PQ401的体内外抗炎效应研究

发布时间：2018-08-13 10:50

【摘要】：目的研究胰岛素样生长因子-1受体(insulin-like growth factor 1 receptor,IGF-1R)的选择性抑制剂PQ401在体内外的抗炎效应。方法体外实验:小鼠RAW264.7细胞株经0.1、1、10μmol/L PQ401预作用1 h后,加入10μg/m L脂多糖(LPS)作用12 h,采用酶联免疫吸附实验(ELISA)检测细胞上清肿瘤坏死因子-α(TNF-α)、白细胞介素(IL-1β、IL-6)的含量。另将小鼠RAW264.7细胞株经10μmol/L PQ401预作用1 h后,加入10μg/m L LPS作用6、12、24 h后再次检测以上指标的变化。体内实验:将50只成年雄性C57BL/6小鼠按随机数字表法分为对照组、模型组和不同剂量PQ401组,每组10只。腹腔注射LPS 10 mg/kg制备急性炎症反应小鼠模型,对照组腹腔注射等量二甲基亚砜(DMSO),PQ401组分别腹腔注射LPS 10 mg/kg和PQ401 25、50、100 mg/kg。于给药12 h后通过ELISA检测血清和肝组织中TNF-α、IL-1β、IL-6的含量;用全自动生化分析仪测定血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)及总胆红素(TBil)水平;采用Western blot检测肝细胞核内核转录因子-κB p65(NF-κB p65)的含量。结果与LPS单独作用组相比,PQ401呈浓度和时间依赖性降低RAW264.7细胞上清中TNF-α、IL-1β、IL-6的水平。在LPS诱导的小鼠急性腹膜炎模型中,小鼠经LPS注射12 h后表现为精神萎靡、活动减少,而PQ401组小鼠的精神状态、活动均好于模型组。PQ401治疗后能有效降低LPS所致急性腹膜炎小鼠血清中TNF-α、IL-1β、IL-6的含量,降低肝组织TNF-α、IL-1β的水平和NF-κB p65的含量。结论 PQ401在体内外对LPS诱导的炎症因子分泌具有明显的抑制效应。
[Abstract]:Objective to study the anti-inflammatory effect of PQ401, a selective inhibitor of insulin-like growth factor-1 receptor (insulin-like growth factor 1 receptor) in vitro and in vivo. Methods: mouse RAW264.7 cell line was pretreated with 0.1 渭 mol/L PQ401 for 1 h, then 10 渭 g / mL lipopolysaccharide (LPS) was added for 12 h. The levels of tumor necrosis factor 伪 (TNF- 伪) and interleukin-6 (IL-1 尾 -IL-6) in the supernatant were measured by Elisa (ELISA). In addition, the mouse RAW264.7 cell line was pretreated with 10 渭 mol/L PQ401 for 1 h, then added 10 渭 g / mL LPS for 24 h to detect the changes of the above indexes again. In vivo experiment, 50 adult male C57BL/6 mice were randomly divided into control group, model group and PQ401 group with 10 rats in each group. The model of acute inflammatory reaction was established by intraperitoneal injection of LPS 10 mg/kg. The control group was treated with (DMSO) 10 mg/kg and PQ401 2550100 mg / kg respectively. The levels of serum TNF- 伪, IL-1 尾 and IL-6, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBil) were determined by ELISA 12 h after administration, and the levels of serum alanine aminotransferase (ALT), aspartate transaminase (AST) and total bilirubin (TBil) were measured by automatic biochemical analyzer. The nuclear transcription factor-魏 B p65 (NF- 魏 B p65) was detected by Western blot. Results compared with the group treated with LPS alone, PQ401 decreased the level of TNF- 伪, IL-1 尾 and IL-6 in the supernatant of RAW264.7 cells in a concentration-and time-dependent manner. In the acute peritonitis model induced by LPS, the mice showed mental retardation and decreased activity after 12 h of LPS injection, while the PQ401 group showed the mental state of the mice. The activity was better than that in the model group. PQ401 could effectively reduce the content of TNF- 伪, IL-1 尾 and IL-6 in the serum of mice with acute peritonitis induced by LPS, decrease the level of TNF- 伪, IL-1 尾 and NF- 魏 B p65 in liver tissue. Conclusion PQ401 can inhibit the secretion of inflammatory factors induced by LPS in vivo and in vitro.
【作者单位】：重庆医科大学基础医学院生物化学与分子生物学教研室;第三军医大学大坪医院野战外科研究所一室创伤、烧伤与复合伤国家重点实验室;重庆医科大学附属第二医院肝胆外科;
【基金】：国家自然科学基金面上项目(81372088)~~
【分类号】：R969

【参考文献】