间苯三酚对肝缺血再灌注损伤的作用及机制研究  

Role and mechanism of phloroglucinol in hepatic ischemia reperfusion injury

作　　者：吴建波[1] 杨建纲[1] 殷莉波[1] 倪沂江[1] 周照[1] 程宇[1] 李圆[1] 

机构地区：[1]南京医科大学附属常州第二人民医院普外科,江苏常州213000

出　　处：《南京医科大学学报（自然科学版）》2015年第2期196-200,共5页Journal of Nanjing Medical University(Natural Sciences)

摘　　要：目的 :研究间苯三酚(phloroglucinol,Phlo)在肝脏缺血再灌注损伤(ischemia reperfusion injury,IRI)中的作用。方法 :将40只C57BL/6小鼠随机分为假手术(sham)组、IR组、IR+Phlo组、IR+磷酸缓冲盐(phosphate buffer saline,PBS)组4组,每组10只。Sham组仅分离暴露门静脉,不夹闭。另3组建立小鼠部分肝缺血再灌注模型(70%)。利用小鼠正常巨噬细胞系RAW264.7,进行体外脂多糖(lipopolysaccharide,LPS)刺激,建立体外模型。通过血清丙氨酸转氨酶(alanine transaminase,ALT)、天门冬氨酸转氨酶(aspartate transaminase,AST)的水平和组织病理学评价肝脏损伤,利用实时荧光定量PCR检测肝内IL-1β和TNF-α的m RNA水平变化,细胞培养上清液IL-1β和TNF-α的水平由ELISA检测,RAW264.7细胞内p-p65蛋白表达情况经Western blot进行测定。结果 :IR+Phlo组小鼠肝损伤情况明显低于IR组与IR+PBS组(P<0.01),肝内IL-1β和TNF-α的m RNA水平均降低(P<0.05)。体外试验证实间苯三酚与RAW264.7共培养时,可减少其分泌的IL-1β和TNF-α(P<0.05),细胞中NF-κB的活性也明显下降。结论:间苯三酚在小鼠肝脏缺血再灌注损伤中具有保护作用,该作用部分是通过间苯三酚抑制巨噬细胞内NF-κB的活性实现。Objective：To investigate the role and mechanism of phloroglucinol in hepatic ischemia reperihsion injury. Methods： Forty C57BL/6 nlice were randomly divided into four groups on average, named as sham group,IR group,IR＋Phlo group, and 1R＋PBS group. The sham group only separated portal vein without clipping,while partial liver ischemia reperfusion model（70%）was estab- lished in other three groups. RAW264.7,a mouse normal macrophage cell line,was incubated with or without phloroglucinol followed by stimulation of LPS in vitro. We evaluated the degree of liver injury by examining the levels of serum ALT and AST and histopathology graded by Suzuki＇s score. Expressions of liver IL-113 and TNF-o~ mRNA were detected in real-time qPCR. ELISA was used to measure the levels of supernatant 1L-1 [3 and TNF-~ of RAW264.7. The expression of phospho-p65 in RAW264.7 was assessed by western blot. Results：The degree of liver injury and the levels of liver IL-1 [3 and TNF-~ mRNA in IR＋Phlo group decreased compared to those in IR and IR＋PBS groups（P 〈 0.05）. Preineubation with phloroglucinol could significantly inhibit the ablity of secretion of [L-I [3 and TNF-~ by RAW264.7 （P 〈 0.05）,which involved the supression of the activation of NF-KB. Conclusion：Phloroglucinol plays a protective role during hepatic ischemia reperfusion injury,which is partially relates to the supression of the activation of NF-KB in macrophage.

关 键 词：肝缺血再灌注损伤 间苯三酚 RAW264.7 NF-ΚB 

分 类 号：R757.3[医药卫生—皮肤病学与性病学]