清化肠饮对脂多糖诱导的人肠上皮细胞NF-κB激活的影响  

Qinghuachang Decoction Inhibited NF-kB Activation in LPS-induced Human Enterocytes

作　　者：陈锦团[1] 柯晓[2] 张歆[3] 方文怡[2] 杨春波[2] 彭军[4] 陈友琴[5] THOMAS J.SPEERRA 

机构地区：[1]福建中医药大学中西医结合学院福州350122 [2]福建中医药大学附属第二人民医院福建省脾胃研究室福州350003 [3]福建中医药大学临床实践教学中心福州350122 [4]福建中医药大学中西医结合研究院福州350122 [5]美国凯斯西储大学医学院彩虹婴幼儿医院克利夫兰俄亥俄州44106

出　　处：《中国中西医结合杂志》2015年第11期1356-1360,共5页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金面上项目(No.81173432);福建省自然科学基金项目(No.2011J01194)

摘　　要：目的运用脂多糖(lipopolysaccharide,LPS)刺激分化的结肠癌Caco-2细胞作为一种人体外肠上皮细胞的炎症模型,探讨清化肠饮的抗炎作用及其分子机制。方法制备清化肠饮,培养人结肠上皮细胞Caco-2,应用E LISA方法测定肿瘤坏死因子-α(TNF-α)和白细胞介素-8(lL-8),Western blot分析法检测抑制性卡巴蛋白(inhibitory Kaba protein,IκB-α)、磷酸化抑制性卡巴蛋白(phosphory lated inhibitory Kaba protein,p-IκB-α)、核转录因子p50(nuclear transcriotion factor p50,p50)、核转录因子RelA(nuclear transcription factor ReIA,RelA)蛋白。结果与未给予LPS刺激比较,LPS刺激能诱导Caco-2细胞TNF-α和IL-8的释放(P<0.05)。清化肠饮处理可降低LPS诱导的TNF-α和IL-8的分泌,并呈剂量依赖性(P<0.05)。0、5、10、50μg/mL不同浓度清化肠饮对Caco-2细胞存活率无明显影响,各浓度间比较,差异无统计学意义(P>0.05)。清化肠饮可抑制LPS刺激后的IκB-α的磷酸化,p-IκB-α的表达情况随着清化肠饮浓度的增加而下降(P<0.05),IκB-α无明显变化(P>0.05)。p50、RelA表达水平随着清化肠饮浓度的增加而下降(P<0.05),均呈剂量依赖性。结论清化肠饮抑制LPS介导的NF-κB活化可能是其治疗IBD的机制之一。Objective To explore anti-inflammation and mechanism of Qinghuachang Decoction（QD）by using LPS stimulated differentiated colon cancer Caco-2 cells（as an inflammation model of human enterocytes）.Methods QD was prepared.Human colonic epithelial Caco-2 cells were cultured.Expressions of TNF-α and IL-8 were determined using ELISA.Expressions of inhibitory Kaba protein（IκB-α）,phosphorylated inhibitory Kaba protein（p-IκB-α）,nuclear transcription factor p50（p50）,and nuclear transcription factor ReIA（ReIA）protein were determined by Western blot.Results Compared with the negative control group（without LPS stimulation）,LPS stimulated the release of IL-8 and TNF-α in Caco-2 cells（P0.05）.QD treatment could reduce the secretion of TNF-α and IL-8 induced by LPS in a dose dependent manner（P0.05）.QD at 0,5,10,and 50 μg/mL had no significant effect on Caco-2 cell survival rates（P0.05）,with no statistical difference among various concentrations（P0.05）.QD could significantly suppress nuclear factor-kappa B（NF-κB） phosphorylation stimulated by LPS.The expression of p-IκB-αwas decreased with increasing concentrations of QD（P0.05）.There was no obvious change in kB-α expressions（P0.05）.Expressions of p50 and RelA decreased with increasing concentrations of QD（P 0.05）.Both of them were in a dose dependent manner.Conclusion QD inhibited LPS mediated NF-κB activation,which might be one of its mechanisms for treating inflammatory bowel disease（IBD）.

关 键 词：清化肠饮 炎症性肠疾病 核转录因子-κB通路 

分 类 号：R285[医药卫生—中药学]