Effect of charge at an amino acid of basic fibroblast growth factor on its mitogenic activity

The amino acid at the 119th position of human basic fibroblast growth factor(hbFGF),lysine(K119),is a critical component for its mitogenic activity.However,little is known about the effects of the characteristics of this residue including charge on the mitogenic activity of hbFGF.Herein,this basic residue was replaced with neutral glutamine residue and acidic glutamic acid residue to construct mutants hbFGF^(K119Q) and hbFGF^(K119E),respectively.The mutants were produced by BL21(DE3)/pET3c expression system...

Basic fibroblast growth factor increases the numbe of endogenous neural stem cells and inhibits the expression of amino methyl isoxazole propionic acid receptors in amyotrophic lateral sclerosis mice

This study aimed to investigate the number of amino methyl isoxazole propionic acid （AMPA） receptors and production of endogenous neural stem cells in the SOD1 G93AG1H transgenic mouse model of amyotrophic lateral sclerosis, at postnatal day 60 following administration of basic fibroblast growth factor （FGF-2）. A radioligand binding assay and immunohistochemistry were used to estimate the number of AMPA receptors and endogenous neural stem cells respectively. Results showed that the number of AMPA receptors and endogenous neural stem cells in the brain stem and sensorimotor cortex were significantly increased, while motor function was significantly decreased at postnatal days 90 and 120. After administration of FGF-2 into mice, numbers of endogenous neural stem cells increased, while expression of AMPA receptors decreased, whilst motor functions were recovered. At postnatal day 120, the number of AMPA receptors was negatively correlated with the number of endogenous neural stem cells in model mice and FGF-2-treated mice. Our experimental findings indicate that FGF-2 can inhibit AMPA receptors and increase the number of endogenous neural stem cells, thus repairing neural injury in amyotrophic lateral sclerosis mice.

Exogenous acid fibroblast growth factor inhibits ischemia-reperfusion-induced damage in intestinal epithelium via regulating P53 and P21WAF-1 expression

AIM： To detect the effect of acid fibroblast growth factor （aFGF） on P53 and P21WAF-1 expression in rat intestine after ischemia-reperfusion （I-R） injury in order to explore the protective mechanisms of aFGF. METHODS： Hale rats were randomly divided into four groups, namely intestinal ischemia-reperfusion group （R）, aFGF treatment group （A）, intestinal ischemia group （I）, and sham-operated control group （C）. In group I, the animals were killed after 45 min of superior mesenteric artery （SHA） occlusion. In groups R and A, the rats sustained for 45 min of SHA occlusion and were treated with normal saline （0.15 mL） and aFGF （20 μg/kg, 0.15 mL）, then sustained at various times for up to 48 h after reperfusion. In group C, SHA was separated, but without occlusion. Apoptosis in intestinal villi was determined with terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling technique （TUNEL）. Intestinal tissue samples were taken not only for RT- PCR to detect P53 and P21WAF-1 gene expression, but also for immunohistochemical analysis to detect P53 and P21WAF-1 protein expression and distribution. RESULTS： In histopathological study, ameliorated intestinal structures were observed at 2, 6, and 12 h after reperfusion in A group compared to R group. The apoptotic rates were （41.17±3.49）%, （42.83±5.23）%, and （53.33±6.92）% at 2, 6, and 12 h after reperfusion, respectively in A group, which were apparently lower than those in R group at their matched time points （50.67±6.95）%, （54.17±7.86）%, and （64.33±6.47）%, respectively, （P〈0.05））. The protein contents of P53 and P21WAF-1 were both significantly decreased in A group compared to R group （P〈0.05） at 2-12 h after reperfusion, while the mRNA levels of P53 and P21VVAF-1 in A group were obviously lower than those in R group at 6-12 h after reperfusion （P〈0.05）. CONCLUSION： P53 and P21WAF-1 protein accumulations are associated with intestinal barrier injury induced by I-R insult, while intravenous aFGF can alleviate apoptosis of rat intestinal cells by inhibiting P53 and P21WAF-1 protein expression.

Fibroblast growth factor 15,induced by elevated bile acids,mediates the improvement of hepatic glucose metabolism after sleeve gastrectomy

BACKGROUND Fibroblast growth factor(FGF)15/19,which is expressed in and secreted from the distal ileum,can regulate hepatic glucose metabolism in an endocrine manner.The levels of both bile acids(BAs)and FGF15/19 are elevated after bariatric surgery.However,it is unclear whether the increase in FGF15/19 is induced by BAs.Moreover,it remains to be understood whether FGF15/19 elevations contribute to improvements in hepatic glucose metabolism after bariatric surgery.AIM To investigate the mechanism of improvement of hepatic glucose metabolism by elevated BAs after sleeve gastrectomy(SG).METHODS By calculating and comparing the changes of body weight after SG with SHAM group,we examined the weight-loss effect of SG.The oral glucose tolerance test(OGTT)test and area under the curve of OGTT curves were used to assess the anti-diabetic effects of SG.By detecting the glycogen content,expression and activity of glycogen synthase as well as the glucose-6-phosphatase(G6Pase)and phosphoenolpyruvate carboxykinase(Pepck),we evaluated the hepatic glycogen content and gluconeogenesis activity.We examined the levels of total BA(TBA)together with the farnesoid X receptor(FXR)-agonistic BA subspecies in systemic serum and portal vein at week 12 post-surgery.Then the histological expression of ileal FXR and FGF15 and hepatic FGF receptor 4(FGFR4)with its corresponding signal pathways involved in glucose metabolism were detected.RESULTS After surgery,food intake and body weight gain of SG group was decreased compare with the SHAM group.The hepatic glycogen content and glycogen synthase activity was significantly stimulated after SG,while the expression of the key enzyme for hepatic gluconeogenesis:G6Pase and Pepck,were depressed.TBA levels in serum and portal vein were both elevated after SG,the FXR-agonistic BA subspecies:Chenodeoxycholic acid(CDCA),lithocholic acid(LCA)in serum and CDCA,DCA,LCA in portal vein were all higher in SG group than that in SHAM group.Consequently,the ileal expression of FXR and FGF15 were also advanced in SG group.Moreover,the hepatic expression of FGFR4 was stimulated in SG-operated rats.As a result,the activity of its corresponding pathway for glycogen synthesis:FGFR4-Ras-extracellular signal regulated kinase pathway was stimulated,while the corresponding pathway for hepatic gluconeogenesis:FGFR4-cAMP regulatory element-binding protein-peroxisome proliferator-activated receptorγcoactivator-1αpathway was suppressed.CONCLUSION Elevated BAs after SG induced FGF15 expression in distal ileum by activating their receptor FXR.Furthermore,the promoted FGF15 partly mediated the improving effects on hepatic glucose metabolism of SG.

Role of Fibroblast Growth Factor 19 in Maintaining Nutrient Homeostasis and Disease

Fibroblast growth factor 19 （FGF19） is a kind of gut-derived postprandial hormone. As an atypical member of the FGF family, FGF19 functions as an endocrine hormone except regulating cell growth and differentiation. FGF19 plays a key role in coordination of liver bile acid biosynthesis and gallbladder motility, and acts as a regulator of metabolic homeostasis, including strengthening insulin sensitivity, decreasing triglyceride concentration and reducing body weight.

The change of basic fibroblast growth factor and effect of prostacyclin in lung injury

In this study,the changes in activity and mRNA transcript of basic fibroblast growth fac-tor(bFGF)in lung injury and the effect of prostacyclin on the treatment of lung injury wereinvestigated.The result of the experiment revealed that there were not any bioactive bFGF andbFGF mRNA in the lung tissue of normal dogs.bFGF activity and bFGF mRNA were detectedonly in the injured lung tissue.Prostacyclin could slightly elevate the activity of bFGF andsignificantly elevate the level of bFGF mRNA.The findings suggest that(1)the level of bFGF in-creased after lung injury.(2)Prostacyclin could influence the expression of bFGF.(3)bFGF couldplay an important role in the repair of injury lung tissue.

光子治疗仪联合rh-aFGF治疗小面积深Ⅱ度烧伤创面疗效观察

目的:探讨光子治疗仪联合重组人酸性成纤维细胞生长因子(Recombinant human acidic fibroblast growth factor,rh-aFGF)对小面积深Ⅱ度烧伤患者创面愈合、疼痛程度的影响。方法:选取2020年6月-2022年11月笔者医院收治的80例小面积深Ⅱ度烧伤患者,采用随机数字表法分为rh-aFGF组(n=40)和联合组(n=40)。rh-aFGF组给予rh-aFGF治疗,联合组给予光子治疗仪联合rh-aFGF治疗,比较两组治疗后10、15、19 d时的创面愈合率、创面肉芽生长评分及完全愈合时间;比较两组治疗后1、7、14 d时的创面疼痛数字量表(Numerical rating scale,NRS)评分;比较两组治疗后6个月时的温哥华瘢痕量表(Vancouver scar scale,VSS)评分。结果:联合组治疗后10、15、19 d创面愈合率、创面肉芽生长评分均高于rh-aFGF组,创面完全愈合时间短于rh-aFGF组(P<0.05)。两组治疗后1、7、14 d创面NRS评分均较治疗前降低,且联合组低于rh-aFGF组(P<0.05)。联合组治疗后6个月VSS评分低于rh-aFGF组(P<0.05)。结论:光子治疗仪联合rh-aFGF治疗小面积深Ⅱ度烧伤,可促进肉芽组织生长,加速创面愈合,减轻疼痛,减少瘢痕形成。

rh-aFGF凝胶联合点阵CO_(2)激光治疗对剖宫产术后切口瘢痕愈合和美观满意度的影响

目的:探讨重组人酸性成纤维细胞生长因子(Recombinant Human acidic fibroblast growth factor,rh-aFGF)凝胶联合点阵CO_(2)激光治疗对剖宫产术后切口瘢痕愈合和美观满意度的影响。方法:选取2020年6月-2022年6月在笔者医院收治的85例剖宫产术后皮肤瘢痕患者,根据治疗方法将其分为对照组(42例)和联合组(43例)。对照组在患者剖宫产术后3个月左右瘢痕出现后采取点阵CO_(2)激光治疗,联合组在对照组基础上予以rh-aFGF凝胶联合治疗。观察比较两组患者治疗前、治疗后1个月、治疗后3个月和治疗后6个月时的温哥华瘢痕量表(Vancouver scar scale,VSS)评分情况,治疗3个月时的临床治愈情况、美观满意度和不良反应发生情况。结果:治疗后1个月、3个月和6个月时,联合组VSS评分均低于对照组,且联合组临床总治愈率和美观满意度均较对照组高(P<0.05);联合组不良反应发生率低于对照组(P<0.05)。结论:相比于单纯使用点阵CO_(2)治疗,rh-aFGF凝胶联合点阵激光治疗对剖宫产术后切口瘢痕愈合效果更好,能有效提高患者对瘢痕恢复的满意程度,且不增加患者不良反应。

ACIDIC FIBROBLAST GROWTH FACTOR REDUCES RAT SKELETAL MUSCLE DAMAGE CAUSED BY ISCHEMIA AND REPERFUSION

Acute interruption of arterial blood flow to the extremities is often associated with significant morbidity and mortality. Broad spectrum mitogenic and non mitogenic activities of FGFs inspired us to study its protecting effects on tissue injuries in ischemia reperfusion condition. We found that systemic administration of aFGF after reperfusion onset prevented severe skeletal muscle injuries. In rats treated with aKGF, the tissue edema was reduced significantly, the tissue viability was increased, and the muscle fibers contained more succinate dehydrogenase (SDH) and adenosine triphosphatasc (ATPase). The pathological results supported the concept of improved prevention with aFGF treatment. The possible tissue protection by aFGF may come from its ability to regulate the concentration of evtra- and intracellular calcium ion. Besides, it may moderate other Ca2+ dependent enzyme conversion processes. Also, it may take part in the vascular tone regulation under ischemia and reperfusion conditions. These results suggest further study of tissue ischemia prevention with FGF and its possible mechanisms in the future.

Effect of basic fibroblast growth factor on the expression of glial fibrillary acidic protein after tractive spinal cord injury in rats

OBJECTIVE: To investigate the effects of basic fibroblast growth factor (bFGF) on the expression of glial fibrillary acidic protein (GFAP) after tractive spinal cord injury in rats and to explore the recovery of spinal cord function. METHODS: The rats were subjected to tractive spinal cord injury at T13-L2. Cortical somatosensory-evoked potential (CSEP) was closely monitored and when P1-N1 wave amplitude decreased to 70% of that before operation, a small-bore catheter was inserted below the injured plane through subarachnoid cavity. In the treatment groups, 20 microl of bFGF solution (containing 20 microg of bFGF) was injected through the catheter right after the operation and 1, 2, 3, 4, 8, 12 and 24 h postoperatively. In the control group, same volume of normal saline was injected and every four rats were killed at 1, 4, 7, 14 and 21 d after the operation. Combined behavior score (CBS) and electro-physiological examination were adopted to evaluate function recovery. Expression of GFAP was observed by immuno-histochemical staining and was analyzed quantitatively by computer image analysis. RESULTS: There was statistically significant difference in GFAP-positive cells between bFGF treatment group and the control group (P

aFGF植物表达载体构建及转化紫花苜蓿的初步研究

主要将aFGF基因按照植物偏爱的密码子进行基因优化,同时加上信号肽、6-组氨酸标签及凝血酶切割因子,合成全基因,再将基因插入到改造的TΩ4AB质粒中,将aFGF基因和质粒上的增强子、启动子、Ω序列及ployA加尾序列双酶切,命名为TΩ4AB-aF,将TΩ4AB-aF插入到pBI121载体中命名为pBI121-TΩ4AB-aF。利用三亲融合法将pBI121-TΩ4AB-aF转入到农杆菌菌株LBA4404中,转化苜蓿。转基因苗用卡那霉素进行筛选;并对抗性苗进行PCR、RT-PCR、Western Blot检测。结果表明,aFGF在苜蓿中得到表达。为苜蓿作为植物生物反应器奠定理论基础。

Genistein对aFGF诱导的AGZY-83A细胞增殖的抑制作用

目的 :探讨酪氨酸蛋白激酶 (TPK)抑制剂 Genistein对酸性成纤维细胞生长因子 (a FGF)诱导的肺腺癌细胞株 AGZY- 83A细胞增殖的抑制作用。方法 :以不同浓度的 a FGF刺激 AGZY- 83A细胞 ,再对由 a FGF引起增殖的细胞施加不同浓度的 Genistein,用细胞计数器计数细胞 ,观察 Genistein对细胞增殖的抑制作用。结果 :a FGF可使该细胞株增殖比明显增加 ,而 Genistein可引起细胞增殖比明显下降 ,其程度均随浓度增高而增强。结论 :该细胞株中 a FGF受体有 TPK活性 ,Genistein对 a

aFGF对大鼠胃粘膜溃疡的促愈合作用

目的 :观察酸性成纤维生细胞长因子 (aFGF)对大鼠胃粘膜溃疡愈合的影响。方法 :用醋酸酐诱发大鼠胃粘膜溃疡建立动物溃疡模型 ,以aFGF多克隆抗体作为抗体通过免疫组化方法观察aFGF在溃疡区的表达情况。结果 :aFGF在溃疡区肉芽组织细胞浆中有高度表达 ,aFGF使溃疡区中微血管数目明显增加。结论 :aFGF通过促进血管生成 ,及内皮细胞 成纤维细胞、平滑肌细胞。

抗菌肽AD与haFGF融合基因的合成及其表达

通过PCR技术和体外DNA重组技术将CecropinAD连接在haFGF改构体的 5′端 ,构建成融合基因CADAF。将其克隆到表达载体pET3c上 ,然后转化到BL2 1 (DE3)中经IPTG诱导表达。经DNA序列分析 ,合成的CADAF基因序列与设计序列一致。构建的CADAF基因在BL2 1 (DE3)

超声微泡靶向递送aFGF对糖尿病心肌病的治疗作用及其机制研究

目的:观察SonoVue超声微泡靶向递送酸性成纤维细胞生长因子(aFGF)对糖尿病心肌病(DCM)大鼠左室舒缩功能的保护作用并初步探讨其机制。方法:24只健康雄性SD大鼠通过腹腔注射链脲佐菌素建立DCM模型,再随机平均分成DCM组与aFGF治疗组。另选择正常对照组12只。aFGF治疗组经尾静脉注射SonoVue-aFGF溶液并同时给予心肌定点超声辐照。干预后4周对所有大鼠行心导管检查,测定左室收缩末压力(LVESP)、左室舒张末压力(LVEDP)和左室内压最大上升/下降速率(LV±dp/dt max)。处死大鼠取心肌组织,免疫组织化学染色检测心肌微血管密度(MVD),改良Masson胶原染色法测定心肌胶原容积分数(CVF),TUNEL法检测心肌组织凋亡指数(AI)。结果:干预后4周,aFGF治疗组大鼠LVESP和LV±dp/dt max与DCM组比较明显增加(P<0.01),LVEDP较DCM组明显减低(P<0.01)。aFGF治疗组MVD测值与DCM组比较明显增加(P<0.01),而CVF及AI较DCM组明显减低(P<0.01)。结论:超声微泡靶向递送aFGF可有效改善DCM大鼠的左心室功能,有望成为治疗DCM的新方法。

掌侧经皮Herbert螺钉内固定并注入aFGF治疗26例不稳定型腕舟骨骨折

目的:评价掌侧经皮Herbert螺钉内固定治疗不稳定型腕舟骨骨折的临床价值及酸性成纤维细胞生长因子(aFGF)对腕舟骨骨折愈合的作用。方法:26例不稳定型腕舟骨骨折患者,采用掌侧经皮Herbert螺钉内固定治疗,术中骨折处注入aFGF透明质酸凝胶。结果:26例中25例获随访,随访时间平均11 4个月。25例骨折全部愈合,愈合时间平均为术后3 6个月。25例伤腕没有疼痛,握力良好。2例腕关节活动轻度受限,另23例患侧腕关节桡偏、尺偏、背伸、掌屈活动与健侧相比无明显差别。结论:掌侧经皮Herbert螺钉内固定治疗不稳定型腕舟骨骨折具有创伤小、骨折固定牢靠、术后可早期功能锻炼、手腕功能恢复好、内固定无需取出等优点。术中加用aFGF透明质酸凝胶对骨折愈合可能有促进作用。

非促分裂haFGF对MNU所致大鼠视网膜损伤的保护作用

目的 研究非促分裂haFGF(nm haFGF)对N 甲基 N 亚硝脲 (MNU)所致大鼠视网膜损伤的保护作用及其作用机制。方法 通过ipMNU(60mg·kg-1 )复制大鼠视网膜损伤模型, 0和 12h后,玻璃体腔内注射不同剂量nm haFGF。24h和 7d后,测量周边视网膜总厚度和外视网膜厚度、光感受器细胞凋亡指数及视网膜Bcl 2和Bax蛋白表达水平。结果 MNU作用 24h后,nm haFGF组周边视网膜的凋亡指数显著降低 (P<0 01); 7d后,nm haFGF组周边视网膜光感受器细胞数明显增多,且周边视网膜总厚度和外视网膜厚度增加 (P<0 01);不同剂量nm haFGF可调节Bcl 2和Bax蛋白表达水平。结论 nm haFGF能部分抑制MNU引起的SD大鼠视网膜损伤,其作用机制可能与上调Bcl 2和下调Bax蛋白表达水平有关。

改构体aFGF对颈动脉窦损伤的保护作用

目的 探讨损伤大白鼠颈动脉窦压力感受器神经末梢后 ,改构体aFGF的保护作用。方法 将Wistar大鼠随机分成对照组、改构体aFGF1组、改构体aFGF2 组、改构体aFGF3 每组各 10只。改构体组分别经静脉注射 0 14 μg/ml、0 4 3μg/ml、1 30 μg/ml的改构体aFGF(1ml/ 10 0g) ,而对照组注射同剂量的生理盐水。 2 0min后 ,用蘸有 80 %乙醇的棉球轻轻擦拭右颈动脉窦区 ,损伤颈动脉窦压力感受器神经末梢 ,并放置 5min。观察记录各组损伤前后夹闭颈总动脉血压即 :收缩压 (SP)、舒张压(DP)、平均动脉压 (MP)和心率 (HR)的变化。结果 (1)损伤右颈动脉窦前 ,夹闭右颈总动脉 ,实验组和对照组血压均升高 ,夹闭前后相比差异均有统计学意义 (P <0 0 1) ;(2 )损伤右颈动脉窦后 ,夹闭右颈总动脉 ,改构体aFGF1组、对照组血压不变 ,夹闭前后相比差异无统计学意义 (P >0 0 5 ) ;改构体aFGF2 组、改构体aFGF3 组血压升高 ,夹闭前后相比差异有统计学意义 (P <0 0 5 ) ,夹闭前后两组的血压差分别与对照组的差值相比有统计学意义 (P <0 0 5 )。在本实验中 ,HR均无明显变化。结论 改构体aFGF对颈动脉窦损伤有保护作用 。

nmhaFGF对SD大鼠缺血再灌注视网膜SOD、MDA、NO的影响

目的观察非促分裂人酸性成纤维细胞生长因子(nmhaFGF)对大鼠视网膜缺血再灌注损伤后视网膜超氧化物歧化酶(SOD)、丙二醛(MDA)、一氧化氮(NO)的影响。方法40只SD大鼠随机分为对照组10只,实验组(nmhaFGF组)30只。实验组(大鼠左眼)又分为nmhaFGF低剂量组(1.25μg组)、nmhaFGF中剂量组(2.5μg组)、nmhaFGF高剂量组(5μg组)各10只,右眼为模型组(给予等量生理盐水)。采用升高眼内压的方法建立大鼠视网膜缺血再灌注模型。再灌注24h后分光光度法测定视网膜MDA、SOD含量,GRIESS反应测定NO含量。结果与模型组相比,nmhaFGF中剂量组、nmhaFGF高剂量组MDA含量显著降低(P<0.01),SOD、NO含量显著升高(P<0.05);nmhaFGF低剂量组和模型组之间SOD、MDA、NO含量没有显著差异(P>0.05)。结论nmhaFGF具有抗视网膜缺血再灌注氧自由基的作用;视网膜缺血再灌注损伤时,nmhaFGF可以增加视网膜内NO的含量,并呈现一定的量效关系。

haFGF和nm-haFGF过表达对乳腺肿瘤细胞增殖及c-fos、c-jun mRNA表达的影响

目的研究非促分裂人酸性成纤维细胞生长因子(non-mitogenetic human acidic fibroblast growth factor,nm-haF-GF)和人酸性成纤维细胞生长因子(haFGF)对恶性肿瘤细胞的增殖作用及其与对c-fos、c-jun mRNA表达的影响,探讨nm-haFGF在治疗神经系统疾病和心血管疾病等方面的临床应用安全性。方法构建细胞内真核表达载体pIRES/haF-GF、pIRES/nm-haFGF,分泌型真核表达载体pSectag/haFGF、pSectag/nm-haFGF。转染乳腺癌细胞MCF-7,Western blot鉴定细胞内及上清中aFGF的表达。用流式细胞技术分析细胞周期(G0/G1,S期,G2/M)。用半定量RT-PCR检测立早基因c-fos、c-jun mRNA的表达。结果nm-haFGF和haFGF在MCF-7细胞中能够过量表达。转染nm-haFGF组的S期和G2/M细胞比率与对照组差异无显著性,而转染haFGF组差异有显著性。转染haFGF后,c-fos、c-jun mRNA的表达明显升高,转染nm-aFGF后却与未转染组无差别。结论与haFGF相比,nm-haFGF消除了促肿瘤细胞增殖活性,基本不会诱导细胞c-fos、c-jun原癌基因的转录与表达。