渔区年轻人主动脉蛋白聚糖和嗜苏丹病变的定量分析

本文对渔区３６例意外死亡的年轻人（１５～３９岁）新鲜（死后２４ｈ内）主动脉进行了蛋白聚糖和嗜苏丹病变定量分析，同时与以往报道的南宁和北京人主动脉的蛋白聚糖和嗜苏丹病变测定结果进行比较。结果是：①蛋白聚糖总量２１１４±５８４（μｇ糖醛酸／ｇ内中膜湿重，下同）；硫酸乙酰肝素－蛋白聚糖４８１±１２７，占蛋白聚糖总量的２３．１％，硫酸软骨素－蛋白聚糖１００４±３８６，占４６．７％；硫酸皮肤素－硫酸软骨素－蛋白聚糖６２８±１９５，占３０．２％；（２）主动脉内膜嗜苏丹病变测试结果明显好于南宁，更好于北京。     

牛主动脉蛋白聚糖对培养的人脐静脉内皮细胞生长的影响

为观察牛主动脉中蛋白聚糖对培养的人脐静脉内皮细胞生长的影响及其在动脉粥样硬化形成中的作用，用解聚提取，离子交换及凝胶过滤柱层析法从牛主动脉内，中膜分离出硫酸乙酰肝素蛋白聚糖、硫酸软骨素蛋白聚糖和硫酸皮肤素－硫酸软骨素蛋白聚糖。     

肝素及小肝素分子对培养平滑肌细胞的双重调节作用

为研究肝素及小肝素分子对培养的平滑肌细胞生长的作用,以探讨其是否可以作为一种抗血管平滑肌细胞增殖的药物,采用＾３Ｈ－胸腺嘧啶脱氧核苷掺入法观察肝素及小肝素分子对培养的人主动脉平滑机细胞合成ＤＮＡ的影响。肝素及小肝素分子的量以糖醛酸表示,以其浓度计算（１８、３５一７０ｍｇ／Ｌ）。结果发现,肝素及小肝素分子对生长良好的平滑肌细胞有抑制作用,不同浓度肝素的抑制率分别为６６％、６９％和６９％,而不同浓度小     

人主动脉硫酸乙酰肝素蛋白聚糖对胎儿主动脉平滑肌细胞c-fos和c-myc基因表达的影响

本室以往的研究发现硫酸乙酰肝素蛋白聚糖对培养的胎儿主动脉平滑肌细胞的增殖有明显的抑制作用。本实验采用Ｎｏｒｔｈｅｒｎｂｌｏｔ方法观察硫酸乙酰肝素蛋白聚糖对胎儿主动脉平滑肌细胞的ｃ－ｆｏｓ，ｃ－ｍｙｃ原癌基因的表达的影响，发现前者能抑制后者的表达。因此硫酸乙酰肝素蛋白聚糖抑制平滑肌细胞增殖的途径之一可能为抑制原癌基因的表达。     

人主动脉硫酸乙酰肝素蛋白聚糖对胎儿主动脉平滑肌细胞c-fos和c-myc基因表达的影响

本室以往的研究发现硫酸乙酰肝素蛋白聚糖对培养的胎儿主动脉平滑肌细胞的增殖有明显的抑制作用。本实验采用Ｎｏｒｔｈｅｒｎｂｌｏｔ的方法观察硫酸乙酰肝素蛋白聚糖对胎儿主动脉平滑肌细胞的ｃ－ｆｏｓ，ｃ－ｍｙｃ原癌基因的表达的影响，发现前者能抑制后者的表达。因此硫酸乙酰肝素蛋白聚糖抑制平滑肌细胞增殖的途径之一可能为抑制原癌基因的表达。     

人主动脉硫酸乙酰肝素蛋白聚糖对培养的胎儿主动脉平滑肌细胞c－fos基因mRNA表达的影响

人主动脉硫酸乙酰肝素蛋白聚糖对培养的胎儿主动脉平滑肌细胞ｃ－ｆｏｓ基因ｍＲＮＡ表达的影响刘俊明，丛祥凤，张春玲，张英珊（中国医学科学院心血管病研究所，阜外医院，北京１０００３７）人主动脉壁中的蛋白聚糖与动脉粥样硬化斑块的形成密切相关，而平滑肌细胞（Ｓ...     

N91对局部脑血流的影响

硫酸乙酰肝素蛋白聚糖对培养的人主动脉平滑肌细胞血小板源生长因子和肾素－血管紧张素系统的影响杨国君，张琪，丁金凤，孙巨忠（阜外医院生物化学研究室，北京，１０００３７）已知动脉平滑肌细胞增殖是动脉粥样硬化（Ａｓ）发病的重要因素，且已经证明硫酸乙酰肝素蛋白...     

碱性成纤维细胞生长因子反义寡脱氧核苷酸抑制胰岛素诱导动脉平滑肌细胞增殖

为研究胰岛素诱导的动脉平滑肌细胞增殖作用和碱性成纤维细胞生长因子反义寡脱氧核苷对培养的Ｗｉｓｔａｒ大鼠主动脉平滑肌细胞生长的影响。采用胰岛素和碱性成纤维细胞生长因子反义寡脱氧核苷酸处理培养的Ｗｉｓｔａｒ大鼠主动脉平滑肌细胞,Ｎｏｒｔｈｅｎｂｌｏｔ检测碱性成纤维细胞生长因子基因ｍＲＮＡ表达,并测定氚标胸腺嘧啶脱氧核苷掺入和细胞计数。结果发现胰岛素能明显诱导平滑肌细胞碱性成纤维细胞生长因子ｍＲＮＡ表达和增殖,且呈浓度依赖性。胰岛素浓度由０．００１ｍｇ／Ｌ增加到１．０ｍｇ／Ｌ,平滑肌细胞增殖率由１０％增加到５３％。碱性成纤维细胞生长因子反义寡脱氧核苷酸（５μｍｏｌ／Ｌ）能明显抑制胰岛素诱导的平滑肌细胞碱性成纤维细胞生长因子ｍＲＮＡ表达和增殖,氚标胸腺嘧啶脱氧核苷掺入被抑制４１．１％（与对照组比较,Ｐ＜０．０１）,细胞计数被抑制３０．２％（与对照组比较,Ｐ＜０．０１）。提示胰岛素可通过诱导平滑肌细胞碱性成纤维细胞生长因子的表达而促进动脉平滑肌细胞增殖,碱性成纤维细胞生长因子反义寡脱氧核苷酸能有效抑制胰岛素诱导的平滑肌细胞的ＤＮＡ合成及其增殖。     

碱性成纤维细胞生长因子和白细胞介素6促进大鼠血管平滑肌细胞增殖的协同作用

为观察碱性成纤维细胞生长因子和白细胞介纱６促进血管平滑肌细胞增殖的协同作用,采用大鼠主动脉贴块法培养血管平滑肌细胞,用５－溴－２－脱氧尿嘧啶掺入法和细胞计数法观察碱性成纤维细胞生长因子和白细胞介素６对平滑机细胞增殖的影响。结果发现,０￣１０．０μｇ／Ｌ碱性成纤维细胞生长因子和０￣１００μｇ／Ｌ白细胞介素６能促进平滑肌细胞增殖,且呈剂量依赖性,１．０μｇ／Ｌ碱性成纤维细胞生长因子和１．０μｇ／Ｌ白细     

食鱼对年轻人主动脉壁的影响和动脉粥样硬化早期病变的研究

用国家“七五”课题方法，对５１例渔区（舟山、宁波）意外死亡年轻人（１５～３９岁）尸检主动脉动脉粥样硬化（ＡＳ）早期病变进行了研究。数据经统计学处理，并与国家“七五”课题资料结果相比较。显示：渔区人主动脉内膜嗜苏丹病变（ＳＬ）面积百分比明显小于南宁，更小于北京；胸主动脉壁蛋白聚糖（ＰＧ）总量渔区低于南宁（Ｐ＜０．０１），硫酸乙酰肝素－蛋白聚糖（ＨＳ－ＰＧ）相对百分含量略高于南宁（Ｐ＞０．０５），亦高于北京（Ｐ＜０．０５）；主动脉内膜平滑肌细胞核数密度渔区大于南宁（Ｐ＜０．０５），面密度小于北京（Ｐ＜０．０５），中膜平滑肌细胞核面密度明显小于南宁及北京（Ｐ＜０．０１）；渔区主动脉ＡＳ病变检出率：Ⅰ、Ⅱ级病变与南宁、北京无差异，Ⅲ、Ⅳ、Ⅴ级病变均明显轻于南宁和北京（Ｐ＜０．０１）。结果提示：多食海鱼的生活习惯是导致鱼区人ＡＳ群的病变程度轻的重要因素之一。     

人脐静脉内皮细胞条件培养液对离体平滑肌细胞增殖的影响

为了探讨培养汇合后的人脐静脉内皮细胞条件培养液对培养的人胎儿主动脉平滑肌细胞增殖的影响，在收集人静脉静脉内皮细胞条件培养液后，用ＤＥＡＥ－Ｓｅｐｈａｃｅｌ离子交换柱和不同浓度的ＮａＣｌ溶液梯度洗脱，分为Ａ液、Ｂ液和Ｃ液三种成分，用＾３Ｈ－ＴｄＲ掺入法及细胞计数法分别观察上述三种培养液对培养的人胎儿主动脉平滑肌细胞增殖的影响，发现三种培养液对培养的人胎儿主动脉平滑肌细胞的增殖均有抑制作用，抑制程度依     

Heparin stimulates proteoglycan synthesis by vascular smooth muscle cells while suppressing cellular proliferation.

We studied the effect of heparin on proteoglycan synthesis by bovine aortic smooth muscle cells in culture. Confluent, growth-arrested cells were incubated with [35S]sulfate, [3H]glucosamine or [3]serine in the presence of 0-600 micrograms/ml heparin. Metabolically labeled proteoglycans secreted into the culture medium and associated with the cell layer were analyzed. In cultures treated with heparin there was a dose-dependent increase in [35S]sulfate incorporation into secreted proteoglycans which reached a maximum (35% above controls) at 100 micrograms/ml heparin. At higher concentrations of heparin, the stimulatory activity declined and finally disappeared. Radioactivity in cell-associated proteoglycans increased significantly (16% above controls) only in cultures treated with 100 micrograms/ml heparin. Heparin also produced similar increases in the incorporation of [3H]glucosamine and [3H]serine into secreted and cell-associated proteoglycans. While chondroitin sulfate, dermatan sulfate and heparan sulfate were elevated in the media, only chondroitin sulfate and heparan sulfate were increased in the cell layer. Heparin did not alter the degradation of proteoglycans. Heparin, while inhibiting the proliferation of subconfluent smooth muscle cells, also stimulated to a greater extent the incorporation of [35S]sulfate into proteoglycans. Other glycosaminoglycans, such as heparan sulfate, dermatan sulfate, heparin hexasaccharide and Sulodexide caused a significant but lesser stimulation of proteoglycan synthesis, while chondroitin sulfates and hyaluronic acid had no effect. Gel filtration chromatography of proteoglycans and their constituent glycosaminoglycans from heparin-treated and untreated cultures showed no differences in their molecular size. The results indicate that heparin can stimulate proteoglycan synthesis by vascular smooth muscle cells irrespective of their state of proliferation. This might have implications in vessel wall repair and arterial wall lipid deposition.     

Carbon-13 nuclear magnetic resonance study of chondroitin 4-sulfate in the proteoglycan of bovine nasal cartilage.

The Fourier transform 13C nuclear magnetic resonance spectra of bovine nasal cartilage proteoglycan subunit and complex and whole bovine nasal cartilage were obtained and compared with that of chondroitin 4-sulfate. The spectrum of chondroitin 4-sulfate in solution revealed multiple resolvable resonances with linewidths that are consistent with considerable segmental motion in the polysaccharide chain. The spectra of proteoglycan subunit and complex in solution and that of whole cartilage were very similar to that of free chondroitin 4-sulfate chains. This indicates that the linkage of multiple chondroitin sulfate chains to proteoglycan core protein and the association of proteoglycan with collagen and other constituents of cartilage matrix does not significantly alter the structure and motions of these chains.     

Effect of growth factors on hyaluronan and proteoglycan synthesis by retroocular tissue fibroblasts of Graves' ophthalmopathy in culture.

One of the pathological changes seen in Graves' ophthalmopathy is the deposition of glycosaminoglycans such as hyaluronan and proteoglycan in retroocular connective tissue. We analyzed glycosaminoglycans synthesized by retroocular tissue fibroblasts in culture derived from an individual not suffering from thyroid disease and from three patients with Graves' ophthalmopathy. Retroocular tissue fibroblasts synthesized both hyaluronan and proteoglycan, the latter composed mainly of chondroitin sulfate. This contrasts with the proteoglycan synthesized by adult skin fibroblasts which was composed of dermatan sulfate and heparan sulfate proteoglycan. Chondroitin sulfate proteoglycan secreted by retroocular tissue fibroblasts consisted of large and small chondroitin sulfate proteoglycans (CS-PG), their size being determined by the Sepharose CL-6B column. The effects of IGF-1 and PDGF on hyaluronan and proteoglycan synthesis were studied separately and in combination. Both IGF-1 and PDGF increased the synthesis of hyaluronan and proteoglycan in a dose-dependent manner. IGF-1 predominantly stimulated secretion of small CS-PG, while PDGF increased large CS-PG markedly when studied in retroocular tissue fibroblasts. In contrast, IGF-1 stimulated secretion of small proteoglycan while PDGF had little effect on proteoglycan synthesis in skin fibroblasts. Thus, glycosaminoglycan synthesized by retroocular tissue fibroblasts has a unique composition and each component is regulated independently, at least in part.     

Proteoglycans and vascular cell proliferation

The relationships that exist between proliferative states and proteoglycan (PG) synthesis have been examined in monkey aortic smooth muscle cells in culture. These cells were made quiescent in medium containing low serum (0.1%) and stimulated to divide by addition of either nanogram quantities or platelet-derived growth factor (PDGF) or medium containing 5% serum. Incorporation of [35S]sulfate into PG was increased during the first 24 h of growth stimulation, and this increase appeared to be principally in the large chondroitin sulfate proteoglycan (CSPG). Furthermore, addition of p-nitrophenyl beta-D-xyloside, which perturbs PG metabolism, inhibits cells from proliferating, suggesting that PG may be involved in facilitating cell division. Inhibition of cell proliferation by heparin and/or TGF-beta also causes elevated levels of 35S-sulfate incorporation into PG by these cells. These studies indicate that proteoglycan metabolism is modulated as a function of the growth of arterial smooth muscle cells; however, it is still uncertain whether PG play a direct or indirect role in the control of cell growth.     

Low density lipoprotein binding affinity of arterial wall isomeric chondroitin sulfate proteoglycans

Although the selective interaction of low density lipoproteins (LDL) with arterial proteoglycans is known, information is lacking on LDL-binding affinity of different subspecies occurring within a proteoglycan family. Isomeric chondroitin sulfate proteoglycan preparations sedimenting at densities of 1.54 g/ml (D1), 1.50 g/ml (D2) and 1.46 g/ml (D3) were isolated from bovine aorta intima-media under dissociative conditions and subjected to equilibrium binding to LDL-agarose gel. D1, D2 and D3 contained 36%, 37% and 11% dermatan sulfate, respectively. Sulfate to hexosamine ratio was low (0.73) in D1 when compared to D2 and D3 (0.94 and 1.04). Of the total proteoglycans contained in D1, D2 and D3, 41%, 52% and 66% interacted with LDL, respectively. LDL-bound proteoglycans dissociated over a wide range of ionic strengths (0.15-1.0); in comparison, LDL-bound heparin dissociated within a narrow range (0.5-0.75). Unlike other preparations, 30% of bound D3 dissociated at an ionic strength of 1.0. In D1 and D2 the proportion of dermatan sulfate increased in proteoglycan fractions that were bound firmly to LDL, whereas a high affinity fraction in D3 contained no dermatan sulfate. Thus, isomeric chondroitin sulfate proteoglycans display considerable divergence with respect to LDL binding. This may depend not only on the degree of sulfation but on other characteristics of the chondroitin sulfate isomers as well.     

同型半胱氨酸对血管内皮细胞合成蛋白聚糖的影响

为探讨同型半胱氨酸对血管内皮细胞合成蛋白聚糖的影响，采用400umol／L同型半胱氨酸作用于培养的人脐静脉内皮细胞，以^35S-Na2SO4为示踪物标记细胞合成的蛋白聚糖，通过离子交换层析，凝胶过滤层析分离蛋白聚糖。结果发现，实验组培养液中总蛋白聚糖降低，硫酸乙酰肝素蛋白聚糖及硫酸软骨素-硫酸皮肤素蛋白聚糖含量也降低，但其百分含量未见改变。细胞层中蛋白聚糖未见明显变化。     

Characterization of proteoglycans synthesized by fetal rat lung type II pneumonocytes in vitro and the effects of cortisol

The synthesis of proteoglycans by primary cultures of 19-day gestation fetal rat lung Type II pneumonocytes was studied. The cells were grown in the presence of [3H]-glucosamine and/or [35S]-Na2SO4 and the radioactive label incorporated into proteoglycans was analyzed. Proteoglycans of high molecular weight (approximately 200 Kd) were isolated by gel permeation chromatography and contained both [3H] and [35S]. The glycosaminoglycan composition of the proteoglycans was determined by electrophoresis and autoradiography. The medium contained 65-80% of the labeled proteoglycans and was enriched for hyaluronate, with lesser amounts of the sulfated glycosaminoglycans (dermatan sulfate greater than heparan sulfate greater than chondroitin sulfate). The cell layers retained 20-35% of the labeled proteoglycans and was enriched for heparan sulfate, with lesser amounts of chondroitan sulfate greater than dermatan sulfate greater than hyaluronate. The synthesis of proteoglycans was time-dependent and was stimulated by increasing concentrations of fetal bovine serum. Cortisol inhibited proteoglycan synthesis, apparently by decreasing the availability of proteoglycan core-protein.