新型纳米载体Ac-αCD携带的Bcl-xl反义寡核苷酸对肺动脉平滑肌细胞增殖和凋亡的作用  

Effect of a novel nanosystem of Ac-αCD encapsulating Bcl-xl antisense oligonucleotide on proliferation and apoptosis in pulmonary arterial smooth muscle cells

作　　者：刘雪萍[1] 贺斌峰[1] 陈华萍[1] 孙欢[1] 杨俊俊[1] 魏征华[1] 窦寅[2] 王关嵩[1] 

机构地区：[1]第三军医大学新桥医院全军呼吸内科研究所,全军呼吸病研究重点实验室,重庆400037 [2]第三军医大学药学院药剂学教研室,重庆400038

出　　处：《第三军医大学学报》2013年第9期846-849,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(81028001)~~

摘　　要：目的研究纳米载体Ac-αCD携带的Bcl-xl反义寡核苷酸(antisense oligonucleotide,ASON)对大鼠肺动脉平滑肌细胞(rat pulmonary arterial smooth muscle cells,RPASMCs)增殖和凋亡作用。方法设计合成5'端标记Cy3的Bcl-xlASON,由纳米载体Ac-αCD携带。实验分3组:纳米载体携带的Bcl-xl ASON组(ASON-NPs组)、单纯纳米载体组(NPs组)和空白对照组,分别使用纳米载体Ac-αCD携带的Bcl-xl ASON、纳米载体Ac-αCD和培养液处理RPASMCs 48 h,激光共聚焦显微镜观察RPASMCs对纳米载体携带的Bcl-xl ASON的摄取情况;RT-PCR、Western blot检测Bcl-xl的mRNA和蛋白表达;MTT检测处理后细胞的增殖抑制率;流式细胞仪检测细胞凋亡率。结果激光共聚焦显微镜下可见ASON-NPs组细胞质内大量呈颗粒状均匀分布的红色荧光物质,空白对照组和NPs组细胞细胞质内未见红色荧光物质;ASON-NPs组处理的RPASMCs的Bcl-xl mRNA和蛋白表达显著低于空白对照组和NPs组(P<0.05);ASODN-NPs组、NPs组、空白对照组细胞抑制率分别为:(53.61±3.02)%、(6.30±1.90)%、(1.40±0.62)%,凋亡率分别为:(53.04±2.09)%、(10.98±2.03)%、(2.19±0.11)%、ASON-NPs组和NPs组细胞抑制率、凋亡率均显著高于空白对照组(P<0.01),ASON-NPs组均显著高于NPs组(P<0.01)。结论纳米载体Ac-αCD携带的Bcl-xl反义寡核苷酸能被RPASMCs有效摄取,从而抑制其增殖,促进凋亡。Objective To determine the effect of an Ac-αCD nanosystem encapsulating Bcl-xi antisense oligonucleotide （ASON） on the proliferation and apoptosis in pulmonary arterial smooth muscle cells. Methods Bcl-xl ASON that had been hallmarked with the Cy3 in 5＇-end was synthesized, and then encapsu- lated into the nanosystem Ac-αCD. Primarily cultured SD rat pulmonary arterial smooth muscle cells were treated by Ac-αCD-Bcl-xl ASON or Ac-αCD for 48 h, and the cells without nanosystem served as control. Confocal microscopy was employed to observe the taking of the nanosystem by the cells. Expression of Bcl-xl at mRNA and protein levels, grow inhibitory rate and apoptotic rate were detected by RT-PCR and Western blotting, M3F assay and flow eytometry, respectively. Results There were a great deal of brilliantly red-fluo- rescent granules distributed evenly in the cytoplasm in the cells treated by Ac-αCD-Bel-xl ASON. No such red- fluorescent granule was seen in the other 2 kinds of cells. The expression of Bcl-xl at mRNA and protein levels were significantly lower in the cells treated by Ac-αCD-Bel-xl ASON （ P 〈 O. 05 ）. Cell growth inhibitory rate was （53.61 ± 3.02） % in Ac-αCD-Bel-xl ASON, （6.30 ± 1.90） % in Ac-αCD and （ 1.40 ± 0.62） % in control cells. Cell apoptotie rate was （53.04 ± 2.09）% in Ae-αCD-Bcl-xl ASON, （10.98 ± 2.03 ）% in Ac-αCD and （2.19±0.11 ）% in control cells, with the former 2 kinds of cells significantly higher than control （P 〈 0.01 ）, so Ac-αCD-Bel-xl ASON cells than Ac-αCD cells （P 〈 0.01 ）. Conclusion Our Ac-αCD nanosystern encapsulating Bcl-xl ASON is effectively taken by rat pulmonary arterial smooth muscle cells, and then inhibits cell growth and induces cell apoptosis.

关 键 词：纳米载体 BCL-XL 反义寡核苷酸 肺动脉平滑肌细胞 

分 类 号：R322.121[医药卫生—人体解剖和组织胚胎学] R329.28[医药卫生—基础医学]