ± 0.19) U/mg, (2.23 ± 0.55) mg/g,(1024.6 ± 162.9) U/mg and (68.2 ± 9.9) U/mg, respectively. In contrast with NC group, pretreatment of OS in OSC group elevated TAOC, CAT, GPx and GSH activity (P < 0.01 or P < 0.05). Ozonized saline can strengthen the Nrf2 expression in liver cells. Conclusions Preconditioning injection of ozonized saline can reduce rat's liver injury induced by CCl4- The ozonized saline, as a novel Nrf2 activator, can reduce the oxidative damage of radical oxygen species (ROS) and the deleterious substance by activating the KeaplNrf2-ARE signaling pathway and its downstream genes expression.
