基于β-咔啉的环金属化Ru(Ⅱ)配合物诱导HeLa细胞凋亡及抗氧化活性研究  

The apoptosis-inducing properties and antioxidant activity of cyclometalated Ru(Ⅱ)-β-carboline complexes

作　　者：陈锦灿 蔡先洪 彭发 陈李祺 孙梓荣 罗辉 陈兰美 CHEN Jin-can;CAI Xian-hong;PENG Fa;CHEN Li-qi;SUN Zi-rong;LUO Hui;CHEN Lan-mei(Southern Marine Science and Engineering Guangdong Laboratory,Zhanjiang 524025,China;The Marine Biomedical Research Institute,Guangdong Medical University,Zhanjiang 524023,China;The Marine Biomedical Research Institute of Guangdong Zhanjiang,Zhanjiang 524023,China;School of Pharmacy,Guangdong Medical University,Zhanjiang 524023,China)

机构地区：[1]南方海洋科学与工程广东省实验室,广东湛江524025 [2]广东医科大学海洋医药研究院,广东湛江524023 [3]广东湛江海洋医药研究院,广东湛江524023 [4]广东医科大学药学院,广东湛江524023

出　　处：《中国海洋药物》2022年第6期31-40,共10页Chinese Journal of Marine Drugs

基　　金：国家自然科学基金项目(21701034);广东省自然科学基金项目(2020A1515010444);南方海洋科学与工程广东实验室(湛江)项目(ZJW-2019-007);广东医科大学学科建设项目(4SG21004G);2021年广东省大学生创新计划项目(GDMU2021063)资助。

摘　　要：目的探究新型环金属化Ru(Ⅱ)-β-咔啉配合物[Ru(dmb)_(2)(NO_(2)-Ph-βC)](PF6)(NO_(2)-Ph-βC=NO_(2)-phenyl-9H-pyrido[3,4-b]indole,RuβC-5)和[Ru(bpy)2(NO_(2)-Ph-βC)](PF6)(RuβC-6)诱导He La细胞凋亡的机理。方法通过元素分析、ES-MS、^(1)H NMR对配合物进行表征;采用电感耦合等离子体质谱(ICP-MS)测定配合物的油水分配系数log P_(o/w);细胞毒性实验检测RuβC-5和RuβC-6对A549、He La、NCI-H460、HepG-2和MCF-7细胞的毒性作用;Hoechst 33342、吖啶橙/溴化乙锭(AO/EB)、Annexin V-FITC/PI染色法检测配合物诱导He La细胞凋亡;流式细胞术分析配合物对He La细胞凋亡、细胞周期阻滞、线粒体膜电位(MMP)、活性氧(ROS)的影响,蛋白免疫印迹实验(Western Blot)检测Caspase信号通路及线粒体凋亡通路相关蛋白的影响。结果配合物RuβC-5、RuβC-6对He La细胞表现出相对较高的细胞毒性和较好的选择性,能诱导He La细胞的G0/G1期阻滞和凋亡,降低MMP,抑制细胞内ROS的产生。结论新型Ru(Ⅱ)-β-咔啉配合物可诱导He La细胞凋亡;此外,RuβC-5具有抗氧化活性,可以通过清除细胞内ROS的含量诱导He La细胞凋亡。Objective To investigate the mechanism of HeLa cell apoptosis-inducing by two cyclometalated Ru(Ⅱ)-β-carboline complexes[Ru(dmb)_(2)(NO_(2)-Ph-βC)](PF6)(NO_(2)-Ph-βC=NO_(2)-phenyl-9H-pyrido[3,4-b]indole,RuβC-5)and[Ru(bpy)2(NO_(2)-Ph-βC)](PF6)(RuβC-6).Methods The complexes were characterized by elemental analysis,ES-MS,^(1)H NMR and UV-Vis spectra.The oil-water partition coefficient of complexes were measured by using inductively coupled plasma mass spectrometry(ICP-MS).The cytotoxicity of RuβC-5 and RuβC-6 were assessed against A549,HeLa,NCI-H460,HepG-2 and MCF-7 cells.Their effects on apoptosis,cell cycle,mitochondrial membrane potential(MMP)and reactive oxygen species generation(ROS)of A549 were determined using?ow cytometry.Western Blot assay was used to detect the impact of Caspase signaling pathway and mitochondrial apoptosis pathway related proteins.Results The complexes RuβC-5 and RuβC-6 exhibited much higher cytotoxicity and good selectivity to normal cells,which could induce G0/G1phase arrest and apoptosis,decrease MMP and inhibit the production of intracellular ROS.Conclusion The new Ru(Ⅱ)-β-carboline complexes could induce HeLa cells apoptosis.Furthermore,as an antioxidant,RuβC-5tended to induce apoptosis in HeLa cells by scavenging intracellular ROS.

关 键 词：β-咔啉生物碱 环金属化Ru(Ⅱ)配合物 细胞凋亡 细胞毒性 抗氧化活性 

分 类 号：R914[医药卫生—药物化学]