碱性成纤维细胞生长因子和血管内皮生长因子在糖尿病骨骼肌病变中的变化及意义

目的 探讨糖尿病骨骼肌碱性成纤维细胞生长因子（bFGF）及血管内皮生长因子（VEGF）表达的变化及其在糖尿病骨骼肌病变中的作用。方法 选用Wistar大鼠，随机分成4组：正常对照组、单纯后肢缺血组、糖尿病组和糖尿病后肢缺血组。以四氧嘧啶50mg／kg尾静脉注射制造糖尿病模型。糖尿病造模后2周结扎右股动脉制造缺血模型。10周后，观察腓肠肌病理形态学改变，同时采用免疫组化方法检测bFGF和VEGF蛋白表达情况。结果 糖尿病组骨骼肌表现为普遍性肌萎缩，肌纤维变性坏死。单纯缺血组和糖尿病组bFGF蛋白表达减弱（表达阳性率分别为53．66％和35．51％），糖尿病后肢缺血组bFGF的阳性率为25．42％，下降明显（P〈0．01）。糖尿病缺血组VEGF表达阳性率为20．68％，低于单纯缺血组（54．04％，P〈0．01）。结论 高血糖和缺血引起肌肉中bFGF及VEGF等生长因子的变化可能是糖尿病骨骼肌萎缩的重要原因之一。     

皮肤血管瘤患儿脉冲激光治疗前后外周血生长因子的变化

目的 对比分析皮肤血管瘤患儿脉冲激光治疗前后外周血血管内皮生长因子(VEGF)、转化生长因子(TGF-β1)及碱性成纤维细胞生长因子(bFGF)的表达变化.方法 选取皮肤血管瘤患儿73例(增生期35例、消退期17例、消退完成期21例)及非血管性皮肤病患儿22例(对照组),所有增生期皮肤血管瘤患儿均采用脉冲染料激光进行治疗;取外周静脉血,检测VEGF、TGF-β1及bFGF的mRNA表达水平及蛋白含量.结果 治疗前,增生期患儿的VEGF、TGF-β1及bFGF的mRNA表达水平和蛋白含量高于消退期,消退期高于消退完成期与对照组,差异均有统计学意义(P＜0.05),消退完成期和对照组之间无统计学差异(P＞0.05).治疗后,增生期患儿VEGF、TGF-β1及bFGF的mRNA表达水平和蛋白含量均显著低于治疗前(P＜0.05).结论 脉冲染料激光可下调皮肤血管瘤患儿外周血VEGF、TGF-β1及bFGF的mRNA表达水平,从而减少各蛋白分泌,可能为其发挥治疗作用的重要机制之一.     

bFGF联合骨髓干细胞动员剂促进大鼠缺血后肢血管新生的分析

目的：了解bFGF联合骨髓干细胞动员剂促进大鼠缺血后肢血管新生的作用。方法：将48只大鼠的左侧股动脉与其分支切断,构建大鼠后肢缺血模型,并按随机数字表法分成四组,每组12只,其中A组将重组人bFGF蛋白注入大鼠缺血后肢肌内;B组将重组人粒细胞集落刺激因子（rhG-CSF）注入大鼠皮下;C组大鼠予以bFGF联合rhG-CSF;D组予以均等的生理盐水。治疗持续4周后予以每组大鼠腹主动脉造影,进而观察大鼠侧支血管的具体形成状况,予以内收肌以及腓肠肌肌组织病理切片检查,借助图像分析系统,统计各组大鼠血管密度。行免疫组化检测,分析大鼠血管内皮生长因子阳性表述血管数。结果：在大鼠侧支循环血管条数、血管内皮生长因子阳性表述血管数以及血管密度上,C组〉A组〉B组〉D组,差异均有统计学意义（P〈0.05）。结论：对于促大鼠缺血后肢血管新生而言,bFGF蛋白与骨髓干细胞动员剂疗效显著,均可改善大鼠肢体的缺血状态,但骨髓干细胞动员剂联合bFGF蛋白改善大鼠缺血肢体更为明显。     

培哚普利对急性心肌梗死患者血管内皮功能及EPCs水平的干预作用

目的探讨培哚普利对急性心肌梗死(AMI)的疗效及对患者血管内皮功能和内皮祖细胞(EPCs)水平的影响。方法选取2016年1月~2018年6月本院收治的94例AMI患者为研究对象,将所有患者按照治疗方案分为观察组(n=47)及对照组(n=47),2组均行急诊经皮冠状动脉介入治疗,对照组术后1 d给予常规基础治疗,观察组同时给予培哚普利叔丁胺片治疗,逐渐加量至8 mg/d,2组均持续治疗6个月。于治疗前后检测并比较2组患者血压、血钾、心率(HR)及超敏C-反应蛋白(hs-CRP)、血管紧张素Ⅱ(Ang II)、血管内皮生长因子(VEGF)等指标变化;检测并比较2组治疗前后肱动脉内皮依赖性血管舒张功能(FMD)及EPCs数量。结果与治疗前比较,治疗后2组血浆hs-CRP水平及观察组SBP、DBP、Ang II水平明显降低(P0.05或P0.01)。结论培哚普利可显著降低AMI患者血压、炎性因子水平,动员EPCs细胞,进而改善血管内皮功能,且不会影响患者血钾及HR等,具有一定的安全性。     

bFGF对大鼠缺血性脑损伤CaMKⅡmRNA表达影响

目的 研究大鼠脑缺血再灌注后Ca2 >/钙调蛋白依赖的蛋白激酶Ⅱ(CaMK(Ⅱ)mRNA的表达,探讨碱性成纤维细胞生长因子(bFGF)对脑组织缺血再灌注神经元CaMK Ⅱ mRNA的调节作用及机制.方法 应用线栓法制作大鼠局灶性脑缺血再灌注模型,大脑中动脉阻塞1 h再灌注损伤24 h,采用TUNEL法、逆转录聚合酶链反应(RT-PCR)检测海马及皮质内神经元凋亡和CaMK Ⅱ mRNA的表达.结果 大鼠缺血再灌注海马及顶叶皮质内神经元凋亡数增加,而CaMKⅡ mRNA表达较假手术组减少,注射bFGF后神经元凋亡数减少,CaMK Ⅱ mRNA表达明显高于缺血再灌注组.结论 bFGF抑制缺血神经元凋亡,参与CaMK Ⅱ mRNA的调节,对缺血神经元有保护作用.     

bFGF对脑缺血大鼠脑组织HSP70 mRNA表达影响

目的 研究大鼠脑缺血再灌注后热休克蛋白70(HSP70)mRNA的表达,探讨碱性成纤维细胞生长因子(bFGF)对脑组织缺血再灌注神经元HSP70 mRNA的调节作用及机制.方法 应用线栓法制作大鼠局灶性脑缺血再灌注模型,大脑中动脉阻塞1h再灌注损伤24h,采用原位缺口末端标记法(TUNEL)、RT-PCR法检测海马及皮质内神经元凋亡和HSP70 mRNA的表达.结果 大鼠缺血再灌注海马及顶叶皮质内神经元凋亡数增加而HSP70mRNA表达较正常组增加.注射bFGF后神经元凋亡数减少,HSP70mRNA表达明显高于缺血再灌注组.结论 bFGF抑制缺血神经元凋亡,参与HSP70 mRNA的调节,对缺血神经元有保护作用.     

柴朴汤下调哮喘大鼠血管内皮生长因子表达抑制气道的重塑

目的观察血管内皮生长因子(VEGF)在慢性哮喘大鼠中的表达水平及柴朴汤的干预作用,探讨柴朴汤在气道重塑中的作用。方法以卵清白蛋白致敏及反复激发建立大鼠慢性哮喘模型,常规切片观察气管内管壁厚度及气道平滑肌厚度。免疫组化及逆转录-聚合酶链方法检测肺内VEGF蛋白和mRNA的表达。结果慢性哮喘模型组大鼠气管内壁及气道平滑肌增厚,肺组织VEGFmRNA、VEGF蛋白的表达增高(P<0.01)。结论VEGF参与了慢性哮喘大鼠的气道重塑。柴朴汤能够抑制气道重塑。     

糖末宁颗粒对糖尿病周围神经病变大鼠坐骨神经血管内皮生长因子表达的影响

目的 探讨糖末宁颗粒对糖尿病周围神经病变(DPN)大鼠坐骨神经血管内皮生长因子(VEGF)蛋白表达的影响.方法 将32只Wisar大鼠随机分为正常对照组、模型组、糖末宁组,采用2％链脲佐霉素(STZ)53 mg/kg,诱导糖尿病大鼠模型.实验期间每日灌服糖末宁混悬液,实验结束后以透射电镜观察大鼠坐骨神经超微结构;应用western blot及免疫组化技术检测各组大鼠坐骨神经VEGF蛋白表达的情况.结果 坐骨神经VEGF蛋白表达,模型组大鼠较正常对照组增高,糖末宁组大鼠低于模型组,但较正常对照组略高.结论 糖末宁颗粒可抑制DPN大鼠坐骨神经VEGF蛋白表达,对神经损害有一定的保护作用.     

VEGF对缺血/再灌注损伤胰腺细胞凋亡的影响

目的 研究血管内皮生长因子(VEGF)对缺血/再灌注损伤胰腺组织细胞凋亡的影响.方法 将雄性sD大鼠30只随机分为3组(n=10),A组为假手术组,B组为缺血/再灌注损伤组,C组为缺血/再灌注损伤+VEGF反义寡核苷酸组.通过血管夹阻断大鼠腹腔干及肠系膜上动脉30 min,然后去除血管夹再灌注6 h,建立大鼠胰腺缺血/再灌注损伤模型.对各组胰腺组织进行VEGF免疫组化染色及TUNEL法细胞凋亡检测.结果 缺血/再灌注损伤后胰腺组织出现细胞凋亡,同时VEGF蛋白表达上调.缺血/再灌注损伤+VEGF反义寡核苷酸组的胰腺组织VEGF蛋白表达较缺血/再灌注损伤组显著减少(P<0.05),前者细胞凋亡指数较后者明显升高(P<0.05).结论 VEGF能抑制缺血/再灌注损伤胰腺细胞凋亡,可能对胰腺缺血再灌注损伤具有保护作用.     

人肺腺癌细胞缺氧后血管形成诱导活性的变化

目的 探讨肺腺癌细胞经过缺氧刺激后的血管形成诱导活性的变化。方法 将人肺腺癌细胞株A549暴露于常氧、缺氧和无氧环境48h后，将其条件培养基或癌细胞分别与包含在纤维蛋白胶中的内皮细胞团块共培养，计数从内皮细胞团块长出的小管数量以反映血管形成诱导活性。同时通过ELISA法分析条件培养基和两细胞共培养时的培养基中的血管内皮细胞生长因子(VEGF)、碱性成纤维细胞生长因子(bFGF)含量。结果 缺氧组条件培养基以及共培养时的血管形成诱导活性及培养基中VEGF、bFGF水平均显著高于常氧组。无氧组血管形成诱导活性及两种生长因子水平均显著低于常氧组。结论 适度缺氧可以刺激人肺腺癌细胞VEGF、bFGF等生长因子的表达，增强血管形成诱导活性。但是严重缺氧可削弱VEGF、bFGF表达，减弱癌细胞的血管形成诱导活性。     

Investigation of ocular neovascularization-related genes and oxidative stress in diabetic rat eye tissues after resveratrol treatment

Changes in vascular endothelial growth factor (VEGF), angiotensin-converting enzyme (ACE), matrix metalloproteinase (MMP)-9, and endothelial nitric oxide synthase (eNOS) mRNA expression profiles and oxidative stress in the eye tissue microenviroment may have important roles in ocular neovascularization and permeability in proliferative diabetic retinopathy. The present study investigated the effects of resveratrol (RSV) treatment on the mRNA expression profile of VEGF, ACE, MMP-9, and eNOS, which are associated with vascular neovascularization, and glutathione, protein carbonyl, and nitrite-nitrate levels, which are markers of oxidative stress in eyes of diabetic rats. Twenty-four Wistar albino male rats were divided into four groups. After diabetes induction with streptozotocin (10?mg/kg/day) RSV was administered to the RSV and diabetes mellitus (DM)?+?RSV groups for 4 weeks. The mRNA levels were measured by quantitative real-time polymerase chain reaction assay, and biochemical estimations were determined with spectrophotometric assays in eye homogenates. The mRNA expression levels of VEGF, ACE, and MMP-9 were increased in the DM group compared with the control group, and RSV treatment decreased their mRNA levels. Expression of eNOS mRNA was increased in the RSV and DM groups and decreased in the DM?+?RSV group. Nitrite-nitrate levels and protein carbonyl content were increased and glutathione levels were decreased in the DM group compared with controls. Consequently, these data suggest that RSV suppressed the expression of eNOS, which is actively involved in the inflammation and healing process in chronic diabetes. Although oxidative stress was increased in eye tissue from diabetic rats, mRNA levels of VEGF, MMP-9, and ACE genes associated with vascular remodeling did not change in diabetic eyes.     

牛磺酸对糖尿病大鼠视网膜超微结构的影响

目的 观察糖尿病大鼠视网膜早期病变，探讨牛磺酸对糖尿病大鼠视网膜超微结构的影响。方法 选择健康成年SD大鼠50只，随机分成正常对照组(CON)、糖尿病1月组(DM1)、牛磺酸干预的糖尿病1月组(DM1 T)、糖尿病3月组(DM3)、牛磺酸干预的糖尿病3月组(DM3 T)。一次性腹腔内注射链尿佐菌素(STZ)，建立糖尿病大鼠模型。在普通饲料中加入1.2％牛磺酸进行营养干预，对各组视网膜进行光镜HE染色及透射电镜观察。结果 视网膜HE染色显示，糖尿病组视网膜较正常对照组明显变薄，神经节细胞数明显减少。牛磺酸干预后视网膜厚度较糖尿病组增加，神经节细胞数相对增多。观察超微结构发现糖尿病大鼠感光细胞外段膜盘排列紊乱，膜盘间隙增大，严重者出现膜盘断裂、溶解；视杆内段线粒体等细胞器肿胀变性，内网状层轴突肿胀、空泡样变。牛磺酸干预后感光细胞外段膜盘排列明显变整齐，线粒体肿胀较轻，内网状层轴突肿胀较糖尿病组轻。结论 牛磺酸可明显改善糖尿病视网膜的超微结构改变。     

Vascular endothelial growth factor gene expression in patients' ischemic skeletal muscle with diabetic foot

OBJECTIVE: To investigate the relationship between the endogenous vascular endothelial growth factor (VEGF) gene expression in diabetics' calf ischemic skeletal muscle and the pathogeny of diabetic foot. METHODS: Twenty-four patients (33 limbs) were divided into 3 groups: diabetes mellitus (DM) without lower extremity ischemia (n = 5) (10 limbs); arteriosclerosis obliterans (ASO) without diabetes mellitus (n = 10) (13 limbs); diabetic lower limb arteriosclerosis obliterans (DLASO) (n = 9) (10 limbs). Control group consisted of normal volunteers (NOR) (n = 5) (10 limbs). The calf skeletal muscle tissue was obtained through muscle biopsy. RT-PCR was applied to determine the expression of hVEGF165mRNA. RESULTS: There was no expression in the calf skeletal muscle tissue of normal volunteers and DM. The calf skeletal muscle tissue in DLASO had the expression of hVEGF165mRNA (0.021 +/- 0.013) micro g, but obviously lower than ASO (0.133 +/- 0.024) micro g, (t = 13.32, P < 0.01). CONCLUSIONS: The endogenous VEGF gene expression in ischemic lower extremity of DLASO is obviously lower than that of ASO. It is the important endogenic cause of the genesis and development of diabetic foot ulcer.     

VEGF对缺血脑组织表达Fas/Fasl的影响

目的探讨VEGF治疗脑梗死的机制.方法用线拴法制成Wistar大鼠大脑中动脉永久性闭塞模型,将VEGF165真核表达质粒pUCCAGGS/hVEGF165经颅骨注入到缺血区.术后7d断头取脑,用逆转录PCR(RT-PCR)检测VEGF mRNA的表达强度,用免疫组织化学方法检测Fas、Fasl和VEGF的表达水平.结果与对照组相比,治疗组VEGFmRNA的表达增强,VEGF表达增高(P＜0.01),Fas、Fasl表达减弱(P＜0.01).结论VEGF165基因可以转化到缺血脑组织中并表达VEGF mRNA和VEGF,后者可能通过抑制Fas和Fasl的表达而保护神经细胞.     

L-Carnitine changes the levels of insulin-like growth factors (IGFs) and IGF binding proteins in streptozotocin-induced diabetic rat

This study investigated the effects of L-carnitine on insulin-like growth factor-I/II (IGF-I/II) and insulin-like growth factor binding proteins (IGFBPs) in streptozotocin (STZ)-induced diabetic rats. Each rat in the three L-carnitine-treated groups was injected subcutaneously with L-carnitine, 50 (D50), 100 (D100), or 200 (D200) mg/kg body weight every other day for four weeks, and animals in normal (N) and diabetic (DM) groups received saline by the same method. Diabetic rats had significantly lower carnitine concentrations in serum and liver compared with normal rats. Total carnitine concentrations were increased dose-dependently by carnitine treatment. Total IGF-I in serum from diabetic rats was increased dose-dependently by carnitine treatment, but was statistically significant only in the D200 group. The expression of liver IGF-I mRNA was lower in diabetic rats than in normal rats and increased by L-carnitine treatment. L-Carnitine treatment of diabetic rats had no effect on the levels of IGF-II in serum, liver, and kidney. Although the levels of IGF-II in serum and kidney of diabetic rats were increased in comparison with normal rats, IGF-II mRNA was not expressed in liver. Diabetic rats had markedly lower IGFBP-3 than normal rats did, and IGFBP-3 was increased by L-carnitine treatment. These results demonstrate that L-carnitine treatment of diabetic rats modulates the IGFs/IGFBPs axis. Especially note-worthy is that L-carnitine at a dose of 200 mg/kg/48 h for four weeks was able to restore serum total IGF-I in STZ-induced diabetic rats to nearly normal levels.     

三种食物多酚对化学诱导DM大鼠糖脂代谢的影响

目的探讨膳食多酚[绿原酸、表没食子儿茶素没食子酸酯(EGCG)、槲皮素]对链脲佐菌素(STZ)诱导的糖尿病大鼠血糖、血脂、肝脏中葡萄糖-6-磷酸酶(G-6-pase)和骨胳肌组织中葡萄糖转运体4(GLUT4)表达的影响。方法单次腹腔注射链脲佐菌素(STZ,35 mg/kg)结合高脂饮食建立糖尿病大鼠模型,将成模大鼠分成5组[糖尿病模型组(DM)、糖尿病模型+二甲双胍组(S)、糖尿病模型+绿原酸组(CA)、糖尿病模型+EGCG组(E)、糖尿病模型+槲皮素组(Q)],另设正常对照组(NC),分别灌喂二甲双胍、绿原酸、EGCG和槲皮素4w后,测定其糖耐量、空腹胰岛素、甘油三酯、胆固醇、G-6-pase和GLUT4 mRNA的表达。结果绿原酸、EGCG、槲皮素均表现出改善STZ诱导的糖尿病大鼠血糖、甘油三酯(TG)和胆固醇(TC)的含量,并能改善糖尿病模型大鼠的胰岛素敏感性,抑制肝脏G-6-pase mRNA的表达,且提高了骨胳肌GLUT4 mRNA的表达,其中以绿原酸效果最佳。仅仅其糖耐量改善弱于槲皮素作用,但均弱于阳性对照组二甲双胍的作用。结论绿原酸、EGCG、槲皮素均能有效改善STZ诱导的SD糖尿病大鼠的糖代射、脂代射、胰岛素敏感生及肝脏G-6-pase mRNA和骨胳肌GLUT4 mRNA的表达,绿原酸的效果最佳。[营养学报,2012,34(6):572-575,581]     

铬对糖尿病大鼠骨骼肌组织基因表达的调控作用

目的　研究铬对糖尿病大鼠代谢相关基因表达的调控作用。方法　雄性Wistar大鼠随机分成 3组 :正常对照组、糖尿病对照组、糖尿病补铬组。糖尿病补铬组每日以 2 0 0 μg kgbw的铬连续灌胃 6 0天。采用mRNA差异显示技术、基因片段的克隆、测序、同源性分析等观察各组大鼠之间骨骼肌中基因表达的变化。结果　分离到糖尿病补铬组与糖尿病对照组之间出现明显差异的 4 0 0bp以上的cDNA片段 11条 ,其中在糖尿病补铬组中高表达的cDNA片段 4条 ;在糖尿病对照组中高表达的cDNA片段 7条。同源性比较结果显示Cr 3与GLUT4 (葡萄糖转运体 4 )的同源性为 98% ;Cr 5与IRS 1(胰岛素受体底物 1)的同源性为 10 0 % ,RT PCR验证结果显示GLUT4表达量在糖尿病补铬组高于糖尿病对照组。结论　铬可诱导糖尿病大鼠骨骼肌中GLUT4mRMA的表达 ,这可能是铬改善糖尿病糖、脂代谢紊乱的分子机制之一。     

Effects of glutamine supplementation on oxidative stress-related gene expression and antioxidant properties in rats with streptozotocin-induced type 2 diabetes

There are close links among hyperglycaemia, oxidative stress and diabetic complications. Glutamine (GLN) is an amino acid with immunomodulatory properties. The present study investigated the effect of dietary GLN on oxidative stress-relative gene expressions and tissue oxidative damage in diabetes. There were one normal control (NC) and two diabetic groups in the present study. Diabetes was induced by an intraperitoneal injection of nicotinamide followed by streptozotocin (STZ). Rats in the NC group were fed a regular chow diet. In the two diabetic groups, one group (diabetes mellitus, DM) was fed a common semi-purified diet while the other group received a diet in which part of the casein was replaced by GLN (DM-GLN). GLN provided 25% of total amino acid N. The experimental groups were fed the respective diets for 8 weeks, and then the rats were killed for further analysis. The results showed that blood thioredoxin-interacting protein (Txnip) mRNA expression in the diabetic groups was higher than that in the NC group. Compared with the DM group, the DM-GLN group had lower glutamine fructose-6-phosphate transaminase 1, a receptor of advanced glycation end products, and Txnip gene expressions in blood mononuclear cells. The total antioxidant capacity was lower and antioxidant enzyme activities were altered by the diabetic condition. GLN supplementation increased antioxidant capacity and normalised antioxidant enzyme activities. Also, the renal nitrotyrosine level and Txnip mRNA expression were lower when GLN was administered. These results suggest that dietary GLN supplementation decreases oxidative stress-related gene expression, increases the antioxidant potential and may consequently attenuate renal oxidative damage in rats with STZ-induced diabetes.