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Aim:To investigate the effect of 5-hydroxytryptamine transporter(5-HTT)inhibi-tor fluoxetine and antisense oligodeoxynucleotide(ODN)to extracelluar signal-regulated kinases(ERKs)on pulmonary arterial smooth muscle cells(PASMCs)proliferation induced by 5-HT.Methods:Liposomal transfection was used tointroduce ODNs to ERK1/2 into cultured rat PASMCs and the transfection effi-ciency was measured by observing the uptake of the fluorecein isothiocynate(FITC)-labeled antisense ODN in PASMCs.The effects of 5-HTT selective inhibi-tor fluoxetine and ODNs on the proliferation of PASMCs were evaluated by cellnumber counting and cell cycle analysis,and measured by microculture tetrazo-lium(MTT)assay and flow cytometry(FCM),respectively.Results:Liposomesmediated the transfection of ODNs into PASMCs with high efficiency.MTTassay showed fluoxetine(10μmol/L,1 μmol/L,and 100 nmol/L)concentrationdependently inhibited the proliferation of PASMCs induced by 5-HT(1 μmol/L)invitro.The proliferation rate of PASMCs by 5-HT was significantly inhibited bypretreatment with ERK1/2 antisense ODN(0.2μmol/L)from 251%±18% to 86%±5%(P<0.01).Flow cytometric analysis of cell cycle distribution showed that theincrease of 5-HT induced S phase fraction(SPF)and proliferation index(PI)weresignificantly inhibited by fluoxetine(1μmol/L)or antisense ODN with SPF from36%±4% to 26%±3% and 24%±4%,and PI from 34%±2% to 29%±2% and 24%±2%,respectively.Conclusion:5-HTT mediates the mitogenic effect of 5-HT onPASMCs and the proliferation of PASMCs induced by 5-HT is dependent onERKs signal pathway.
Aim: To investigate the effect of 5-hydroxytryptamine transporter (5-HTT) inhibi-tor fluoxetine and antisense oligodeoxynucleotide (ODN) to extracellular signal-regulated kinases (ERKs) on pulmonary arterial smooth muscle cells (PASMCs) . Methods: Liposomal transfection was used to introduce ODNs to ERK1 / 2 into cultured rat PASMCs and the transfection effi- ciency was measured by observing the uptake of the fluorecein isothiocynate (FITC) -labeled antisense ODN in PASMCs.The effects of 5-HTT selective inhibi-tor fluoxetine and ODNs on the proliferation of PASMCs were evaluated by cell number counting and cell cycle analysis, and by by microculture tetrazo-lium (MTT) assay and flow cytometry (FCM), respectively. Results: Liposomesized the transfection of ODNs into PASMCs with high efficiency. MTT assay showed fluoxetine (10 μmol / L, 1 μmol / L, and 100 nmol / L) concentration dependently inhibited the proliferation of PASMCs induced by 5-HT f PASMCs by 5-HT was significantly inhibited from pretreatment with ERK1 / 2 antisense ODN (0.2 μmol / L) from 251% ± 18% to 86% ± 5% (P <0.01) of 5-HT induced S phase fraction (SPF) and proliferation index (PI) weresignificantly inhibited by fluoxetine (1 μmol / L) or antisense ODN with SPF from 36% ± 4% to 26% ± 3% and 24% ± 4% PI from 34% ± 2% to 29% ± 2% and 24% ± 2%, respectively.Conclusion: 5-HTT mediates the mitogenic effect of 5-HT onPASMCs and the proliferation of PASMCs induced by 5-HT is dependent onERKs signal pathway.