促长散对小鼠成纤维细胞生长的作用

目的观察促长散(促创伤愈合中药)对昆明小鼠成纤维细胞生长的影响。方法建立昆明小鼠的全层皮肤缺损创面模型,并随机分为3组,每组20只;大、小剂量实验组分别给予促长散172、374mg.kg-1,模型组给予0.9%生理盐水。10天后,取肉芽组织,分别在光镜、电镜下观察成纤维细胞和生长因子(bFGF)的表达。结果大、小剂量促长散实验组与模型组相比,显著提高了肉芽组织中成纤维细胞数和bFGF表达水平,且成纤维细胞增殖处于明显旺盛状态。结论促长散具有加速创面愈合的作用。     

康复新液促进小鼠压疮创面愈合机制的研究

目的 探讨康复新液对小鼠压疮创面促进愈合的作用及机制.方法 根据缺血-再灌注损伤机制,通过体外磁片加压法制备小鼠早期压疮模型,观察康复新液对小鼠压疮愈合的影响和组织病理学改变,通过免疫组织化学法从碱性成纤维细胞生长因子(bFGF)、转化生长因子(TGF-β)、表皮细胞生长因子(EGF)的蛋白表达方面研究康复新液的作用机理.结果 康复新液对小鼠压疮具有一定的促愈作用,可促进肉芽组织生长、血管新生、坏死组织、炎性物质的清除,较高剂量的康复新液可上调bFGF、TGF-β、EGF的蛋白表达,较低剂量的康复新液亦可促进bFGF的蛋白表达.结论 康复新液对模型小鼠早期压疮创面促愈和修复的作用机制可能与促进bFGF蛋白的表达相关,对TGF-β及EGF蛋白表达的上调可能也发挥了协同作用.     

无机活性元素对大鼠慢性溃疡组织乏氧诱导因子-1α及细胞因子的影响及机制

目的检测无机活性元素(德莫林)对大鼠皮肤慢性溃疡组织的影响,并对其作用机制进行分析。方法制备SD大鼠慢性溃疡模型,实验组应用无机活性元素,对照组常规消毒换药,另设空白组。观察3组溃疡的愈合时间。应用RT-PCR技术对3组溃疡组织乏氧诱导因子(HIF)-1α、表皮细胞生长因子(EGF)、碱性成纤维细胞因子(bFGF)、血管内皮细胞生长因子(VEGF)的表达进行检测,并对其差异进行比较分析。结果与对照组比较,实验组创面面积缩小更为明显(P<0.05);实验组、对照组的HIF-1α、EGF、bFGF和VEGF表达均明显高于正常皮肤(P<0.05),实验组EGF、bFGF和VEGF表达高于对照组,而HIF-1α表达低于对照组(P<0.05)。结论无机活性元素对慢性溃疡的治疗效果优于传统换药方法,其机制与该药物能促进EGF、bFGF和VEGF表达、抑制HIF-1α表达有关。     

Effect of inorganic active element on hypoxia inducing factor-1α and cytokines in chronic ulcer tissue in rats and its action mechanism

目的 检测无机活性元素(德莫林)对大鼠皮肤慢性溃疡组织的影响,并对其作用机制进行分析.方法 制备SD大鼠慢性溃疡模型,实验组应用无机活性元素,对照组常规消毒换药,另设空白组.观察3组溃疡的愈合时间.应用RT-PCR技术对3组溃疡组织乏氧诱导因子(HIF)-1α、表皮细胞生长因子(EGF)、碱性成纤维细胞因子(bFGF)、血管内皮细胞生长因子(VEGF)的表达进行检测,并对其差异进行比较分析.结果 与对照组比较,实验组创面面积缩小更为明显(P＜0.05);实验组、对照组的HIF-1α、EGF、bFGF和VEGF表达均明显高于正常皮肤(P＜0.05),实验组EGF、bFGF和VEGF表达高于对照组,而HIF-1α表达低于对照组(P＜0.05).结论 无机活性元素对慢性溃疡的治疗效果优于传统换药方法,其机制与该药物能促进EGF、bFGF和VEGF表达、抑制HIF-1α表达有关.     

芦荟凝胶对人皮肤成纤维细胞的增殖作用及其机制研究

目的研究芦荟凝胶对人皮肤成纤维细胞增殖的影响及其作用机制。方法体外培养人皮肤成纤维细胞,MTT法测定芦荟凝胶作用后人皮肤成纤维细胞的增殖能力,用实时荧光定量PCR法检测Ⅰ、Ⅲ型胶原mRNA水平,用免疫蛋白印迹法检测Ⅰ、Ⅲ型胶原蛋白的表达。结果 MTT检测结果表明,芦荟凝胶对人皮肤成纤维细胞具有促增殖作用,且呈现时间依赖性和剂量依赖性。芦荟凝胶作用于人皮肤成纤维细胞72h后,Ⅰ、Ⅲ型胶原mRNA及蛋白的表达显著增加(P<0.05,P<0.01)。结论芦荟凝胶可通过促进人皮肤成纤维细胞增殖以及Ⅰ、Ⅲ型胶原mRNA和蛋白的表达加速创面愈合。     

苦参素对成纤维细胞增殖、形态学及转化生长因子β_1的影响

目的 :研究苦参素对成纤维细胞增殖、形态学及转化生长因子β1(TGF β1)表达的影响 ,阐明其抗肝纤维化的作用机制。方法 :用甲基噻唑基四唑MTT(methylthiazolyltetrazolium)比色法、HE染色及免疫细胞化学技术分别检测小鼠皮肤成纤维细胞(NIH3T3)增殖、形态学及TGF β1的表达。结果 :苦参素能明显抑制成纤维细胞增殖及TGF β1的表达 ,并呈剂量依赖性。结论 :苦参素可通过抑制成纤维细胞增殖及TGF β1的表达而起到抗肝纤维化作用。     

短肽支架纤维对创面表皮碱性成纤维生长因子及表皮细胞生长因子的促进作用

目的：探讨自组装纤维支架肽RADA16在皮肤烫伤中的治疗作用。方法：在SD大鼠背部以电力机械方法制造深Ⅱ度烧伤模型,对照治疗组以0.9%NaCl溶液代替,烧伤后按时换药,并在不同时间段（伤后7、10、14d分别取烧伤修复创面的皮肤组织进行免疫组化染色,记录创面碱性成纤维细胞生长因子（bFGF）及表皮细胞生长因子（EGF）的表达,并以图像处理系统软件记录生长因子表达的半定量检测。比较了短肽处理组及空白对照组创面生长因子表达的灰度值。结果：免疫组织化学结果显示,在烧伤修复的不同修复期,短肽处理组bFGF和EGF均明显表达在新生表皮组织中,与空白对照组比较,差异均有统计学意义（P〈0.05）。结论：自组装纤维支架肽对烧伤皮肤生长因子的表达具有促进作用。     

肝素和碱性成纤维细胞生长因子对软骨细胞体外培养的作用研究

目的探讨肝素与碱性成纤维细胞生长因子(bFGF)对兔软骨细胞的增殖影响及其协同作用。方法体外培养3周龄的新西兰大白兔的原代关节软骨细胞。将第二代软骨细胞加入不同浓度的肝素、碱性成纤维细胞生长因子(bFGF)及肝素 bFGF联合应用,于48及72小时分别进行MTT检测软骨细胞的成活数。结果bFGF浓度在10ng／ml时即可显著促进软骨细胞增殖,当浓度增至50ng／ml,其促进作用达到最大值;单纯使用肝素并不促进软骨细胞增殖,当联用bFGF,bFGF浓度为50ng／ml、肝素浓度为1250ng／ml时协同作用最明显。结论bFGF可明显促进软骨细胞增殖,单用肝素对细胞增殖无明显影响,与bFGF联用时呈现协同效应,具有增强bFGF促软骨细胞增殖作用。     

半边旗二萜类化合物5F体外抗支气管哮喘小鼠气道纤维化的作用

目的观察半边旗二萜类化合物5F对体外培养的支气管哮喘小鼠气道成纤维细胞增殖和胶原合成及结缔组织生长因子(CTGF)表达的影响.方法将不同浓度(8,32,128 mg·L-1)的5F作用于体外培养的支气管哮喘小鼠气道成纤维细胞,通过噻唑蓝比色法(MTT)检测5F对细胞增殖的作用;逆转录聚合酶链反应(RT-PCR)检测其对细胞中CTGF mRNA表达水平的影响;化学比色法检测其对细胞胶原蛋白含量的影响.结果半边旗5F对支气管哮喘小鼠气道成纤维细胞的增殖具有显著的抑制作用,还能显著降低胶原蛋白的含量(P＜0.01);细胞中的CTGF mRNA的表达水平显著下调,并且均呈现量效的依赖关系.结论半边旗5F具有体外抗支气管哮喘小鼠气道纤维化的作用,这为寻找有效治疗哮喘的药物提供了新的思路.     

载碱性成纤维细胞生长因子质粒纳米粒的制备及体外转染活性研究

目的:制备壳聚糖载bFGF基因纳米粒,并对其体外性质及转染成纤维细胞效率进行考察。方法:以复凝集法制备载绿色荧光蛋白(EGFP)与人碱性成纤维细胞生长因子(bFGF)融合蛋白EGFP-bFGF质粒(pEGFP/bFGF)的壳聚糖纳米粒CS-pEGFP/bFGF,透射电镜、纳米粒度电位仪测定纳米粒形态、粒径和表面电位,凝胶阻滞实验分析质粒与壳聚糖结合情况,细胞增殖实验考察载bFGF质粒转染后对细胞增殖的影响,荧光分光光度计及荧光显微镜观测纳米粒转染成纤维细胞后细胞对EGFP的表达情况。结果:壳聚糖纳米粒可将质粒pEGFP/bFGF成功转入成纤维细胞中并能有效降低转染产生的细胞毒性,细胞自分泌的bFGF能促进成纤维细胞的增殖。结论:载bFGF纳米粒具有提高转染效率、降低毒性及促进成纤维细胞增殖的作用。     

BFGF、EGF联合应用对新生大鼠海马神干细胞分化的影响

目的:探讨EGF和bFGF联合应用后对新生大鼠海马神经干细胞增殖分化的影响。方法:联合应用EGF和bFGF培养海马神经干细胞及诱导贴壁的细胞分化,并用免疫细胞化学技术研究神经干细胞的分化情况。结果:bFGF和EGF联合应用可以明显扩增海马神经干细胞,并诱导细胞向神经元和星形胶质细胞分化。结论:bFGF和EGF除了具有促神经干细胞有丝分裂作用外,还可以诱导神经干细胞向神经元和星形胶质细胞分化。     

Thrombin-stimulated proliferation of cultured human synovial fibroblasts through proteolytic activation of proteinase-activated receptor-1

We examined the mechanism of thrombin on proliferation of synovial fibroblasts obtained from rheumatoid arthritis (RA). Thrombin concentration-dependently induced proliferation of synovial fibroblasts. Proliferation in response to thrombin (10 U/ml) was completely blocked by hirudin. TP367 and TP508, peptides corresponding to 2 noncatalytic regions of thrombin, failed to induce cell proliferation. Thrombin did not induce the production of basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF), and epidermal growth factor (EGF) in synovial fibroblasts. Expression of proteinase-activated receptor (PAR)-1 and PAR-3 mRNAs was observed in synovial fibroblasts. Thrombin and PAR-1 agonist peptide (AP), but not PAR-3 AP, induced intracellular calcium mobilization. PAR-1 AP induced cell proliferation whereas PAR-3 AP and PAR-4 AP had no effect on proliferation. Pertussis toxin (PTX), a Gialpha protein inhibitor; wortmannin, a PI (phosphatidylinositol) 3-kinase inhibitor; and PD98059, a specific MEK [mitogen-activated protein (MAK) kinase kinase] inhibitor, inhibited the thrombin-induced cell proliferation. Furthermore, the proliferation of synovial fibroblasts was suppressed by U-73122, a PLC (phospholipase C) inhibitor; 2-APB, an antagonist of InsP3 (inositol 1,4,5-triphosphate) receptor; and GF-109203X, a PKC (protein kinase C) inhibitor. These results suggest that thrombin induces the proliferation of RA synovial fibroblasts through the activation of PAR-1, leading to the PTX-sensitive G proteins - PI3 kinase pathway and PTX-insensitive G proteins - PLC (InsP3 receptor) Ca(2+)-PKC branch.     

Inhibition of human dermal fibroblast proliferation by removal of dermatan sulfate

In the current study, a glycosaminoglycan lyase, chondroitinase B, was used to study the role of dermatan sulfate proteoglycans on human dermal fibroblast proliferation. Pretreatment with chondroitinase B significantly decreased fibroblast proliferative responses to serum (20% to 55%). In contrast, heparinase III and chondroitinase AC were less effective in inhibiting fibroblast proliferation to serum. Analysis of glycosaminoglycans on chondroitinase B-treated fibroblasts confirmed that dermatan sulfate was removed from fibroblasts by this enzyme. Chondroitinase B treatment also decreased proliferation to basic fibroblast growth factor (bFGF) by 20% and reduced receptor binding by 25%. Heparinase III inhibited bFGF binding by 73%, but decreased proliferation to bFGF by only 21%. Chondroitinase AC had no effect on bFGF proliferation or binding. These data suggest that dermatan sulfate proteoglycans play a significant role in the control of human dermal fibroblast proliferation.     

EGF和bFGF对体外培养人视网膜色素上皮细胞增殖影响的比较

目的比较表皮生长因子（EGF）和碱性成纤维细胞生长因子（bFGF）对体外人视网膜色素上皮细胞（hRPE）增殖调控的剂量-效应关系、时间-效应关系,从而筛选出在体外培养hRPE增殖效应较强的生长因子,以提高体外hRPE的培养成效。方法采用经细胞化学染色法鉴定确认的hRPE细胞株,用CCK-8法检测成组配对的EGF和bFGF（0、2、4、8、16、32 ng/ml）在不同作用时间（0、24、48、72、96 h）对体外hRPE增殖的影响。结果在相同浓度EGF或bFGF作用下,hRPE吸光度值（A值）随作用时间延长而增加,与对照组比较的差异有统计学意义（P〈0.05）。在相同培养时间条件下,hRPE A值随EGF或bFGF浓度增加而增加,与对照组比较的差异有统计学意义（P〈0.05）;在相同作用时间下,浓度为4 ng/ml以上时,EGF组的hRPE吸光度值（A值）比bFGF组高且差异有统计学意义（P〈0.05）。结论 EGF和bFGF对体外培养hRPE增殖的调控有剂量-效应关系和时间-效应关系。同浓度EGF的促进细胞增殖效应强于bFGF。     

Inhibition by tranilast of nifedipine-induced proliferation of cultured human gingival fibroblasts

The appropriate method of etiologic therapy for gingival overgrowth is yet unknown. In this study drug-induced proliferation of Gin-1 cells, a normal human gingival fibroblast cell line, was examined by using the reagent water-soluble tetrazolium-1. Tranilast (100 microM) inhibited the nifedipine (10 microM)-induced proliferation of gingival fibroblasts. The level of basic fibroblast growth factor (bFGF) was determined by using an enzyme-linked immunosorbent assay kit. Tranilast inhibited the release of bFGF from the cells. In conclusion, tranilast depresses the nifedipine-induced proliferation of gingival fibroblasts by inhibiting the release of bFGF. Administration of tranilast may thus be clinically effective for the treatment of gingival overgrowth.     

Expression of Basic Fibroblast Growth Factor Correlates with Resistance to Paclitaxel in Human Patient Tumors

Background Preclinical results indicate acidic fibroblast growth factor (aFGF) and basic FGF (bFGF) present in solid tumors as a cause of broad-spectrum chemoresistance, whereas earlier clinical studies suggest that bFGF expression is associated with opposing outcomes in patients. We investigated the relationship between FGF expression and paclitaxel activity in tumors from bladder, breast, head and neck, ovarian, and prostate cancer patients. Materials and Methods Tumors (n = 96) were maintained in three-dimensional histocultures, retaining tumor–stromal interaction. Bladder tumors were treated with paclitaxel for 2 h, and the other tumors for 24 h. Antiproliferative and proapoptotic effects of paclitaxel were quantified and correlated with expression of aFGF, bFGF, P-glycoprotein (Pgp), p53, and bcl-2. Results Fifty-one percent (49/96) and 63% (61/96) of tumors showed aFGF and bFGF staining, respectively. aFGF expression was positively correlated with tumor stage (p < 0.01), and bFGF expression with tumor grade and Pgp expression (p < 0.05). Paclitaxel inhibited antiproliferation in 86% of tumors (83/96), with an average inhibition of 46 ± 19% (mean ± SD) in the responding tumors. Paclitaxel also induced apoptosis in 96% of tumors (92/96), with an average apoptotic index of 12 ± 7% in the responding tumors. aFGF expression did not correlate with tumor sensitivity to paclitaxel, whereas bFGF expression showed an inverse correlation (p < 0.01). bFGF expression was a stronger predictor of paclitaxel resistance compared to Pgp, p53, or Bcl-2. Conclusion These results support a role of bFGF in paclitaxel resistance in human patient tumors.     

bFGF和BDNF对MCAO大鼠海马区神经干细胞原位增殖的影响

目的：探讨碱性成纤维细胞生长因子（bFGF）、脑源性神经生长因子（BDNF）对大鼠脑缺血损伤后海马区（SGZ）神经干细胞原住增殖的影响。方法：将Wistar大鼠84只随机分为空白对照组（12只）、bFGF组（24只）、BDNF组（24只）、bFGF＋BD—NF组（24只）。采用线辁法制作局灶性大脑中动脉闭塞（MCAO）模型。分别于缺血再灌注后3d、7d、14d、28d处死大鼠。免疫组织化学方法动态检测SGZ区BrdU、Nestin的表达。结果：各组SGZ区的BrdU和Nestin阳性细胞均在脑缺血3d后开始增加,7d达到高峰,14d后开始下降,28d降至最低水平;与空白对照组相比,均有统计学意义（P〈0．05）;其中bFGF＋BDNF组Brdu和Nestin阳性细胞数增加更明显。结论：侧脑室注射bFGF、BDNF可促进脑缺血大鼠SGZ区内源性神经干细胞原位增殖;bFGF和EGF联合应用对脑缺血大鼠神经干细胞原位增殖效应有协同作用。     

Erythropoietin and growth factors exhibit differential angiogenic potential in mouse heart

BACKGROUND: The possible angiogenic effect of recombinant human erythropoietin (rHuEpo) and several possibly angiogenic cytokines such as basic fibroblast growth factor (bFGF), acidic fibroblast growth factor (aFGIF) and vascular endothelial growth factor (VEGF) was investigated in mouse heart. MATERIALS AND METHODS: Mice were divided into five groups (n = 7/group): A, control; B, rHuEpo-treated; C, (aFGF-treated); D, (VEGF-treated); E, (bFGF-treated). The antibody mouse anti-human CD31 was used to evaluate the vessels present in histological preparations. RESULTS: The results show a significant increase of the vessel number per optical field in the rHuEpo-treated, the bFGF-treated and the VEGF-treated animals compared to controls whereas aFGF did not show any significant angiogenic activity. CONCLUSION: Erythropoietin has a significant angiogenic effect in the mouse heart, similar to the effect of other angiogenic factors such as bFGF and VEGF whereas aFGF does not exhibit any effect.     

依济复与贝复济应用在供皮创面的比较

目的为了比较依济复与贝复济促进供皮区愈合的治疗效果。方法采用自身对照分组比较方法,观察供皮愈合时间,有无刺激性,所得数据用平均数(x±s)表示并χ2检验。结果对照组比较依济复及贝复济均能促进供皮区创面的愈合(P<0.01),依济复与贝复济比较其创面愈合时间无明显差异(P>0.05),两种药物共同使用在创面上与单独应用一种药物效果差异不大(P>0.05)。依济复与贝复济对创面都有轻度的刺激性。结论尽管作用对靶细胞有所不同,两种药物均能加速供皮创面的愈合,但两种药物对供皮区(取刃厚皮及中厚皮)的作用效果无明显差别。