胰岛素对高糖环境中巨噬细胞表型转换的影响

发布时间：2018-06-05 13:12

本文选题：巨噬细胞表型 + 胰岛素　；参考：《上海交通大学学报(医学版)》2017年05期

【摘要】：目的·研究胰岛素对高糖培养的人单核细胞株THP-1细胞及糖尿病创面巨噬细胞表型转换的影响。方法·分别用正常糖(5.6 mmol/L)和高糖(25 mmol/L)培养THP-1细胞,PMA刺激贴壁后,LPS诱导分化为M1型巨噬细胞,胰岛素处理细胞6 h。使用RT-PCR、Western blotting检测M1型标记物诱导型一氧化氮合酶(iNOS)、肿瘤坏死因子α(TNF-α)、白介素-1β(IL-1β)以及M2型标记物精氨酸酶1(Arg1)、IL-10表达的变化。高脂饮食喂养联合小剂量链脲佐菌素(STZ)多次腹腔注射诱导2型糖尿病大鼠模型;血糖稳定5周后,在糖尿病大鼠背部制作2个直径1 cm的全层皮肤缺损创面,应用随机数字表法将创面随机分为胰岛素处理创面和生理盐水处理创面,分别滴加0.2 U胰岛素/20μL生理盐水或20μL生理盐水。于伤后第3、7、25日,观察创面巨噬细胞表型特征;以正常大鼠(n=3)作为正常对照。结果·经高糖培养后,由THP-1细胞诱导分化的巨噬细胞M1型标记物iNOS、TNF-αmRNA表达上调,而M2型标记物Arg1、IL-10 mRNA表达下调;胰岛素作用6 h后,iNOS、TNF-αmRNA表达下调,iNOS、IL-1β蛋白表达也下调,而Arg1、IL-10 mRNA及蛋白表达均上调。此外,高糖培养的巨噬细胞磷酸化NF-κB-p65水平明显升高,而胰岛素干预后磷酸化NF-κB-p65水平显著降低。与正常对照大鼠皮肤创面相比,糖尿病大鼠创面巨噬细胞iNOS表达明显升高;伤后第3、7日,生理盐水处理创面巨噬细胞iNOS高表达;伤后第25日,Arg1表达较低。伤后第7日起,胰岛素处理创面巨噬细胞iNOS表达开始下调;伤后第25日,Arg1的表达显著高于生理盐水处理创面。结论·胰岛素可诱导高糖环境中的巨噬细胞由M1表型向M2表型转换,其机制可能与NF-κB-p65的磷酸化有关。
[Abstract]:Objective to study the effect of insulin on phenotypic transformation of human monocytes THP-1 cells and diabetic wound macrophages cultured with high glucose. Methods THP-1 cells were cultured with normal glucose (5.6 mmol / L) and high glucose (25 mmol / L), respectively. LPS-induced macrophages were induced to differentiate into M1 macrophages and treated with insulin for 6 h. The expression of inducible nitric oxide synthase (iNOSN), tumor necrosis factor- 伪 (TNF- 伪), interleukin-1 尾 (IL-1 尾) and argininase 1Arg1 + IL-10 were detected by RT-PCR- blotting. The rat model of type 2 diabetes was induced by high fat diet and low dose streptozotocin (STZ) intraperitoneal injection. After 5 weeks of blood glucose stabilization, two full-thickness skin defect wounds with diameter of 1 cm were made in the back of diabetic rats. The wounds were randomly divided into insulin treated wounds and saline treated wounds with 0.2 U insulin / 20 渭 L normal saline or 20 渭 L normal saline respectively. The phenotypic characteristics of macrophages were observed on the 3rd and 25th day after injury, and normal rats were used as the normal control. Results after cultured with high glucose, the expression of iNOS- TNF- 伪 mRNA in macrophages induced by THP-1 cells was up-regulated, while the expression of Arg1mIL-10 mRNA was down-regulated, and the expression of iNOS- TNF- 伪 mRNA was also down-regulated after 6 hours of insulin treatment, and the expression of iNOS- 伪 TNF- 伪 mRNA was also down-regulated. The expression of IL-10 mRNA and protein were up-regulated. In addition, the level of phosphorylated NF- 魏 B-p65 in macrophages cultured with high glucose increased significantly, while the level of phosphorylated NF- 魏 B-p65 decreased significantly after insulin intervention. Compared with the normal control rats, the expression of iNOS in the wound macrophages of diabetic rats was significantly higher than that in the normal control rats, and the expression of iNOS in the macrophages of the wounds treated with normal saline on the 3rd and 7th day after injury was higher than that in the rats treated with normal saline, and the expression of Arg1 was lower on the 25th day after injury. From the 7th day after injury, the expression of iNOS in insulin-treated wound macrophages began to decrease, and the expression of Arg1 on the 25th day after injury was significantly higher than that of saline treatment. Conclusion Insulin can induce the transformation of macrophages from M1 phenotype to M2 phenotype in high glucose environment, which may be related to the phosphorylation of NF- 魏 B-p65.
【作者单位】：上海交通大学医学院附属瑞金医院烧伤整形科;
【基金】：国家自然科学基金(81270909,81170761) 上海市市级医院新兴前沿技术联合攻关项目(SHDC12014117)~~
【分类号】：R587.1

【相似文献】