姜黄素对血栓刺激肺微血管内皮细胞血栓模型前炎症因子的影响  

Effects of curcumin on pro-inflammatory factors in pulmonary microvascular endothelial cell thrombus model stimulated by thrombus

作　　者：黄立权[1] 邱添 张蓉蓉[1] 方海军 吴煜[1] 王灵聪[1] Huang Liquan;Qiu Tian;Zhang Rongrong;Fang Haijun;Wu Yu;Wang Lingcong(Department of Intensive Care Unit,First Affiliated Hospital of Zhejiang University of Traditional Chinese Medicine,Hangzhou 310006,Zhejiang,China;Qingbo Community Health Service Center,Shangcheng District,Hangzhou 310006,Zhejiang,China)

机构地区：[1]浙江中医药大学附属第一医院ICU,浙江杭州310006 [2]杭州市上城区清波街道社区卫生服务中心,浙江杭州310006

出　　处：《中国中西医结合急救杂志》2019年第3期269-273,共5页Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care

基　　金：浙江省自然科学基金(LY17H290006, LY12H29005);浙江省卫生高层次创新人才培养工程项目(2014-108).

摘　　要：目的观察姜黄素对血栓刺激肺微血管内皮细胞(LMVEC)中前炎症因子的影响.方法取LMVEC,按随机数字表法分为6组.空白对照组不给予任何处理;LMVEC组用普通培养基培养LMVEC7h;短发夹(shRNA)组用shRNA腺病毒感染细胞72h;不规则趋化因子(CX3CL1)过表达组用CX3CL1过表达腺病毒感染细胞72h;shRNA+姜黄素组用shRNA腺病毒与40μmol/L姜黄素共同处理LMVEC72h;CX3CL1过表达+姜黄素组用CX3CL1过表达腺病毒与40μmol/L姜黄素共同处理LMVEC72h.各组再加入血栓自然沉淀12h.观察每组细胞白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)的含量和CX3CL1、CX3CL1受体(CX3CR1)、IL-6、TNF-α的mRNA表达以及CX3CL1/CX3CR1、CX3CL1/核转录因子-κB(NF-κB)的蛋白表达水平,每组重复3次.结果LMVEC组IL-6、TNF-α含量和mRNA表达以及CX3CR1、NF-κB蛋白表达均较空白对照组明显升高〔IL-6(ng/L):207.90±16.69比85.93±20.32,TNF-α(ng/L):239.60±15.27比101.23±11.92;IL-6mRNA:0.66±0.05比0.11±0.02,TNF-αmRNA:1.06±0.04比0.02±0.01;CX3CR1蛋白:3.94±0.58比1.00±0.31,NF-κB蛋白:1.20±0.07比1.00±0.10;均P<0.05〕;shRNA组、CX3CL1过表达组、shRNA+姜黄素组、CX3CL1过表达+姜黄组IL-6含量均较LMVEC组降低(ng/L:183.60±11.52、159.27±15.02、117.03±7.91、119.97±11.43比207.90±16.69,均P<0.05);TNF-α含量除shRNA组较LMVEC组明显升高外(ng/L:282.00±5.63比239.6±15.27),CX3CL1过表达组、shRNA+姜黄素组、CX3CL1过表达+姜黄素组TNF-α含量均较LMVEC组明显降低(ng/L:216.97±9.20、203.97±19.03、191.97±17.50比239.6±15.27,均P<0.05).CX3CL1过表达组、CX3CL1过表达+姜黄素组CX3CL1的mRNA表达水平均较空白对照组和LMVEC组显著增高(CX3CL1mRNA:55210.3±1209.2、165296.3±8082.4比3.3±0.6、2.0±0.0,均P<0.01).shRNA组IL-6mRNA表达水平较LMVEC组明显升高(IL-6mRNA:0.82±0.17比0.66±0.05),CX3CL1过表达组IL-6mRNA表达水平较LMVEC组明显降低(IL-6mRNA:0.29±0.03比0.66±0.05),加入姜黄素预处理后的变化更�Objective To explore the effects of curcumin on pro-inflammatory factors in the lung microvascular endothelial cells (LMVEC) model stimulated by thrombus. Methods The LMVECs were divided into six groups according to the random number table method. No treatment was given to the blank control group;the model group was cultured for 7 hours in normal medium;the curcumin group was treated with 40 μmol/L curcumin for 72 hours;the shRNA group was infected with shRNA adenovirus for 72 hours;the irregular chemokines (CX3CL1) overexpression group was infected with CX3CL1 overexpressing adenovirus for 72 hours;the shRNA+curcumin group infected with shRNA adenovirus and treated with 40 μmol/L curcumin together for 72 hours;CX3CL1 overexpression +curcumin group infected with CX3CL1 overexpressing adenovirus and treated with 40 μmol/L curcumin together for 72 hours. After each group was given the corresponding pretreatment, the thrombus natural precipitation was added each group for 12 hours. The contents of interleukin-6 (IL-6) and tumor necrosis factor-α(TNF-α), the mRNA expression levels of CX3CL1, CX3CL1 receptor (CX3CR1), IL-6, TNF-α and the protein expression levels of CX3CL1/CX3CR1, CX3CR1/NF-κB in various groups were observed, repeat 3 times in each group. Results The contents and mRNA expression of IL-6, TNF-αand protein expression of CX3CR1, NF-κB in the LMVEC group were significantly higher than those in blank control group [IL-6 (ng/L): 207.90±16.69 vs. 85.93±20.32, TNF-α(ng/L): 239.60±15.27 vs. 101.23±11.92;IL-6 mRNA: 0.66±0.05 vs. 0.11±0.02, TNF-α mRNA: 1.06±0.04 vs. 0.02±0.01;CX3CR1 protein:3.94±0.58 vs. 1.00±0.31, NF-κB protein: 1.20±0.07 vs. 1.00±0.10;all P < 0.05];the contents of IL-6 in shRNA group, CX3CL1 overexpression group, shRNA + curcumin group, CX3CL1 overexpression + curcumin group were all obviously lower than those in LMVEC group (ng/L: 183.60±11.52, 159.27±15.02, 117.03±7.91, 119.97±11.43 vs. 207.90±16.69, all P < 0.01);the content of TNF-α was markedly increased in s

关 键 词：姜黄素 肺微血管内皮细胞 白细胞介素-6 肿瘤坏死因子-α 核转录因子-ΚB 不规则趋化因子 

分 类 号：R563[医药卫生—呼吸系统]