人主动脉硫酸乙酰肝素蛋白聚糖对培养的人血管壁细胞生长的作用

Effect of Human Aortic Heparan Sulfate Proteoglycan on the Proliferation of Cultured Cells of Human Vascular Wall

通过培养的人主动脉平滑肌细胞（ｈＡＳＭＣ）及脐静脉内皮细胞（ｈＵＶＥＣ），应用３Ｈ－ＴｄＲ参入、Ｎｏｒｔｈｅｒｎｂｌｏｔ分析、逆转录多聚酶链反应（ＲＴ－ＰＣＲ）、放射免疫分析（ＲＩＡ）、和紫外比色法等技术观察了人主动脉中硫酸乙酰肝素蛋白聚糖（ＨＳＰＧ）对ｈＡＳＭＣ和ｈＵＶＥＣＤＮＡ合成的作用及对血小板源生长因子（ＰＤＧＦ）、ＰＤＧＦ受体、转化生长因子β（ＴＧＦ－β）、内皮素－１（ＥＴ－１）或碱性成纤维细胞生长因子（ｂＦＧＦ）基因表达和肾素－血管紧张系统（ＲＡＳ）的影响，结果显示，ＨＳＰＧ明显抑制培养的ｈＡＳＭＣ基础的ＤＮＡ合成（ｃｐｍ值为：１０３８５±３２６３ｖｓ，２５５４１±６４２１，Ｐ＜０．０１）及外源性ＰＤＧＦ诱导的ＤＮＡ合成（ｃｐｍ值为：９８７８±１９４７ｖｓ．１３４８１±４４ｌ０，Ｐ＜０．０５）；抑制ＰＤＧＦＡ链、ＴＧＦ－Ｂｐ和ＥＴ－１ｍＲＮＡ表达，提高ＰＤＧＦａ和β受体ｍＲＮＡ的表达；显著降低ｈＡＳＭＣ培养液中血管紧张素Ⅱ（ＡｎｇⅡ）的浓度和血管紧张素转换酶（ＡＣＥ）的活性，推测ＨＳＰＧ抑制ＰＤＧＦＡ链、ＴＧＦ－β及ＥＴ－１ｍＲＮＡ表达，降低ＡＣＥ活性及ＡｎｇⅡ浓度是其抑制ｈＡＳＭＣ增殖的重要机?

H-TdR incorporation,Northern blot analysis,reverse transcription-polymerase chain reaction(RT-PCR),radioimmunoassay(RIA) and colorimetric techniques were used to elucidate the effects of heparan sulfate proteoglycan(HSPG),extracted from healthy human aorta,on DNA synthesis and the gene expression of platelet-derived growth factor(PDGF),PDGF-receptors,transforming growth factor B(TGF-B),basic fibroblast growth factor(bFGF) and endothelin-1(ET-1) and also on renin-angiotensin system(RAS) in cultured human aortic smooth muscle cells(hASMC) and/or umbilical vein endothelial cells(hUVEC).The results showed that HSPG evidently inhibited the basic and PDGF-induced DNA synthesis of cultured hASMC(10385±3263 vs.25541±6421 cpm,P<0.01,and 9878±1947 vs.13481±4410 cpm,P<0.05,respectively);markedly inhibited the gene expression of PDGF A chain,TGF-βand ET-1,but stimulated receptors for PDGF;and significantly decreased the concentrarion of angiotensin Ⅱ and the activity of angiotensin converting enzyme in the media.It is suggested that the inhibitory effects of HSPG on gene expression of PDGFA and TGF-B in hASMC and that of ET-1 in hUVEC,together with the inhibitory effect on RAS might be the important mechanisms by which HSPG inhibits the proliferation of aortic smooth muscle cells. Furthermore,both HSPG and bFGF stimulated the proliferation of hUVEC with the rate of 37% and 95%,respectively.HSPG,together with bFGF.in the same dosage as used separately,showed a synergetic action on hUVEC growth.Although HSPG itself had no effect on the expression of bFGF mRNA,the stimulatory action of it on hUVEC could be neutralized completely by anti-bFGF antibody.This finding suggests that the stimulation of hUVEC by HSPG might proceed through the activation of endogenous bFGF.