红毛五加多糖对大鼠肝细胞免疫损伤的保护作用及其机制  

Protective effects of polysaccharide AHP-Ⅱ on hepatocyte injury in rats and its mechanism

作　　者：李青青 吴疆 郑碧荣 李儒月 王书贤 朱耀强 LI Qingqing;WU Jiang;ZHENG Birong;LI Ruyue;WANG Shuxian;ZHU Yaoqiang(Weifang Medical University,Weifang 261053, Shangdong, China)

机构地区：[1]潍坊医学院

出　　处：《广西植物》2019年第7期880-886,共7页Guihaia

基　　金：国家自然科学基金(81303198)~~

摘　　要：红毛五加属于五加科植物,而五加科植物大多具有调节免疫力的双向调节作用,红毛五加多糖是从中药红毛五加中提取的单一组分,目前关于红毛五加多糖的抑炎作用还少有报道。为了探究红毛五加多糖AHP-Ⅱ对LPS导致的大鼠肝细胞免疫损伤的保护作用及其作用机制,该研究将实验分为空白组、模型组(LPS,40μg·mL^-1)和AHP-Ⅱ低、中、高剂量组(25,50,100μg·mL^-1)五组,以LPS(40μg·mL^-1)来制备大鼠肝细胞免疫损伤模型,先用ELISA方法检测细胞分泌TNF-α的水平及流式检测ROS的含量,来探究AHP-Ⅱ不同剂量对炎症因子的抑制作用,再用western方法检测P-JNK2的蛋白水平来进一步探究AHP-Ⅱ的抑制作用机制。结果表明:低、中、高剂量组的AHP-Ⅱ均可使肝细胞损伤后的TNF-α含量下降。同时,AHP-Ⅱ中、高剂量组均可使损伤后的肝细胞ROS的分泌量下降,且AHP-Ⅱ高剂量组抑制ROS的作用最强。在加入AHP-Ⅱ后,JNK2蛋白的磷酸化水平呈剂量依赖性降低,AHP-Ⅱ高剂量组抑制作用最强。这说明红毛五加多糖AHP-Ⅱ可以通过降低P-JNK2的蛋白含量来抑制炎症因子TNF-α及ROS的水平,以发挥免疫保护作用。Acanthopanax giraldii belongs to the Araliaceae plant, and most of the Araliaceae plants have a two-way regulation effect in regulating immunity.A.giraldii polysaccharide is a single component extracted from the traditional Chinese medicine A. giraldii .The anti-inflammatory effect of polysaccharides has rarely been reported.This study aimed to explore the protective effect and mechanism of polysaccharide AHP-Ⅱ on LPS-induced hepatocyte injury in rats.This experiment was divided into five groups: blank group, model group (LPS, 40 μg·mL^-1 ), AHP-Ⅱ low, moderate, high-dose group (25, 50,100 μg·mL ^-1 ).LPS (40 μg·mL ^-1 ) was used to prepare the rat hepatocyte immune injury model.The cytotoxic effect of AHP-Ⅱ on hepatocytes was first examined by CCK-8 method to eliminate interference, and the secreted level of TNF-α by the cells and the level of ROS were measured by ELISA and flow cytometry respectively to establish the hepatocyte immune injury model.Then the inhibitory effect of different doses of AHP-Ⅱ on inflammatory factors was explored, and the level of JNK2 protein and its phosphorylation were detected by western method to further explore the inhibitory mechanism of AHP-Ⅱ.The results showed that the experimental dose of AHP-Ⅱ had no damage to cells by CCK-8 method, which indicates that the reduction of inflammatory factors secreted by cells treated by AHP-Ⅱ is not due to its cytotoxic effect.AHP-Ⅱ groups (25, 50,100 μg·mL^-1 ) decreased TNF-α content durig hepatocyte injury.At the same time, the high-dose AHP-Ⅱ group can reduce the secretion of ROS durig hepatocytes injury, and the high-dose AHP-Ⅱ group has the strongest effect in inhibiting ehe conrent of ROS.After the addition of AHP-Ⅱ, the phosphorylation level of JNK2 protein decreased in a dose- dependent manner, and the AHP-Ⅱ high-dose group had the strongest inhibitory effect.This indicate that the polysaccharide AHP-Ⅱ can play an immunoprotective role by reducing the protein content of P- JNK2 and inhibiting the le

关 键 词：红毛五加多糖 脂多糖 肝损伤 JNK 

分 类 号：Q946.[生物学—植物学] R285[医药卫生—中药学]