钴60γ射线辐射对烟肼酰胺的结构及抗炎活性的影响  

Effects of ^(60)Co-γradiation on the structure and anti-inflammatory activity of nialamide

作　　者：严鹏 侯静 李平 陈蕊蕊 李妍 YAN Peng;HOU Jing;LI Ping;CHEN Ruirui;LI Yan(Department of Cardiovascular Medicine,The Second Affiliated Hospital of Air Force Medical University,Xi'an 710000 China;Department of Clinical Medicine,Xi'an Public Health Center,Xi'an 710000 China;Department of Pharmacy,Xi'an Hospital of Traditional Chinese Medicine,Xi'an 710000 China)

机构地区：[1]空军军医大学第二附属医院心血管内科,陕西西安710000 [2]西安市公共卫生中心临床药学部,陕西西安710000 [3]西安市中医医院药剂科,陕西西安710000

出　　处：《中国辐射卫生》2024年第5期491-498,548,共9页Chinese Journal of Radiological Health

基　　金：陕西省自然科学基础研究计划面上项目(306232575018);国家自然科学基金(NO.81800346)。

摘　　要：目的研究钴^(60)γ射线(^(60)Co-γ)辐射对烟肼酰胺结构的影响,对比辐照产物的抗炎活性,并探究其作用机制。方法烟肼酰胺在50 kGy剂量下进行^(60)Co-γ辐照后,得到5种已知化合物(2-6),并分别作用于RAW 264.7细胞(小鼠单核巨噬细胞白血病细胞)。采用CCK-8检测RAW 264.7细胞活力;应用酶联免疫吸附法、Griess法测定细胞白介素-6(IL-6)、白介素-10(IL-10)、肿瘤坏死因子-α(TNF-α)、前列腺素E_(2)(PGE_(2))分泌及一氧化氮(NO)含量的变化;采用DCFH-DA荧光探针检测活性氧(ROS)生成;采用Western blot检测细胞诱导型一氧化氮合酶(iNOS)、环氧化酶(COX-2)、核转录因子(NF-κB)、IκB蛋白表达水平的情况。结果烟肼酰胺辐照后得到的产物对RAW264.7细胞活力均无明显影响(P>0.05),但表现出很强的抗炎作用(P<0.01)。与辐照前烟肼酰胺相比,化合物(2、3、4、6)显著降低了LPS诱导的RAW 264.7细胞中NO含量(P<0.01),其中化合物4作用最为显著;同时化合物4显著降低了LPS诱导的RAW 264.7细胞中IL-6、TNF-α、PGE_(2)、ROS的含量(P<0.05),iNOS、COX-2、NF-κB、IκB蛋白表达量降低(P<0.05)。结论烟肼酰胺经^(60)Co-γ辐照后,其化学结构改变,其产物化合物4具有较强的抗炎活性,其作用可能与抑制NF-κB信号通路激活,进而减少炎症因子释放有关。辐射技术为天然产物分子结构和生理特性的改变可提供新的见解。Objective To study the effect of cobalt-60 gamma-ray(^(60)Co-γ)radiation on the structure of Nialamide,com-pare the anti-inflammatory activity of irradiation products,and explore the mechanism of action.Methods After ^(60)Co-γir-radiation of nialamide at a dose of 50 kGy,five known compounds were obtained(2-6).The viability of RAW 264.7(mouse mononuclear macrophage leukemia)cells treated with these compounds was determined by CCK-8 assay.The secretion of interleukin-6(IL-6),interleukin-10(IL-10),tumor necrosis factor-α(TNF-α),and prostaglandin E_(2)(PGE_(2))and the content of nitric oxide(NO)were measured using enzyme-linked immunosorbent assay and Griess method.The production of reactive oxygen species(ROS)was detected using DCFH-DA fluorescent probe.The expression levels of cell-induced nitric oxide synthase(iNOS),cyclooxygenase(COX-2),nuclear transcription factor-κB(NF-κB),and IκB were detected using Western blot.Results The products of nialamide after irradiation did not significantly affect RAW264.7 cell viability(P>0.05)but showed a strong anti-inflammatory effect(P<0.01).Compared with nialamide,compounds 2,3,4,6 significantly reduced NO content in LPS-induced RAW 264.7 cells(P<0.01),and compound 4 had the most significant effect.Moreover,com-pound 4 significantly reduced the content of IL-6,TNF-α,PGE_(2),and ROS(P<0.05)as well as the expression of iNOS,COX-2,NF-κB,and IκB(P<0.05)in LPS-induced RAW 264.7 cells.Conclusion The chemical structure of nialamide is changed after irradiation with ^(60)Co-γ,and its product compound 4 shows strong anti-inflammatory activity,which may be re-lated to inhibiting the activation of NF-κB signaling pathway and reducing the release of inflammatory factors.Radiation technology can provide new insights into the changes of molecular structures and physiological properties of natural products.

关 键 词：电离辐射 烟肼酰胺 ^(60)Co-γ射线 抗炎 RAW 264.7细胞 

分 类 号：X591[环境科学与工程—环境工程]