黄颜木素对HSC—T6细胞增殖和胶原合成的影响

目的：研究黄颜木素对激活的永生型大鼠肝腑 脂细胞－－ＨＳＣ－Ｔ６细胞增殖和胶原合成的影响。方法：细胞增殖采用结晶紫染色法检测,胶原合成采用＾３Ｈ－酸掺入法分析。结果：黄颜木素（６．２５－５０．００μｍｏｌ／Ｌ）以剂量依赖方式显著抑制血小板源生长因子（ＰＤＧＦ）刺激的ＨＳＣ－Ｔ６细胞增殖,并抑制生长因子因子β１（ＴＧＦβ１）诱导的细胞内胶原合成,结论：黄颜木还给具有抑制激活的肝储脂细胞增殖和胶原合成     

硝苯吡啶和维拉帕米治疗肝纤维化细胞学研究

目的研究硝苯吡啶（ｎｉｈｅｄｉｐｉｎｅｃ，Ｎｉｆ）和维拉帕米（ｖｅｒａｐａｍｉｌ，Ｖｅｒ）对人胚肺成纤维细胞（ｈｕｍａｎｌｕｎｇｆｉｂｒｏｂｌａｓｔｓ，ＨＬＦ）的ＤＮＡ和胶原合成的影响，从细胞水平评价钙拮抗剂防治器官纤维化并提供实验依据。方法采用ＭＴＴ比色法、３Ｈ－ＴｄＲ和３Ｈ－脯氨酸掺入法测定细胞增殖、ＤＮＡ合成和胶原合成。结果Ｎｉｆ和Ｖｅｒ在０～５０μｍｏｌ／Ｌ浓度范围内均以浓度依赖方式抑制ＨＬＦ增殖、ＤＮＡ合成及胶原合成（Ｐ＜０．０５～０．０１）。结论Ｎｉｆ和Ｖｅｒ具有抗纤维化作用     

粉防己碱对肺动脉高压大鼠的肺小动脉平滑肌细胞PDGFR—α,β表？…

本实验用免疫组织化学检测粉防己碱（Ｔｅｔ）对肺动脉高压大鼠肺小动脉中膜平滑肌细胞血小板生长因子受体α,β（ＰＤＧＦＲ－α,ＰＤＧＦＲ－β）表达的影响。结果表明,Ｔｅｔ能抑制肺小动脉中膜平滑肌细胞增殖时的ＰＤＧＦＲ－α,β的表达增强（Ｐ〈０．０１）,实验结果提示,Ｔｅｔ抑制ＰＤＧＦＲ－α,β的表达是其抗血管平滑肌细胞增殖,抑制肺动脉高压血管的构型重建的分子机制之一。     

pcDNA3-hbFGF转染角朊细胞对真皮成纤维细胞增殖的生物学效应

目的：观察含外源性人碱性成纤维细胞生长因子（ｈｂＦＧＦ）基因的角朊细胞（ＨＫ）,对真皮成纤维细胞（ＨＤＦｉｂ）增殖的影响;探讨转ｈｂＦＧＦ基因ＨＫ对创面愈合的生物学效应及作用机理,为这种转基因ＨＫ细胞用于创面基因治疗的可行性提供实验依据。方法：收集含真核表达质粒ｐｃＤＮＡ３－ｈｂＦＧＦ的ＨＫ细胞培养上清,加入培养的ＨＤＦｉｂ细胞内;ＭＴＴ法测定细胞的增殖率、＾３Ｈ－ＴｄＲ掺入法测定细胞ＤＮＡ合成率     

牙龈成纤维细胞与牙周韧带细胞在二种玻璃离子水门汀上附着及增？…

目的 比较人类牙龈成纤维细胞（ＨＧＦ）与牙周韧带细胞（ＰＤＬＣ）在两种玻璃离子水门汀（ＧＩＣ）上的附着及增殖情况。方法 利用体外细胞培养技术及＾３Ｈ－ＴｄＲ掺入法检测ＨＧＦ及ＰＤＬＣ在两种ＧＩＣ上的附着及ＤＮＡ合成。结果 ＨＧＦ与ＰＤＬＣ在培养板上的附着及ＤＮＡ合成差异无显著性（Ｐ〉０．０５）,但ＨＧＦ在ＧＩＣ上的附着及ＤＮＡ合成明显强于ＰＤＬＣ（Ｐ〈０．０５）。结论 Ｋｅｔａｃ Ｆｉｌ和Ｆｕｊｉ     

PDGF—B基因的膜型表达产物促进血管平滑肌细胞增殖和胶原合成

考察ＰＤＧＦ－Ｂ基因的膜型表达产物对血管平滑肌细胞（ＶＳＭＣｓ）增生、胶原合成的影响。制备供转导人ＰＤＧＦ－Ｂ基因的重组逆转录病毒，感染ＮＩＨ３Ｔ３细胞，受染ＮＩＨ３Ｔ３细胞经筛选、扩增后，制备细胞膜上表达产物ＰＤＧＦ－ＢＢ以观察其对ＶＳＭＣｓ生长的影响，并以３Ｈ－ＰｒｏＬｉｎｅ掺入率评估该重组蛋白对ＶＳＭＣｓ胶原合成的影响。结果：重组逆转录病毒介导ＰＤＧＦ－Ｂ基因转移并表达出具有生物学活性的重组ＰＤＧＦ－ＢＢ，免疫荧光细胞化学染色证实其表达；该膜型重组蛋白可促进ＶＳＭＣｓ增殖和胶原合成。     

IL－1α诱导牛脑微血管内皮细胞释放血小板衍生性生长因子

目的：研究白细胞介素－１α（ＩＬ－１α）诱导牛脑微血管内皮细胞（ＢＣＭＥＣ）释放血小板衍生性生长因子（ＰＤＧＦ），以及药物对由之引起的牛脑微血管平滑肌细胞（ＢＣＭＳＭＣ）增殖的拮抗作用。方法：体外培养ＢＣＭＥＣ和ＢＣＭＳＭＣ，结晶紫染色法测定细胞增殖。结果：ＩＬ－１α不能直接促进ＢＣＭＳＭＣ的增殖；但经ＩＬ－１α刺激的ＢＣＭＥＣ培养上清能显著地促进ＢＣＭＳＭＣ的增殖。这种增殖作用与ＩＬ－１α剂量呈正相关，并且可被抗ＰＤＧＦ抗体中和。欧芹素乙（Ｉｍｐ），异欧芹素乙（ｉｓｏ－Ｉｍｐ），６－（α，α－ｐｈｅｎｙ－ｌａｃｅｔｙｌｐｉｐｅｒａｚｉｎｙｌ）ｐｈｅｎｙｌ－５－ｍｅｔｈｙｌ－４，５－ｄｉｈｙｄｒｏ－３（２Ｈ）－ｐｙｒｉｄａｚｉｎｏｎｅ（ＰＭＤＰ）不影响ＩＬ－１α诱导ＢＣＭＥＣ释放ＰＤＧＦ，但对ＰＤＧＦ促ＢＣＭＳＭＣ增殖呈剂量依赖的拮抗。结论：ＩＬ－１α促进ＢＣＭＥＣ释放ＰＤＧＦ。ＩＬ－１α对ＢＣＭＳＭＣ增殖的促进作用需经ＰＤＧＦ等生长因子的介导。Ｉｍｐ，ｉｓｏ－Ｉｍｐ，ＰＭＤＰ拮抗ＰＤＧＦ引起的ＢＣＭＳＭＣ增殖。     

粉防己碱抑制大鼠肺动脉高压过程中PDGF-A、B表达的研究

目的：探讨血小板源生长因子（ＰＤＧＦ）在粉防己碱（Ｔｅｔ）抑制野百合碱（ＭＣＴ）诱导的大鼠肺动脉高压肺血管构形重建中的作用。方法：建立（ＭＣＴ）诱导的大鼠肺动脉高压模型,同进以两个实验组加不同剂量的Ｔｅｔ抑制肺动脉高压,用免疫组化方法检测Ｔｅｔ对ＭＣＴ致肺动脉高压在 肺小动脉中膜平滑肌细胞ＰＤＧＦ－Ａ、Ｂ表达的影响。结果：Ｔｅｔ能抑制肺小动脉中膜平滑肌细胞增殖时的ＰＤＧＦ－Ｂ的表达增强（Ｐ〈０．０     

重组生长激素对烫伤创面PDGF及其受体的影响

目的研究重组生长激素（ｒｈＧＨ）在促进创面愈合过程中对血小板源性生长因子（ＰＤＧＦ）及其受体的影响。方法采用大鼠深Ⅱ度烫伤模，观察大鼠深Ⅱ度烫伤后创面愈合过程中ｒｈＧＨ对ＰＤＧＦ、血小板源性生长因子受体（ＰＤＧＦＲ）、Ⅰ、Ⅲ型胶原、组织形态学和血胰岛素样生长因子Ⅰ（ＩＧＦ－Ⅰ）变化的影响。结果用ｒｈＧＨ治疗５、１０ｄ后，其创面ＰＤＧＦ、ＰＤＧＦ－α受体、ＰＤＧＦ－β受体的表达明显增强，Ⅰ、Ⅲ型胶原     

血小板源生长因子-AB对成纤维细胞周期及DNA合成的影响

目的：探讨血小板源生长因子（ＰＤＧＦ）－ＡＢ在创面愈合及瘢痕增生过程中的作用机制;方法：取第３代～６代体外培养的人正常皮肤成纤维细胞（ＮｓＦｂ）及增生性瘢痕成纤维细胞（ＨＴｓＦｂ）,经ＰＤＧＦ－ＡＢ作用后,用＾３Ｈ－ＴｄＲ参入法测定ＤＮＡ合成的变化,用汉式细胞术测定细胞周期的变化。结果：ＰＤＧＦ－ＡＢ作用后,两种细胞ＤＮＡ民均明显增加,Ｓ期细胞数百分比均明显增高,Ｇ１期细胞不断进入Ｓ期,但作用有划     

PDGF与肝素对动脉SMCI,Ⅲ型前胶原mRNA表达及胶原蛋白合成？…

目的 研究,探讨血小板源生长因子（ｐｌａｔｅｌｅｔｄｅｒｉｖｅｄｇｒｏｗｔｈｆａｃｔｏｒ,ＰＤＧＦ）和肝素对人主动脉平滑肌细胞（ｈｕｍａｎａｏｒｔａａｍｏｏｔｈｍｕｓｃｌｅ,ｃｅｌｌ,ｈＡＳＭＣ）增殖,胶原蛋白的合成,分泌以及Ｉ,Ⅲ型前胶原ｍＲＮＡ表达和转移生长因子β（ｔｒａｎｓｆｏｒｍｉｎｇｇｒｏｗｔｈｆａｃｔｏｒβ－ＴＧＦ－β）ｍＲＮＡ表达的调节作用,方法＾３Ｈ－ＴｄＲ,＾３Ｈ－脯氨酸掺入及Ｎ     

血小板衍生生长因子在梗阻性黄疸鼠肝中的免疫组化分析

通过大白鼠胆总管结扎模型，研究血小板衍生生长因子（ＰＤＧＦ）在肝内的免疫组化定位，探讨ＰＤＧＦ在梗阻性黄疸肝纤维化中的作用。实验结果：胆总管结扎２ｗ后肝小叶汇管区有明显的间质细胞和胆管增生及肝纤维化，胆总管结扎４ｗ导致肝硬变形成；肝脏ＰＤＧＦ－Ｂ链免疫组化染色显示胆总管结扎２ｗ后肝间质细胞和胆管上皮细胞呈阳性染色反应。表明ＰＤＧＦ在肝间质细胞增生、产生大量细胞外基质的过程中起重要作用，与胆道梗阻所致肝纤维化、肝硬变有关。     

Serum pharmacological effects of Fuzheng Huayu Decoction on Ito cell proliferation and collagen synthesis in rats) on Ito cell proliferation and collagen synthesis in rats

Objective: To explore mechanism of the effect of Fuzheng Huayu Decoction on Ito cell proliferation and collagen synthesis.Methods: Biological effects of the drug serum taken from rats fed with Fuzheng Huayu decoction (FZHYD) on cell proliferation and collagen synthesis of Ito cell line were observed. Serum collected from rats at various times (1, 2 or 3 hours) after animals were fed with FZHYD once (It) or twice (2t, 2nd time repeated with the same dose), was incubated with Ito cell.Results: The lt-2h serum in 0. 46 g/kg FZHYD group, it seems to prevent cell proliferation, but had no significant difference. While all 2t drug serum could inhibit cell proliferation. The 2t-lh drug serum had no obvious effects on Ito cell morphology, but could improve cell viability, its effect on cell proliferation was dose and serum concentration dependent. Also the 2t-lh drug serum in all doses could inhibit Ito cells both intracellular and extracellular collagen production, the drug serum of 0.46 g/kg group had inhibitory effect in serum concentration dependent manner and similar effect as colchicine.Conclusion: Inhibition of Ito cells proliferation and collagen synthesis is perhaps one of main mechanism of FZHYD in antifibrotic action.     

Studies on the inhibitory effect of tetrandrine on silica-induced fibrogenesis in vitro

Two cell culture systems composed of rat peritoneal macrophages and 2BS fibroblasts were used to study the inhibitory action of tetrandrine on collagen and glycosaminoglycan synthesis. The activation of silica-treated macrophage supernatants stimulated collagen synthesis by 2BS fibroblast cells. Tetrandrine had an inhibitory effect on the biosynthesis of collagen and glycosaminoglycans. The increase in the content of hydroxyproline, representing collagen, was verified by comparing it with that of DNA.     

Effects of ribozyme targeting platelet-derived growth factor receptor β subunit gene on the proliferation and apoptosis of hepatic stellate cells in vitro

Background Activation and proliferation of hepatic stellate cells (HSC) is essentially involved in the development and progression of hepatic fibrosis. The most potent growth factor for HSC is platelet-derived growth factor receptor (PDGF) and PDGF receptor β subunit (PDGFR-β) is the predominant signal transduction pathyway of PDGF which is overexpressed in activated HSC. This study investigated the cleavage activity of hammerhead ribozyme targeting PDGFR-β mRNA in HSC and the effect on biological characteristics of HSC.Methods Expression vector of anti-PDGFR-β ribozyme was constructed and transfected into rat activated HSC with lipofectamin. The positive cell clones were gained by G418 selection. The expression of PDGFR-β, α-smooth muscle actin, and typeⅠand type Ⅲ collagen were detected by using Northern blot, Western blot and immunocytochemical staining, respectively. The cell proliferation was determined with MTT colorimetric assay. The cell apoptosis was analyzed by using flow cytometry, acridine orange fluorescence vital staining and transmission electron microscopy.Results The expression of PDGFR-β at mRNA and protein level was markedly reduced in ribozyme-transfected HSC by 49%-57% (P<0.05-0.01). The proliferation and α-smooth muscle actin expression of ribozyme-transfected HSC were significantly decreased (P<0.05-0.01), and the type Ⅰ and type Ⅲ collagen synthesis were also reduced (P<0.01). In addition, the proliferative response of ribozyme-transfected HSC to PDGF BB was significantly inhibited. Otherwise, the apoptotic cells were significantly increased in ribozyme-transfected HSC (P<0.01), and typical apoptotic cells could be found under transmission electron microscopy.Conclusions The anti-PDGFR-β ribozyme effectively cleaved the target RNA and significantly inhibited its expression, which blocked the signal transduction of PDGF at receptor level, inhibited HSC proliferation and collagen synthesis, and induced HSC apoptosis. These results suggest that inhibiting PDGFR-β expression of HSC may be a new target for the therapy of liver fibrogenesis, and ribozyme may be a useful tool for inhibiting PDGFR-β expression.     

正常人皮肤成纤维细胞不同培养条件下的生物学特性

目的 观察成纤维细胞在不同培养条件下生长,增殖,代谢及蛋白质合成等情况,探讨成纤维细胞的最佳培养模型。方法 将正常皮肤中的成纤维细胞（NsFb）分离出来并进行原代及继代培养,之后建立不同的成纤维细胞培养系统,分别利用绘制细胞生长曲线,^3H-胸腺嘧啶（^3H-TdR）掺入及^3H-脯氨酸掺入等方法观察细胞的增殖,DNA代谢及胶原合成等情况,研究细胞在不同条件下的生物学特性。结果 成纤维细胞在单层及纤维蛋白胶中培养时细胞的增殖,代谢及蛋白质合成等生物学功能均明显强于两种胶原凝胶中培养时的情况。结论 以纤维蛋白胶为支架的三维培养系统能更好地模拟生理条件下创面愈合时成纤维细胞的生物学特性。     

联合应用粉防已碱与甘草酸抑制HSC增殖及细胞外基质合成

目的：探讨联合应用粉防已碱（tetrandrine,Tet）与甘草酸（glycyrrhizinic acid,Glz）对肝星状细胞增殖与细胞外基质合成的影响。方法：采用胶原酶二步原位灌注法分离、培养大鼠HSC，并分别给予2．5mg/L及5mg/L Tet、0．5mg/L Glz、2．5mg/L Tet 0.5mg/L Glz、5mg/L Tet 0.5mg Glz干预，分别检测各处理组HSC增殖及细胞外基质合成水平。结果：5mg/L Tet、0.5mg/L Glz和各浓度Tet联合Glz均能不同程度地抑制HSC 3H－TdR掺入和增殖（P＜0．05）；能够显著抑制体外培养HSC 3H－脯氨酸掺入（P＜0．05），降低上清液HA、Ln和CIV水平。5mg/L Tet联合Glz作用最为显著（P＜0．05）。结论：联合应用Tet与Glz能够更加有效地抑制HSC DNA合成、细胞增殖及ECM合成与分泌。     

肝纤维化过程中肝星状细胞的移行机制

目的探讨肝纤维化过程中肝星状细胞（HSC）移行的机制和肝纤维化病变过程中新的病理生理机制。方法运用改良的Boyden腔系统,在体外条件下模拟体内正常Disse间隙的微环境及肝纤维化时的相关改变,以HSC为研究对象,通过细胞迁移实验、明胶酶谱和凝胶免疫印迹等实验方法,研究肝纤维化时致纤维化生长因子和细胞外基质促进HSC移行的机制。结果肝纤维化时增高的血小板衍化生长因子BB（PDGF—BB）、转化生长因子β1（TGF—β1）以及上皮细胞生长因子（EGF）均可以刺激活化的HSC分泌基质金属蛋白酶2（MMP-2）,而MMP-2通过降解胶原又可以促进HSC的移行（4．9倍）;HSC的移行是由其表面的整合素α1和以所α2,不同致纤维化生长因子诱导HSC移行时依赖着不同的整合素的介导;由HSC分泌的细胞外基质对HSC自身的行为有反馈调节作用,间质类基质可促进HSC的移行（3．2倍）,而基底膜样的基质则可抑制HSC的移行（1．2倍）。结论肝纤维化时Disse间隙微环境的改变导致了HSC的移行,其机制与促进HSC高表达MMP-2相关;肝纤维化时HSC的移行由整合素α1和以所α2;不同的细胞外基质对HSC的移行行为有着不同的调节作用。     

粉防己碱对人肝癌7402细胞株增殖与凋亡的影响

目的：通过观察粉防己碱（Tetrandrine）对人肝癌7402细胞株增殖与凋亡的影响，初步探讨粉防己碱对肝癌的体外抗肿瘤效应。方法：采用四甲基偶氮唑蓝（MTT）比色法、集落形成实验观察粉防己碱对人肝癌7402细胞株增殖的抑制效应，利用细胞DNA琼脂糖凝胶电泳、细胞凋亡荧光染色法及流式细胞术检测粉防己碱诱导肿瘤细胞凋亡的作用，以免疫细胞化学法检测粉防己碱对Bcl-2和Bax蛋白表达水平的影响。结果：MTT比色法、集落形成实验显示，粉防己碱对人肝癌7402细胞株增殖有抑制作用，其抑制效应具有剂量依赖的特点，细胞凋亡荧光染色法、琼脂糖凝胶电泳及流式细胞术表明，粉防己碱可诱导7402细胞凋亡，免疫细胞化学法显示，粉防己碱上调Bax蛋白表达，下调Bcl-2蛋白表达。结论：粉防己碱对人肝癌7402细胞株增殖的抑制效应具有剂量依赖性，并可诱导细胞凋亡，其抗肿瘤效应可能与凋亡相关基因表达的调控有关。     

缓激肽对血小板源性生长因子诱导的大鼠血管平滑肌细胞增殖中一氧化氮及其合酶基因表达的影响祆

目的 探讨缓激肽在对血小板源性生长因子（ＰＤＧＦ）诱导的培养的大鼠胸主动脉血管平滑肌细胞（ＶＳＭＣｓ）增殖的影响中一氧化氮（ＮＯ）含量与诱导型一氧化氮合酶（ｉＮＯＳ）ｍＲＮＡ表达的变化。方法 取ＳＤ大鼠胸主动脉平没肌细胞进行培养,传至４～６代,分成以下几组：１．对照组;２．ＰＤＧＦ组（５０ｎｇ／ｍｌ）;３．ＰＤＧＦ＋缓激肽（１０＾－４ｍｏｌ／Ｌ组;４．ＰＤＧＦ＋缓激肽＋Ｈｏｅ１４０（１０＾－４ｍｏ