细胞因子对体外培养的人球后成纤维细胞增殖及合成透明质酸,胶？…

目的 探讨甲状腺相关眼病（ＴＡＯ）的细胞免疫机制，并为临床上开展细胞因子或抗细胞因子疗法提供理论依据。方法 采用Ｔｈ１细胞代表细胞因子干扰素（ＩＦＮ）－γ和Ｔｈ２细胞代表细胞因子白细胞介素（ＩＬ）－４，刺激体外培养的人球后成纤维细胞（ＲＦ），并用液体闪烁测量和放射身显影技术观察ＲＦ的增殖和合成透明质酸（ＨＡ）、Ⅳ型胶原蛋白的能力。结果 ＩＦＮ－γ明显刺激ＦＲ增殖及合成ＨＡ，抑制其合成Ⅳ型胶原蛋白；     

去甲肾上腺素,人参皂甙和硝苯啶对大鼠动脉平滑肌细胞增殖和D…

目的 血管的老化与血管平滑肌细胞（ＶＳＭＣ）的异常增殖有关，我们拟探讨其增殖机制并寻求有效防治措施。 方法 应用噻唑蓝（ＭＴＴ）染色法和＾３Ｈ－ＴｄＲ掺入法观察去甲肾上腺素（ＮＥ）、硝苯啶（Ｎｉｆ）以及人参皂甙（Ｇｉｎ）对老年（１８月龄）和青年（３月龄）大鼠培养的主动脉平滑肌细胞（ＡＳＭＣ）增殖和细胞内ＤＮＡ合成的影响。 结果 （０．１￣５）×１０＾－５ｍｏｌ／Ｌ的ＮＥ可明显促进ＡＳＭＣ呈剂量依赖     

硝苯啶抗动脉粥样硬化作用的研究进展

硝苯啶抗动脉粥样硬化作用的研究进展山东省医学科学院林宪如综述张善同审校硝苯啶（Ｎｉｆｅｄｉｐｉｎｅ，Ｎｉｆ），是临床上常用的第一代钙拮抗剂（ＣａｌｃｉｕｍＡｎｔａｇｏｎｉｓｔｓ）药物。自美国学者Ｈｅｎｒｙ和Ｂｅｎｔｌｅｙ［１］首次报道Ｎｉｆ对实验性动...     

辛伐他汀对大鼠肺成纤维细胞增殖和胶原合成的影响及意义

目的探讨辛伐他汀（SIM）对肺成纤维细胞（LF）的增殖、胶原合成及基质金属蛋白酶-2（MMP-2）分泌的影响。方法用消化法培养新生SD大鼠的LF，给予不同浓度的SIM干预。四氮唑蓝（MTT）比色法检测细胞增殖，细胞免疫组化法测定细胞胶原的合成，ELISA法测定细胞培养上清液MMP-2的含量。结果随着SIM浓度（5，10，50μmol／L）的增高，MTT比色法A490值、LF表达Ⅰ、Ⅲ型胶原的平均光密度值（A）及培养上清液中的MMP-2含量均呈递减的趋势，与对照组相比，差异均非常显著（均P〈0．01）。加入200μmol／L甲羟戊酸可明显拮抗此作用（P〈0．01）。均具有统计学意义。结论降脂药物SIM具有抑制基本的成纤维细胞生长因子促LF增殖和胶原合成的作用，并可减少MMP-2的分泌，抑制LF黏附迁移功能，可能对防止肺移植术后闭塞性细支气管炎有一定的潜在临床应用价值。     

槲皮素对人成纤维细胞增殖及胶原合成的影响

目的探讨槲皮素对人成纤维细胞(HSF)生长及胶原分泌能力的影响。方法原代培养HSF,并以不同浓度的槲皮素干预24 h,以MTT法检测细胞增殖能力,以流式细胞术测定其细胞周期,对培养上清用碱水解法测定其中羟脯氨酸(Hyp)含量,酶联免疫吸附试验(ELISA)法测定Ⅰ型胶原(ColⅠ)及透明质酸(HA)含量。结果槲皮素作用HSF 24 h后,与对照组比较,槲皮素各剂量组细胞处于增殖期的细胞数目、增殖能力逐渐增加(P<0.05),并呈现一定的剂量效应关系;不同浓度槲皮素作用于HSF 24 h后,培养上清中Hyp、ColⅠ及HA含量均明显高于对照组(P<0.05),并具有剂量-效应关系。结论槲皮素可促进HSF增殖、刺激其分泌胶原及HA,对维持皮肤弹性、减少皮肤皱纹形成、延缓衰老有重要意义。     

糜酶对大鼠心脏成纤维细胞增殖和胶原合成的影响

目的探讨糜酶对大鼠心脏成纤维细胞（CFs）增殖和胶原合成的影响。方法用胰酶消化法分离、培养新生SD大鼠的CFs,采用MTT比色法（A490值）测定细胞数目,流式细胞术分析细胞周期,3H-脯氨酸掺入法测定总胶原合成,RT-PCR检测Ⅰ型和Ⅲ型胶原mRNA的表达。结果①糜酶以剂量依赖方式增加CFs的数目。其中15、30和60μg/L组的A490值分别为0.263±0.033、0.348±0.031和0.387±0.026,均高于对照组的A490值（0.201±0.019）,并有非常显著性差异（P<0.01）。②随着糜酶作用浓度的增加,CFs在G0/G1期的百分率逐渐减少,S期的百分率和细胞增殖指数逐渐增加。其中15、30和60μg/L组与对照组相比,上述各项指标均有显著或非常显著性差异（P<0.05或P<0.01）。③糜酶以剂量依赖方式增加CFs的3H-脯氨酸掺入量。其中15、30和60μg/L组的3H-脯氨酸掺入量分别为（520±75）、（684±62）和（769±58）cpm/孔,均高于对照组[（435±60）cpm/孔],差异有显著或非常显著性意义（P<0.05或P<0.01）。④随着糜酶作用浓度的增加,CFs的Ⅰ型和Ⅲ型胶原mRNA表达水平均呈递增趋势。其中15、30和60μg/L组的Ⅰ、Ⅲ型胶原mRNA表达的水平均显著高于对照组（P<0.01）。结论糜酶具有促进CFs增殖和胶原合成的作用,提示其可能在心肌纤维化过程中发挥重要作用。     

Urocortin对心肌成纤维细胞增殖和胶原合成作用的影响

目的 :探讨 urocortin对大鼠心肌成纤维细胞 （CFs）增殖和胶原合成作用的影响。方法 :消化法培养新生Sprague-Dawley （SD）大鼠的 CFs,采用四氮唑盐 （MTT）比色法测定细胞数目 ,流式细胞分析仪 （FCM）检测细胞周期 ,EL ISA法检测 型胶原合成 ,分别观察不同浓度 urocortin对 CFs增殖和胶原合成的影响。结果 :1随着 uro-cortin浓度的增高 ,CFs MTT比色法 A490 值呈明显的递增趋势 ,其中 10 - 10 、10 - 9、10 - 8、10 - 7和 10 - 6 mol/L组的 A490值分别为 0 .183± 0 .0 0 4、0 .2 2 2± 0 .0 0 4、0 .2 51± 0 .0 0 7、0 .2 80± 0 .0 0 6和 0 .3 17± 0 .0 0 2 ,均较对照组 （A490 值 0 .167± 0 .0 0 4）显著升高 （P均 <0 .0 1）。 2 FCM细胞周期分析结果表明 ,10 - 7mol/L urocortin作用 48h,CFs的 G0 /G1期细胞百分率均较对照组显著降低 （P<0 .0 1） ,S期细胞百分率、G2 /M期细胞百分率和增殖指数则较对照组显著增高 （均 P<0 .0 1）。 3 CFs的 型胶原分泌随着 urocortin作用浓度的增高呈明显的递增趋势 ,其中 10 - 10、10 - 9、10 - 8、10 - 7和 10 - 6 m ol/L urocortin组的 型胶原 A值分别为 0 .576± 0 .0 10、0 .614± 0 .0 0 8、0 .771± 0 .0 47、0 .83 9±0 .0 54和 0 .90 8± 0 .0 0 6,均较对照组     

毫米波辐照对心肌成纤维细胞增殖及胶原合成的影响

目的观察毫米波辐照对心肌成纤维细胞增殖及胶原合成功能的影响。方法取原代培养的心肌成纤维细胞,随机分为A、B、C、D组。A组为对照组。B、C、D组分别给予波长8.4 mm（频率35.75 GHz）、功率10 mW/cm^2的毫米波辐照15、30、60 min,1次/d,共5 d。辐照2、5 d用MTT法检测各组细胞增殖活性,辐照1、2、3、4、5 d分别用羟脯氨酸碱水解法和放射免疫分析法分别检测各组细胞培养液上清中的羟脯氨酸（HYP）和Ⅲ型前胶原N端肽（PⅢNP）。结果与A组相比B、C、D组CFbs增殖活性升高,细胞培养液上清中HYP和PⅢNP水平升高（P均〈0.05）,且升高程度均随毫米波每次照射时间及照射天数增加而增加（P均〈0.05）。结论毫米波辐照可以促进成纤维细胞的增殖及胶原蛋白合成。用毫米波辐照治疗心血管疾病时,每次辐照时间及疗程较长有导致心肌纤维化的可能性。     

The effect of rheumatoid synovial fluid macrophages on DNA,glycosaminoglycan and collagen synthesis by synovial fibroblasts

Summary The effects of soluble factors secreted by peripheral blood monocytes and rheumatoid synovial fluid macrophages were tested on human synovial fibroblast cultures. Both monocytes and macrophages liberated factors which reduced DNA synthesis (³H-thymidine incorporation) by synovial fibroblasts. Monocyte and macrophage factors stimulated hyaluronic acid synthesis. The activation obtained with rheumatoid synovial macrophages was considerably greater than that with monocytes. Foetal bovine serum was found to have a clear stimulatory effect on the synthesis of collagen and other proteins by fibroblasts. The effects of monocyte and macrophage factors on protein synthesis in synovial fibroblasts were small: collagen synthesis was slightly increased relative to other extracellular proteins.     

Altered distribution of collagen type I and hyaluronic acid in the cardiac outflow tract of mouse embryos destined to develop transposition of the great arteries

Summary Complete transposition of the great arteries (TGA) is inducible by treatment with all-trans retinoic acid in the ICR mouse. In this model, hypoplasia and dysplasia of the proximal outflow tract cushion tissue lead to non-spiral septation. In order to evaluate the effect of retinoic acid on the extracellular matrix of the cardiac outflow tract, we examined the distribution of collagen type I and hyaluronic acid, immunohistochemically, on days 8–9 of gestation. In controls, collagen type I fibrils ran mainly in a radial direction, extending towards the endocardium in the cardiac jelly of the proximal outflow tract. Also, a pair of longitudinal fiber bundles were formed stretching to the distal outflow tract. As for hyaluronic acid, intense staining was observed in the submyocardial and intermyocardial space of the outer curvature of the heart. On the other hand, in retinoic acid-treated embryos, the submyocardial radial fibrils or longitudinal fiber bundles of collagen type I were diminished, and irregular and dense deposits of collagen type I were observed along the endocardium. Furthermore, hyaluronic acid showed a loss of differential localization between the outer and inner curvature. Instead, irregular and intense staining was observed uniformly along the outflow myocardium. Thus, retinoic acid appeared to have perturbed the differentiation in the proximal outflow tract causing an altered organization of multiple extracellular matrix molecules, including collagen type I and hyaluronic acid, which led to an abnormal molecular network of the cardiac jelly in the cardiac outflow tract, abnormal septation and, further, to TGA or TGA-type anomalies.     

透明质酸对体外培养大骨节病软骨细胞增殖与凋亡的影响

目的 通过观察透明质酸(HA)对体外培养的大骨节病(Kashin-Beck disease,KBD)软骨细胞增殖、凋亡的影响,为临床上HA治疗KBD提供实验依据.方法 依据<大骨节病诊断标准>(GB 16003-1995)收集KBD患者和遭遇意外事故的病人(对照组)关节软骨,分离、体外培养关节软骨细胞.选用第2代细胞进行实验.两组软骨细胞分别给予不同剂量的HA,按HA剂量分为0、100、500 mg/L组.通过二苯甲唑溴盐(MTT)实验,测定第2、4、6天HA对KBD组、对照组软骨细胞增殖的影响.并通过流式细胞检测观察HA对软骨细胞凋亡的影响.结果 对照组在第4天时,500 mg/L组(0.140 ±0.049)促软骨细胞增殖作用大于0 mg/L组(0.116±0.021);KBD组在第6天时,500 mg/L组(0.179±0.081)与0 mg/L组(0.128 ±0.017)比较,显示了明显的促增殖作用(P<0.05).KBD组细胞凋亡率100、500 mg/L组(10.458±1.143、7.877±1.346)均较0 mg/L组(12.860±2.159)下降(P<0.05);对照组500 ms/L组(4.045±1.204)较0 mg/L组(7.128±1.244)细胞凋亡率下降(P<0.05).结论 HA对KBD软骨细胞具有促进增殖和抑制软骨细胞凋亡的作用,其中500 mg/L的HA改善KBD软骨细胞代谢的作用较100 mg/L明显.     

乙酰半胱氨酸对小鼠肺成纤维细胞系3T3增殖及胶原合成的影响

目的 研究乙酰半胱氨酸(NAC)对小鼠肺成纤维细胞3T3增殖、凋亡和胶原合成的影响.方法 培养小鼠肺成纤维细胞3T3,不同浓度NAC处理后,MTT检测细胞增殖,流式细胞仪检测细胞凋亡率、细胞周期,RT-PCR检测Ⅰ型前胶原mRNA表达的变化.结果 ①5、10、20、40 mmol/L NAC对3T3细胞生长均有明显抑制作用,且呈剂量依赖性;②5、10、20、40 mmol/L NAC作用24 h后,细胞凋亡率分别为(27.64±17.22)%、(33.73±14.82)%、(42.30±9.81)%和(69.27±11.50)%,均高于对照组(3.48±1.13)%(P<0.05或P<0.01);③5、10、20、40 mmol/L NAC能够引起G0/G1期细胞比例明显升高,S期比例明显降低(P值均<0.01);④5、10、20、40mmol/L NAC处理组3T3细胞Ⅰ型前胶原mRNA表达均低于对照组(P<0.05或P<0.01).结论 NAC可直接抑制小鼠肺成纤维细胞增殖,诱导其发生凋亡,并降低其胶原合成.     

The effects of synovial fluid macrophages and interleukin-1 on hyaluronic acid synthesis by normal synovial fibroblasts

Summary The effects of peripheral blood monocyte and rheumatoid synovial fluid macrophage conditioned media were studied on hyaluronic acid (HA) metabolism of normal synovial fibroblasts. Both media stimulated HA synthesis about two-fold compared to controls (1% fetal calf serum). The activated mononuclear phagocyte conditioned media did not contain HA-degrading activity in these experiments. The effects of various concentrations of interleukin-1 (IL-1) on HA synthesis and proliferation of synovial fibroblasts were studied. Even at very low concentrations (0.1 IU IL-1/ml) HA synthesis was stimulated. With increasing concentrations HA synthesis did not increase but proliferation was stimulated. Stimulated fibroblasts synthesized mainly high molecular weight HA. Thus with IL-1-activation, normal synovial fibroblasts could not produce increased amounts of abnormal HA with decreased molecular weight.     

Effects of different hyaluronic acid products on synovial fluid NO levels in knee osteoarthritis

The aim of the present study was to evaluate the effects of different hyaluronic acid forms on synovial fluid nitric oxide (NO) levels and glutathione peroxidase (GSHPx) activities in the treatment of patients with knee osteoarthritis (OA). Forty patients were equally randomized into two groups and treated with native sodium hyaluronate (group I) or with cross-linked hylan G-F 20 (group II). Clinical evaluations and synovial fluid aspirations were performed before the first (baseline), the second (week 1), and third injections (week 2), and a week after the third injection (week 3). NO levels were reduced at the end of the study in both groups (p<0.01 in group I, p=0.001 in group II), while no significant change was found in GSHPx activity. Also, the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) pain scores and WOMAC physical function scores were gradually improved at all follow-ups in the two groups. No significant differences between the two groups of NO levels, GSHPx activity, WOMAC pain scores, WOMAC stiffness scores, and WOMAC physical function scores were recorded during the study. Intra-articular hyaluronic acid therapy may reduce synovial fluid NO levels. These effects do not seem to be dependent on the molecular weight and various structural changes of hyaluronan products.     

抗坏血酸和透明质酸对木瓜酶致软骨损伤防护作用的研究

本文以人工流产的人胚四肢软骨作材料，经组织培养．通过电镜观察抗坏血酸和透明质酸对活化本瓜酶所致软骨溶解、破坏的防护作用．结果看到这两种物质对活化木瓜酶所致软骨损伤都有明显的防护作用．     

腐殖酸对鼠胶原脯氨酸及赖氨酸羟化的影响

目的探讨腐殖酸对NMRI鼠皮肤、肋骨及骨胶原合成,胶原分子中脯氨酸及赖氨酸羟化程度的影响.方法用含300×10-6腐殖酸的饮水饲养二代NMRI鼠,应用生物化学及分子生物学方法对子代鼠皮肤、肋骨及骨组织中胶原含量、脯氨酸及赖氨酸羟化程度进行观察.结果与对照组相比,实验组皮肤、肋骨及骨中胶原含量未见明显变化,胶原分子中脯氨酸羟化程度未见明显差异,而赖氨酸羟化程度则有明显增高(P＜0.05).结论胶原赖氨酸羟化程度的增高与腐殖酸对胶原合成过程中细胞内转录后修饰过程的调节作用有关.     

胃癌患者血清透明质酸的变化

目的：研究胃癌患者血清透明质酸的变化。方法：采用ＲＩＡ法测定胃癌患者４８例，胃良性病变患者４０例和健康人４０例，血清透明质酸水平。结果：胃癌患者血清ＨＡ水平（１３５．４±９２．６μｇ／Ｌ）明显高于胃良性病变患者和健康人（８０．７±２８．４，μｇ／Ｌ，７９．６±２１．３μｇ／Ｌ，Ｐ＜０．０１）。结论：透明质酸可反应胃癌的生物学特征，并用于胃癌的诊断