RyR反义寡核苷酸对气道平滑肌细胞增殖及细胞内钙离子浓度的影响

目的:观察RyR(Ryanodme受体)反义寡核苷酸(ASODN)对大鼠ASMCs(airway smooth muscle cells,气道平滑肌细胞)增殖的抑制作用及对细胞内钙离子浓度的影响.方法:采用胶原酶消化法培养大鼠ASMCs,利用LipofectamineTM2000将正义、反义RyR寡核苷酸导入大鼠ASMCs,MTS/PES法检测不同寡核苷酸对大鼠ASMCs增殖的抑制作用,RT-PCR检测大鼠ASMCs中RyR的mRNA表达,流式细胞仪测定不同寡核苷酸对细胞内钙离子浓度的影响.结果:RyR反义寡核苷酸可抑制大鼠ASMCs的增殖,降低其RyR受体mRNA的表达,并能降低兴奋后的细胞内钙离子浓度的升高.结论:RyR反义寡核苷酸可能通过降低兴奋后的细胞内钙离子浓度来抑制大鼠ASMCs的增殖.

Effect of antisense oligonucleotides of ryanodine receptor on proliferation and [Ca2+]i of airway smooth muscle cells

Aim: To investigate the effect of antisense oligonucleotides(ASON) of ryanodine receptor on proliferation and Ca~(2 )]i concentration of airway smooth muscle cells(ASMCs). Methods: ASMCs were cultivated with collagen enzyme digestion method. Different concentrations of ASON were added to the cultures with Lipofectamine (~(TM) 2000) to observe the ASMCs proliferation using MTS/PES method. The changes of ASMCsCa~(2 )]i were also observed by flow cytometry. The expression of mRNA of subtyes of RyR was asssyed by RT-PCR. Results: RyR ASON restrained the proliferation of ASMCs, decreased the expression of RyR and reduced the concentration of Ca~(2 )]i. Conclusion: RyR ASON could inhibit the proliferation of ASMCs by influencing the concentration of Ca~(2 )]_i after excited.