槐定碱对LPS诱导的RAW264.7巨噬细胞CD14、P38、iNOS表达的影响  

Effects of sophoridine on lipopolysaccharide-induced expressions of CD14,iNOS and P38 in RAW264.7 cells

作　　者：张巍[1,2] 李业成[3] 张高明[2] 周娅[1] 

机构地区：[1]宁夏医科大学基础医学院病原生物学与医学免疫学系,银川750004 [2]沭阳县人民医院检验科,宿迁223600 [3]沭阳县人民医院骨科,宿迁223600

出　　处：《免疫学杂志》2015年第2期100-104,共5页Immunological Journal

基　　金：国家自然科学基金(30660227);宁夏自然科学基金(HZ0783)

摘　　要：目的探讨槐定碱不同给药方式对内毒素诱导的小鼠RAW264.7巨噬细胞表达CD14、p38 MAPK及i NOS的影响及意义。方法采用LPS诱导的RAW264.7巨噬细胞建立细胞炎症反应模型;实验细胞分为4组(n=6):空白对照组、LPS组、槐定碱预处理组、槐定碱预混合组;分别利用反转录聚合酶链反应(RT-PCR)和Western blot技术检测RAW264.7巨噬细胞CD14、p38、i NOS m RNA表达量及蛋白表达量。结果槐定碱2种给药方式(预处理及预混合)均可显著下调LPS诱导的RAW264.7巨噬细胞p38、i NOS m RNA及CD14、p-p38、i NOS蛋白表达;槐定碱与LPS预混合可下调LPS诱导的RAW264.7巨噬细胞CD14 m RNA表达,而槐定碱预处理对CD14 m RNA表达无明显影响。结论槐定碱可能通过调控CD14、p38、i NOS表达与活化发挥抗内毒素效应。To investigate the roles of sophoridine（SRI） in inflammation induced by lipopolysaccharide（LPS） in two different modes of administration,LPS-activated macrophage cells（RAW264.7） was employed as an inflammation model. RAW264.7 cells were divided into 4 groups（n=6）： control group,LPS group,SRI pretreating group,SRI premixing group. The m RNA and protein expressions of CD14,p38,i NOS were measured by using reverse transcription polymerase chain reaction（RT-PCR） and Western blotting,respectively. Data showed that compared with LPS group,the expressions of CD14,p-p38 and i NOS protein and p38,i NOS m RNA were significantly decreased in SRI pretreating group and SRI premixing group（P〈0.01）,which indicated that SRI can inhibit the expression of CD14,p-p38 and i NOS protein and p38,i NOS m RNA induced by LPS in RAW264.7 cells.Furthermore,compared with SRI pretreating group,the expression of CD14 m RNA was significantly decreased in SRI premixing group（P〈 0.01）,which indicated that SRI premixing group can abolish expression of CD14 m RNA.Taken together,SRI can play anti-endotoxin role in inflammation by decreasing expressions of CD14,p-p38,i NOS protein and p38,i NOS m RNA in TLR4 pathway.

关 键 词：槐定碱 脂多糖 RAW264.7巨噬细胞 

分 类 号：R285[医药卫生—中药学]