Basic Studies of Novel Gene Therapy by Regulation of Transcription of the Inflammation-Related Gene.

研究novel anti-inflammatory therapies， we studied a novel carrier for antisense DNA，targeted to pro-inflammatory cytokines. We developed pury -l - lessine / l -serine-polyethylene glycol（PLSP） as a carrier for antisense DNA. Our previous study (antisense Nucl Acid Drug Dev 6:55-61，1996) showed that antisense phosphorothioate oligonucleotide targeted to the initiation codon ofhuman interliukin-1β(Il-1β) suppressed production of Il-1β of U937 cells. Then，我们used the antisense DNA of IL-1β to examine the effects of several kinds of PLSP. 10kda and 15kdamolecules of the PSLP were the most potent among them. Howevera considerable toxicity and instability observed when the PSLP was used. We further examinedthe effects of antisense phosphorothioate oligonucleotide targeted to some sequences of human tumornecrosis factor (TNF) α gene. In this case不positive effect was observed. We then studied the anti-inflammatory effects of Tripterygiumwilfordii Hook F extract (GTW). GTW inhibited prostaglandin E - D22 - D2 production in the humansynovial cells due to suppression of cyclooxgenase-2 protein and mRNAwhich is similar to those of glucocorticoid. However，the mechanism of action of the GTW was different from that of glucocorticoid. It inhibited nuclearfactor- D2K D2B activity without acting on glucocorticoid receptor. We also examined effects ofchanges in signal transduction by introduction of Ki-ras gene into human synovial cells frompatients with rheumatoid arthritis. Stimulation by cytokines including TNF-α reduced proliferationrate of ras-introduced synovial cells，whereas他们增强的方案on nontreated synovial cells. The mechanism remains to bestudied。

To investigate novel anti-inflammatory therapies, we studied a novel carrier for antisense DNA, targeted to pro-inflammatory cytokines. We developed poly-L-lysine/L-serine-polyethylene glycol (PLSP) as a carrier for antisense DNA. Our previous study (Antisense Nucl Acid Drug Dev 6:55-61, 1996) showed that antisense phosphorothioate oligonucleotide targeted to the initiation codon of human interliukin-1β(Il-1β) suppressed production of IL-1β of U937 cells. Then, we used the antisense DNA of IL-1β to examine the effects of several kinds of PLSP. 10kDa and 15kDa molecules of the PSLP were the most potent among them. However, a considerable toxicity and instability were observed when the PSLP was used. We further examined the effects of antisense phosphorothioate oligonucleotide targeted to some sequences of human tumor necrosis factor (TNF)-α gene. In this case, no positive effect was observed. We then studied the anti-inflammatory effects of Tripterygium wilfordii Hook F extract (GTW). GTW inhibited prostaglandin EィイD22ィエD2 production in the human synovial cells due to suppression of cyclooxgenase-2 protein and mRNA, which is similar to those of glucocorticoid. However, the mechanism of action of the GTW was different from that of glucocorticoid. It inhibited nuclear factor-ィイD2KィエD2B activity without acting on glucocorticoid receptor. We also examined effects of changes in signal transduction by introduction of Ki-ras gene into human synovial cells from patients with rheumatoid arthritis. Stimulation by cytokines including TNF-α reduced proliferation rate of ras-introduced synovial cells, whereas they enhanced proliferation on nontreated synovial cells. The mechanism remains to be studied.