去氢骆驼蓬碱及其衍生物DH-004暴露对小鼠神经系统毒性作用研究  

Study on the neurotoxic effects of harmine and its derivatives DH-004 in mice

作　　者：茹则妮萨·阿卜杜拉 高惠静[2] 巩月红[2] 文丽梅 高旖 王建华[2] 陈蓓[2] Ruzenisa·abudula;GAO Hui-jing;GONG Yue-hong;WEN Li-mei;GAO Yi;WANG Jian-hua;CHEN Bei(College of Pharmaceutical Sciences,Xinjiang Medical University,Urumqi 830000,China;Department of Pharmacy,First Affiliated Hospital of Xinjiang Medical University,Urumqi 830011,China)

机构地区：[1]新疆医科大学药学院,乌鲁木齐830000 [2]新疆医科大学第一附属医院药学部,乌鲁木齐830011

出　　处：《中国新药杂志》2021年第8期740-746,共7页Chinese Journal of New Drugs

基　　金：国家自然科学基金项目(81860666);省部共建中亚高发病成因与防治国家重点实验室开放课题资助项目(SKL-HIDCA-2019-24);新疆维吾尔自治区自然科学基金(2018D01C191);新疆重大疾病防治与转化协同创新中心资助项目(80910721);兵团重点领域科技攻关计划项目(2020AB028)。

摘　　要：目的:研究去氢骆驼蓬碱(harmine,HM)及其衍生物1-(4-甲氧基)苯基-9-丁基-β-咔啉(DH004)对脑神经毒性的作用。方法:免疫吸附测定法(ELISA)测定小鼠脑组织中IL-1β,TNF-α,IL-12和IL-6含量;尼氏染色法检测小鼠大脑皮质神经细胞形态改变;实时荧光定量逆转录聚合酶链反应法(qRT-PCR)检测脑组织中离子钙接头蛋白(ionized calcium binding adaptor molecule-1,Iba-1)基因的表达水平;蛋白免疫印迹法(Western-Blot)检测脑组织中Iba-1蛋白的表达水平。结果:ELISA实验结果显示,除了IL-6因子在各剂量组的含量变化不明显,其余3个因子HM处理组小鼠脑组织中的表达量最高,给予相同剂量的DH-004后,与HM处理组相比,小脑组织中炎症因子表达出现显著下降(P<0.01)。尼氏染色、qRT-PCR、WesternBlot实验结果显示,与空白组相比,HM给药组对小鼠皮质神经细胞损伤严重,且有剂量依赖性;HM组Iba1mRNA和Iba-1蛋白表达水平较相应的空白组明显上调(P<0.01),给予相同剂量的DH-004后,与HM处理组相比,小脑组织中Iba-1mRNA和Iba-1蛋白表达水平显著下调(P<0.01)。结论:HM可能诱导小鼠小胶质细胞的活化,激活的小胶质细胞促使炎症因子释放增加,导致神经组织损伤;而DH-004可能抑制小胶质细胞活化,从而减少炎症因子释放,减轻神经毒性。Objective:To study the neurotoxicity of DH-004,a derivative of harmine(HM).Methods:IL-1β,TNF-α,IL-12 and IL-6 in the brain of mice were determined by enzyme-linked immunosorbent assay(ELISA).Morphological changes of neurons in cerebral cortex of mice were detected by Nissl staining.Real-time fluorescence quantitative reverse transcription polymerase chain reaction(qRT-PCR)was used to detect ionized calcium binding adaptor molecule-1(Iba-1)mRNA expression in brain tissue.Iba-1 protein expression in brain tissue was detected by Western-Blot.Results:ELISA results showed that except for the insignificant changes of IL-6 in each dose group,the expression levels of the other three inflammatory factors with HM-treated mice in the brain tissues were the highest.After the same dose of DH-004 was given,the expression levels of inflammatory factors in the cerebellum tissues were significantly decreased compared with HM-treated mice(P<0.01).The results of Nisier staining,qRT-PCR and Western-Blot showed that compared with the blank control group,HM group had serious and dose-dependent damage to the cortical nerve cells of mice.The expression levels of Iba-1 mRNA and Iba-1 protein in HM group were up-regulated compared with the corresponding blank control group(P<0.01),after the same dose of DH-004 was given,the expression levels of Iba-1 mRNA and Iba-1 protein in cerebellum tissues were significantly down-regulated compared with HM group(P<0.01).Conclusion:HM can induce the activation of microglia cells in mice,then the activated microglia cells promote the release of inflammatory factors,leading to nerve tissue damage.While DH-004 can inhibit the activation of microglia cells,thus reducing the release of inflammatory factors and reducing neurotoxicity.

关 键 词：DH-004 神经毒性 小胶质细胞 炎症因子 尼氏染色 

分 类 号：R965.3[医药卫生—药理学]