细柱五加叶中凹脉鹅掌柴酸通过抑制NF-κB-iNOS-NO信号保护胰岛β细胞研究  

Protective Effect of Impressic Acid from the Leaves of Acanthopanax gracilistylus on IsletβCells via Suppression of NF-κB-iNOS-NO Activation

作　　者：鲁曼霞[1] 杨阳 卢茂芳[1] 罗姣 刘向前[1] LU Man-xia;YANG Yang;LU Mao-fang;LUO Jiao;LIU Xiang-qian(School of Pharmacy,Hunan University of Chinese Medicine,Changsha 410208,China;School of Life Sciences,Datong University,Datong 037009,China)

机构地区：[1]湖南中医药大学药学院,长沙410208 [2]大同大学生命科学学院,山西大同037009

出　　处：《中国药学杂志》2022年第12期1008-1014,共7页Chinese Pharmaceutical Journal

基　　金：湖南省教育厅科学研究项目资助(18C0399);湖南省自然科学基金项目资助(2019JJ50448,2019JJ40223);湖南中医药大学生物工程重点学科资助项目资助(校行科字[2018]3号)。

摘　　要：目的基于通过抑制NF-κB-iNOS-NO信号通路,研究细柱五加叶(Acanthopanax gracilistylus W.W.Smith)中凹脉鹅掌柴酸(impressic acid)的胰岛β细胞保护作用。方法运用炎症细胞因子IL-1β和IFN-γ诱导RIN-m5F细胞建立2型糖尿病细胞模型;采用MTT法对细胞毒性进行评价;采用ELISA法测胰岛素分泌评价胰岛细胞功能;采用Griess法测定导致胰岛β细胞的凋亡的关键物质一氧化氮(NO)水平;采用Caspase-3活性检测法检测Caspase-3水平;检测细胞内活性氧物质(reactive oxygen species,ROS)水平;并采用Western blot法检测相关蛋白(iNOS、NF-κB)的表达。结果细柱五加叶中提取的凹脉鹅掌柴酸(5~20μmol·L^(-1))无细胞毒性,并且能够通过抑制NF-κB-iNOS-NO信号通路以浓度依赖的方式恢复细胞因子IL-1β和IFN-γ诱导RIN-m5F细胞凋亡、下调NF-κB表达,下调iNOS表达,降低NO水平,抑制细胞凋亡剪切酶Caspase-3活性,下调活性氧物质ROS水平保护胰岛素β细胞。结论凹脉鹅掌柴酸(5~20μmol·L^(-1))能够通过抑制NF-κB-iNOS-NO信号通路对胰岛β细胞起到保护作用,从而抗2型糖尿病进展。OBJECTIVE To study the protective effect of the compound impressic acid in Acanthopanax gracilistylus W.W.Smith,based on the inhibition of the NF-κB-iNOS-NO signaling pathway.METHODS Inflammatory cytokines IL-1βand IFN-γwere used to induce RIN-m5 F cells as a type 2 diabetic cell model.MTT method was used for cytotoxicity evaluation.ELISA method was used to measure insulin secretion to evaluate islet cell function.Griess method was used to determine the apoptosis of pancreaticβcells The level of nitric oxide(NO),the key substance of death.Caspase-3 activity detection method is used to detect Caspase-3 level;intracellular reactive oxygen species ROS level.Western-blotting method is used to detect related proteins(iNOS,NF-κB)expression.RESULTS Impressic acid(5~20μmol·L^(-1))extracted from A.sibiricus leaves is not cytotoxic and can restore the cytokines IL-1βand IFN-γto induce RIN-m5 F in a concentration-dependent manner by inhibiting the NF-κB-iNOS-NO signaling pathway.Apoptosis,down-regulation of NF-κB expression,down-regulation of iNOS expression,reduction of NO levels,inhibition of the activity of the apoptotic splicing enzyme Caspase-3,and down-regulation of reactive oxygen species ROS levels protect insulinβcells.CONCLUSION Therefore,it is the first report that impressic acid(5~20μmol·L^(-1))inhibits NF-κB-iNOS-NO signal anti-type 2 diabetes and isletβcell protection.

关 键 词：凹脉鹅掌柴酸 RIN-m5F细胞 一氧化氮 caspase-3 活性氧 NF-κB-iNOS-NO 

分 类 号：R965[医药卫生—药理学]