人主动脉壁硫酸乙酰肝素蛋白聚糖对培养的人主动脉平滑机细胞合成蛋白聚糖的影响

Effect of Human Aortic Heparan Sulfate Proteoglycan (HSPG) on Proteoglycan Synthesis of Cultured Human Aortic Smooth Muscle Cells (HASMC)

从人正常胸主动脉分离硫酸乙酰肝素蛋白聚糖（ＨＳＰＧ），观察其对体外培养的人主动脉平滑肌细胞（ＨＡＳＭＣ）合成ＰＧ的影响。ＨＡＳＭＣ在不加（对照）或加ＨＳＰＧ（１９μｇ醛酸／ｍｌ）的￣（３５）Ｓ－硫酸钠培养液中培养，以标记ＰＧ。继之，培养液及细胞层的４ｍｏｌ／Ｌ盐酸胍提取液中的ＰＧｓ经离子交换及凝胶过滤柱层析分离，发现加ＨＳＰＧ后，培养液中的ＨＳＰＧ，硫酸软骨素ＰＧ（ＣＳＰＧ）及硫酸皮肤素－硫酸软骨素ＰＧ（ＤＳＣＳＰＧ）均明显增高，而细胞层中仅ＨＳＰＧ和ＣＳＰＧ增高，且加ＨＳＰＧ后细胞层的ＤＳＣＳＰＧ分子大小有所不同，进一步分析ＤＳＣＳＰＧ中ＤＳ及ＣＳ含量发现加ＨＳＰＧ组ＨＡＳＭＣ细胞层中的ＤＳ％含量略低于对照组。结果提示ＨＳＰＧ可刺激ＨＡＳＭＣ的ＰＧ合成，其可能与血管壁修复及动脉壁脂质沉积有关。

For studying the effect of arterial HSPG on the synthesis of proteoglycans of cultured smooth muscle cells (SMC), human aortic HSPG and cultured human fetal aortic SMC (HASMC) were prepared. Cultured HASMC were firstly inhibited at G_0/G_1 by DMEM containing 0.2 % human serum (HS) and 0.2% FCS, and then incubated with DMEM containing 5% HS, 5% FCSand ￣(35)S-Na_2SO_4(50μCi/ml) with HSPG(19 μg uronic acid/ml) as the experimental group (E),and without HSPG as the control(C). The ￣(35)S-labelled PGs in both the culture medium and cell layer were extracted and isolated by Sephadex G-50, DEAE-Sephacel and Sepharose CL-4B chromatography. The glycosaminoglycans(GAGs) released by alkaline borohydride were analyzed by Sepharose 6B and characterized by chondroitinase AC-Ⅱand ABC digestion. In group E, dpm of ￣(35)S-HSPG, -CSPG and -DSCSPG in the media, dpm of ￣(35)S-HSPG, -CSPG in the cell layer were higher than those in the corresponding controls. Furthermore, the molecular size of DSCSPG from cell layer of group E was larger than that of group C with K_d of 0. 45 and 0. 56, respectively, together with slightly lower percentage of DS. It is suggested that human aortic HSPG could increase PG synthesis of cultured HASMC.