成纤维细胞生长因子在辐射损伤防治中的研究进展

辐射损伤可由多因素引起,包括核辐射、放射性元素工业应用、电离辐射医学应用等,随着放射治疗、介入治疗等医学应用的迅速发展,辐射损伤的防治显得尤为重要。辐射可导致急性、慢性的组织损伤甚至严重的器官功能障碍,从而降低受照者生活质量甚至危及生命。成纤维细胞生长因子(FGF)作为一类有前景的辐射防护药物,为多种辐射损伤提供了新的防治手段,本文旨在针对具有辐射防治意义的几类FGF做一综述,主要包括FGF1、FGF2、FGF7等。     

肾细胞癌组织中b-FGF的表达意义和肿瘤微血管密度研究

探讨碱性成纤维细胞生长因子（b－FGF）在肾癌中表达的意义和其在肾癌微血管形成中的作用及与肾癌生物学特性的关系，采用斑点杂交技术测定20例肾癌患癌组织、癌旁和正常肾组织b－FGF表达情况；用免疫组化技术测定35例肾癌患者癌组织、癌旁和正常肾组织b－FGF表达和肿瘤微血管密度（MVD）。结果显示，肾癌组织中b－FGF表达明显高于癌旁和周围正常肾组织（P＜0．01）。b－FGF表达升高与肿瘤分明、分级密切相关；肾细胞癌组织中MVD与b－FGF表达呈明显正相关。提示b－FGF是肾细胞癌组织中重要的血管生长因子之一，b－FGF表达和MVD与肾细胞癌的预后有关。     

成纤维细胞生长因子7、10及其受体2Ⅲb在小鼠肾脏发育中的表达

目的观察成纤维细胞生长因子7、10(FGF7、FGF10)及其受体2Ⅲb(FGFR2Ⅲb)在不同发育阶段的小鼠肾脏的表达变化情况,探讨其与肾脏发育的关系。方法选取胚龄E12、14、16、18d胎鼠和生后N1、3、7、14、21和42d仔鼠肾脏,应用免疫组织化学技术对小鼠肾组织中FGF7、FGF10和FGFR2Ⅲb表达进行定位观察;应用体视学方法及Western blotting法对小鼠肾组织中FGF7、FGF10和FGFR2Ⅲb表达进行定性分析和定量检测。结果免疫组织化学法显示,E12d时,FGF7、FGF10和FGFR2Ⅲb开始在输尿管芽微弱表达;随着肾脏发育成熟,三者主要表达于远端小管和集合管。FGF10在近曲小管表达,近直小管无表达;FGF7和FGFR2Ⅲb在近端小管无表达。生后肾组织及发育各阶段肾小体未见FGF7、FGF10和FGFR2Ⅲb的表达。体视学和Western blotting结果显示,从E14d至N7d,三者在肾脏的表达量明显增加,N7d至N42d表达量逐渐减少。结论 FGF7、FGF10和FGFR2Ⅲb在肾脏发生和发育过程中可能起着重要作用。FGF10主要作用于近曲小管、远端小管和集合管,而FGF7和FGFR2Ⅲb主要作用于远端小管和集合管。     

HGF和FGF4在诱导骨髓间充质干细胞向肝细胞分化中的作用

目的 评估肝细胞生长因子(HGF)和成纤维细胞生长因子4(FGF4)在诱导骨髓间充质干细胞向肝细胞分化中的作用.方法 CCl_4经腹腔注射复制小鼠肝损伤模型,48h后处死小鼠取肝组织,分离肝脏组织制备肝损伤条件培养液进行培养.以正常小鼠肝组织制备的培养液作为对照组.ELISA法测定培养0、1、5、3、6、12、24、48h后肝组织培养液中的HGF和FGF4含量.用肝损伤条件培养液培养骨髓间充质干细胞,分别加入HGF抗体和(或)FGF4抗体,以封闭培养液中的HGF和(或)FGF4因子,未添加细胞因子抗体的培养液作为对照组;ELISA法检测肝细胞特异性白蛋白(ALB)水平,免疫荧光法检测细胞内ALB、甲胎蛋白(AFP)、细胞角蛋白19(CK19)的表达,评价骨髓间充质干细胞向肝细胞的分化状况.结果 与对照组比较,肝损伤小鼠肝细胞培养液中HGF和FGF4表达量均升高(P<0.05);在培养3h时HGF、FGF4上升达峰值,随后下降;FGF4在培养12h达低谷后再次升高.抑制HGF和(或)FGF4第14天后,ELISA和荧光标记染色法检测均显示ALB表达下降,与对照组比较差异显著(P<0.05);荧光标记染色显示封闭后AFP及CK-19表达均有下降(P<0.05).结论 HGF、FGF4是诱导骨髓间充质干细胞向肝细胞分化的重要因子,且可能有其他因子参与诱导肝细胞的定向分化.     

成纤维细胞生长因子促进骨再生的研究进展

成纤维细胞生长因子（fibroblast growth factors，FGF）是一组广泛存在于机体组织内具有相似结构特征的多肽，随着基因分离技术的发展与应用，目前已知FGF家族至少包括23个可编码相关结构蛋白的基因，因其具有很强的促细胞分裂．增殖活性以及能通过细胞因子网络发挥广泛的生物学作用．在骨科领域的应用已成为近年来的研究热点之一，现就FGF在促进骨再生方面的作用及应用报道如下。     

成纤维细胞生长因子生物蛋白海绵在创伤愈合中的应用

成纤维细胞生长因子（FGF）是一类体内含量较少但广泛存在的活性多肽类物质，能够有效地刺激和趋化成纤维细胞向胶原蛋白和肉芽组织发展，进而促进创伤的修复与愈合。而FGF总量相同的情况下，以持续低量用药的方式创伤组织的愈合优于早期或晚期大剂量用药的方式。由FGF和胶原蛋白复合而成的新型医用活性材料生物蛋白海绵（商品名：创必复），     

成纤维细胞生长因子10对人角朊细胞表达GM-CSF的刺激作用

研究FGF 10对角朊细胞表达GM CSF的作用 ,以探讨FGF 10促进创面愈合的机制。按FGF 10的浓度分为 4、16、12 5和5 0 0ng/ml四组 ,细胞接种密度为低密度 (2 5 0 0细胞 /cm2 )和高密度 (5 0 0 0细胞/cm2 )两种 ,分别于FGF 10作用后 2 4、48和 72h收集细胞培养液上清并进行细胞计数 ,测定GM CSF的含量。结果显示 ,低密度接种时 ,2 4h收集的各组上清中均未能检测到GM CSF,48h上清中的12 5ng/ml和 5 0 0ng/mlFGF 10组GM CSF的浓度及单个细胞的GM CSF的分泌均显著高于阴性对照组(P <0 0 5 )。 72h的培养上清中仅5 0 0ng/mlFGF 10组GM CSF的分泌量显著高于对照组 (P<0 0 5 )。高密度接种时 ,2 4h的上清中 16、12 5、5 0 0ng/ml的FGF 10各组GM CSF浓度显著高于对照组 (P<0 0 5 ) ,但单个细胞GM CSF的分泌无显著性变化 (P >0 0 5 ) ,48h的培养上清中 ,FGF 10各组的GM CSF均未升高 ,且 48h各组单个细胞GM CSF的分泌与细胞总数呈负相关 (r2 =0 881,P <0 0 5 )。结果提示FGF 10可能通过刺激GM CSF的分泌来促进创面肉芽组织形成。     

组织器官的原位再生复制研究报告

本研究报告,重点报道了组织器官的原位再生复制的临床程序,报道了组织潜能再生细胞的发现和存在,以及该细胞的增殖分化和形成组织器官的变化规律。以烧伤后皮肤组织器官的原位再生复制为模型,研究出了体外组织潜能再生细胞复制组织器官的培养方法。以体外组织器官的复制为模型．建立了寻找原位组织器官再生复制所需生命物质的方法和技术。本研究,首先按人体的器官功能分解为206个功能单位,确立了所复制的人体器官中的组织功能单位为组织器官,从而建立了原位组织器官再生复制的组织学基础。为了验证组织潜能再生细胞的再生潜能,建立了皮肤器官原位再生的实体临床跟踪技求,同时又建立了能代表有关器官功能类别的代表组织器官的原位和体外复制模型,以多组织器官的成功复制确定潜能再生细胞的作用．确定生命再生物质的重要性．确定组织器官原位再生复制的可行性,确定了组织器官原位再生复制的生命科学研究和医学进步的重大应用价值．同时展示了用此方法和技术攻克癌症的前景。本项研究报告,以近二百幅多个组织器官原位和体外再生复制的实体图片,展示了潜能再生细胞复制的组织器和和大器官实体;展示了细胞再生复制器官的全过程。真实的报告了组织器官原位再生复制的成果,所公布的主要成果为：皮肤器官的原位再生复制;胃肠黏膜组织器官的原位和体外再生复制;毛囊组织器官的原位和体外再生复制：神经组织器官的原位复制;胰腺组织器官的体外复制;骨髓组织的体外复制;肾小球肾小管组织器官的体外复制;心肌的体外复制等。为了让更多的人学会和掌握组织器官原位再生复制技术,木报告首次公布了实施技术的重要环节和技术流程;首次公布了生命再生物质的框架性结构和组成。作者自我认为,这是作者自费研究成果对人类生命科学的一大贡献。     

EPO促进血管痴呆大鼠神经干细胞增殖的机制研究

目的研究促红细胞生成素（EPO）促进血管性痴呆（VD）大鼠神经干细胞（NSC）增殖的分子调控机制。方法选取SD大鼠60只,采用改良血管阻断加硝普钠降压法建立VD大鼠模型,随机分为VD组和治疗组;治疗组腹腔注射EPO,分别于建模后7、14 d提取分离各组大鼠脑组织RNA,采用RT-PCR检测NSC增殖相关调控基因FGF2、EGF、TGFа表达。结果治疗组7 d、14 d时的FGF2、EGF表达均高于VD组（P〈0.05）,TGFа表达两组无统计学差异;7 d时两组FGF2表达明显高于EGF（P〈0.05）,14 d时则无统计学差异。结论 EPO通过上调胞外FGF2及EGF表达促进VD大鼠NSC增殖;在VD大鼠NSC增殖早期,FGF2起主要作用。     

中国人镧系元素膳食摄入量和主要器官及组织负荷量研究

目的 测定我国成年男子膳食食品和主要器官、组织中镧系元素浓度和估算其膳食日摄入量和器官、组织负荷量。方法 在我国不同膳食类型4地区采集21例急死正常尸体10种主要器官、组织样品，连同过去所采当地食品和31例尸体6种器官、组织样品，采用ICP—MS或INAA法及必要的质量控制措施，分别测定了11和14种镧系元素浓度。按当地膳食组成和中同参考人器官组织重量估算了摄入量和器官、组织负荷量。结果 获得了我国12类食品和10种主要器官、组织样品中14种镧系元素浓度、摄入量和相应器官、组织负荷量。结论 本研究首次获得我国各类食品和成年男子10种器官组织中11种镧系元素浓度、相应膳食口摄入量和负荷量。所获结果为确定中国参考人镧系元素相应参数提供较前更为系统的依据，也为不同镧系元素、食品和器官组织提供了比较和背景值的国情资料。     

碱性成纤维细胞生长因子对心肌细胞间信号转导的影响

目的 观察碱性成纤维细胞生长因子(FGF-2)对心肌细胞间信号转导的作用。方法 采用激光黏附细胞扫描仪和荧光光淬灭后恢复技术(FRAP),观察FGF-2作用于心肌细胞1和4天后胞间信号转导的变化。结果 10～100ng／ml的FGF-2能够抑制原代培养的心肌细胞的胞间信号转导,同时促进细胞增生;高浓度的FGF-2(1000ng／ml)使心肌细胞的胞间信号转导明显增加,但抑制细胞生长。结论 FGF-2可以影响心肌细胞的胞间信号转导,可能与其参与心肌细胞的生长、发育甚或凋亡等生物过程有关。     

FGF2 mediates DNA repair in epidermoid carcinoma cells exposed to ionizing radiation

Abstract Purpose: Fibroblast growth factor 2 (FGF2) is a well-known survival factor. However, its role in DNA repair is poorly documented. The present study was designed to investigate in epidermoid carcinoma cells the potential role of FGF2 in DNA repair. Materials and methods: The side population (SP) with cancer stem cell-like properties and the main population (MP) were isolated from human A431 squamous carcinoma cells. Radiation-induced DNA damage and repair were assessed using the alkaline comet assay. FGF2 expression was quantified by enzyme linked immunosorbent assay (ELISA). Results: SP cells exhibited rapid repair of radiation induced DNA damage and a high constitutive level of nuclear FGF2. Blocking FGF2 signaling abrogated the rapid DNA repair. In contrast, in MP cells, a slower repair of damage was associated with low basal expression of FGF2. Moreover, the addition of exogenous FGF2 accelerated DNA repair in MP cells. When irradiated, SP cells secreted FGF2, whereas MP cells did not. Conclusions: FGF2 was found to mediate DNA repair in epidermoid carcinoma cells. We postulate that carcinoma stem cells would be intrinsically primed to rapidly repair DNA damage by a high constitutive level of nuclear FGF2. In contrast, the main population with a low FGF2 content exhibits a lower repair rate which can be increased by exogenous FGF2.     

FGF2 mediates DNA repair in epidermoid carcinoma cells exposed to ionizing radiation

Abstract

Purpose: Fibroblast growth factor 2 (FGF2) is a well-known survival factor. However, its role in DNA repair is poorly documented. The present study was designed to investigate in epidermoid carcinoma cells the potential role of FGF2 in DNA repair.

Materials and methods: The side population (SP) with cancer stem cell-like properties and the main population (MP) were isolated from human A431 squamous carcinoma cells. Radiation-induced DNA damage and repair were assessed using the alkaline comet assay. FGF2 expression was quantified by enzyme linked immunosorbent assay (ELISA).

Results: SP cells exhibited rapid repair of radiation induced DNA damage and a high constitutive level of nuclear FGF2. Blocking FGF2 signaling abrogated the rapid DNA repair. In contrast, in MP cells, a slower repair of damage was associated with low basal expression of FGF2. Moreover, the addition of exogenous FGF2 accelerated DNA repair in MP cells. When irradiated, SP cells secreted FGF2, whereas MP cells did not.

Conclusions: FGF2 was found to mediate DNA repair in epidermoid carcinoma cells. We postulate that carcinoma stem cells would be intrinsically primed to rapidly repair DNA damage by a high constitutive level of nuclear FGF2. In contrast, the main population with a low FGF2 content exhibits a lower repair rate which can be increased by exogenous FGF2.     

Angiogenic effect of intramuscular administration of basic and acidic fibroblast growth factor on skeletal muscles and influence of exercise on muscle angiogenesis

Background

Angiogenic factors which control the angiogenic process represent a promising strategy for restoration of blood flow, but require further evaluation before clinical use. Exercise has also been reported to induce neovascularisation in muscles.

Objectives

To evaluate the angiogenic effects of basic fibroblast growth factor (b‐FGF) and acidic fibroblast growth factor (a‐FGF) on rat gastrocnemius muscle, when administered intramuscularly, and to compare them with those obtained by daily exercise.

Methods

Forty nine rats were allotted to the following groups: A, controls; B, exercise by swimming; C1 and C2, intramuscular injection of b‐FGF and a‐FGF respectively; D1 and D2, b‐FGF and a‐FGF injection in combination with exercise. The antibody mouse anti‐rat CD31 was used to evaluate the numbers of blood vessels present in histological preparations of gastrocnemius muscle.

Results

Significant increases in the numbers of blood vessels of the right gastrocnemius muscles in groups C1 and D1 were observed compared with controls (p<0.05). There was only a slight increase in angiogenesis in the left gastrocnemius muscle of groups C1 and D1 compared with controls (p>0.05), and there was a decrease in angiogenesis in the gastrocnemius muscle of the swimming group compared with controls.

Conclusion

The intramuscular administration of b‐FGF, but not a‐FGF, induced significant local angiogenesis in gastrocnemius muscle at the site of injection.     

碱性成纤维细胞生长因子与纤维蛋白黏合剂对半月板无血运区损伤的修复作用

目的 研究碱性成纤维细胞生长因子（ｂＦＧＦ）与纤维蛋白黏合剂（ＦＴＡ）对半月板无血运区损伤的修复作用。方法 选用健康成年青紫兰兔６只,在兔半月板上造成统一的无血运区损伤,随机将兔分成３组,分别为空白且、ＦＴＡ治疗组和ＦＧＦ／ＦＴＡ治疗组。术后２,６,１２周分批处死动物,进行大体形态观察和组织学检查。结果 ＦＴＡ治疗组能形成癜痕组织愈合;ＦＧＦ／ＦＴＡ治疗组为纤维软骨样组织愈合,但其愈合组织与正常半     

Angiogenic effect of intramuscular administration of basic fibroblast growth factor in atrophied muscles: an experimental study in the rat

Background

Although angiogenic therapy using recombinant growth factors holds much hope for the treatment of ischaemic diseases, there are still many unanswered questions, including its effectiveness on atrophic muscles.

Objective

To evaluate the angiogenic effects of intramuscularly administered basic fibroblast growth factor (b‐FGF) on normal gastrocnemius muscles of rats and atrophic gastrocnemius muscles after tenotomy.

Methods

Forty rats were divided into groups as follows: group A, controls; group B, injected with 1 μg b‐FGF; group C, tenotomy performed on the right gastrocnemius muscle; group D, tenotomy and 1 μg b‐FGF. Mouse anti‐rat CD31 antibody was used to evaluate the number of blood vessels present in histological preparations.

Results

There was a significant (p<0.01) decrease in the number of blood vessels compared with the controls in the atrophic muscles of group C. This was similar to the decrease in muscle weight in this group. However, there was a significant (p<0.01) increase in the number of blood vessels compared with the controls in groups B and D. Similarly, there was a significant (p<0.01) increase in the number of blood vessels in group D compared with the atrophic muscles in group C.

Conclusion

Intramuscular administration of b‐FGF increases angiogenesis in both normal and atrophic rat gastrocnemius muscles at the injection area.     

Effect of keratinocyte growth factor on the proliferation,clonogenic capacity and colony size of human epithelial tumour cells in vitro

Purpose : The effect of recombinant human keratinocyte growth factor (rHuKGF) on the proliferation, clonogenic capacity and colony size of low-passage human epithelial tumour cells was tested in vitro. Materials and methods : Five tumour cell cultures derived from head and neck squamous cell carcinomas, three cultures derived from pleural effusions of carcinomas of different origin and normal human nasal epithelial cells were analysed in passages 2-4. Expression of FGF7 and its receptor (FGFR2) were determined by the RNase protection assay. Cells were incubated with rHuKGF (10-200 ng ml ?1) 3 days before or immediately after plating for clonal growth in serum-depleted media. To determine cellular radiosensitivity, single doses of 1-8 Gy X-rays were applied. Colony formation as well as colony size, reflecting the number of cell divisions, was determined after 10-15 days of growth in rHuKGF-treated and control cells. Results : Normal nasal epithelial cells showed a two- to threefold increase in the number of cell divisions due to rHuKGF-treatment. In tumour cell cultures, significant stimulation of proliferation occurred in only one of eight samples. Tumour cells expressed FGF7 mRNA and protein, and low levels of FGFR2 mRNA. The addition of rHuKGF to the medium of the tumour cell cultures influenced neither radiation-induced impairment of proliferation nor clonogenic cell survival. Conclusion : rHuKGF has been shown to ameliorate the radiation tolerance of normal epithelia. The minimum in vitro tumour cell response to rHuKGF compared with normal epithelial cells suggests a potential for selective protection of normal epithelia during radiotherapy. The low FGFR2 expression as well as the FGF7 expression in the tumour cells may contribute to their resistance to rHuKGF treatment.     

Effect of keratinocyte growth factor on the proliferation,clonogenic capacity and colony size of human epithelial tumour cells in vitro

PURPOSE: The effect of recombinant human keratinocyte growth factor (rHuKGF) on the proliferation, clonogenic capacity and colony size of low-passage human epithelial tumour cells was tested in vitro. MATERIALS AND METHODS: Five tumour cell cultures derived from head and neck squamous cell carcinomas, three cultures derived from pleural effusions of carcinomas of different origin and normal human nasal epithelial cells were analysed in passages 2-4. Expression of FGF7 and its receptor (FGFR2) were determined by the RNase protection assay. Cells were incubated with rHuKGF (10-200 ng ml(-1)) 3 days before or immediately after plating for clonal growth in serum-depleted media. To determine cellular radiosensitivity, single doses of 1-8 Gy X-rays were applied. Colony formation as well as colony size, reflecting the number of cell divisions, was determined after 10-15 days of growth in rHuKGF-treated and control cells. RESULTS: Normal nasal epithelial cells showed a two- to threefold increase in the number of cell divisions due to rHuKGF-treatment. In tumour cell cultures, significant stimulation of proliferation occurred in only one of eight samples. Tumour cells expressed FGF7 mRNA and protein, and low levels of FGFR2 mRNA. The addition of rHuKGF to the medium of the tumour cell cultures influenced neither radiation-induced impairment of proliferation nor clonogenic cell survival. CONCLUSION: rHuKGF has been shown to ameliorate the radiation tolerance of normal epithelia. The minimum in vitro tumour cell response to rHuKGF compared with normal epithelial cells suggests a potential for selective protection of normal epithelia during radiotherapy. The low FGFR2 expression as well as the FGF7 expression in the tumour cells may contribute to their resistance to rHuKGF treatment.