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目的 研究反义磷脂酰肌醇 3 激酶 (PI 3 激酶 )寡脱氧核苷酸 (oligodeoxynucleotides,ODN)对核因子 κB(NF κB)活化的影响。方法 Lipofectin介导反义PI 3 激酶ODN转染HepG2细胞 ,以逆转录PCR法检测PI 3 激酶mRNA表达水平 ,用SandwichELISA法检测NF κB的活化。结果 (1)反义PI 3 激酶ODN抑制PI 3 激酶mRNA表达。 (2 )反义PI 3 激酶ODN呈剂量 (1～ 8μg)和时间 (5～ 2 4h)依赖性地抑制NF κB活化 ,2 μg反义PI 3 激酶ODN与细胞共孵育 8h时抑制效果最好。结论 反义PI 3 激酶ODN通过阻断PI 3 激酶表达抑制NF κB活化 ,预示反义PI 3 激酶ODN可潜在抑制IL 1的炎症活性
Objective To investigate the effects of antisense phosphatidylinositol 3 - kinase (PI 3 - kinase) oligodeoxynucleotides (ODN) on the activation of nuclear factor κB (NF κB). Methods Lipofectin-mediated antisense PI 3 kinase ODN was transfected into HepG2 cells. The expression of PI 3 kinase mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). The activation of NF-κB was detected by Sandwich ELISA. Results (1) Antisense PI 3 kinase ODN inhibited PI 3 kinase mRNA expression. (2) ODN of antisense PI 3 kinase inhibited NF-κB activation in dose (1 ~ 8μg) and time (5 ~ 24h), and ODN of 2 μg antisense PI 3 kinase co-incubated for 8h showed the best inhibitory effect . Conclusion Antisense PI 3 kinase ODN inhibits the activation of NF-κB by blocking the expression of PI 3 kinase, indicating that antisense PI 3 kinase ODN can potentially inhibit the inflammatory activity of IL 1