何首乌饮对衰老大鼠下颌下腺NGF mRNA、EGF mRNA表达的影响

目的探讨何首乌饮对衰老大鼠下颌下腺神经生长因子(NGF)、表皮生长因子(EGF)基因表达的影响。方法选用8周龄SD雌性大鼠90只,用D-半乳糖制造衰老大鼠模型,何首乌饮的干预作用分治疗和预防两部分,用原位杂交法检测各组大鼠下颌下腺NGF mRNA、EGFmRNA表达的情况。结果预防性实验部分的组间NGF mRNA、EGF mRNA表达比较差异有统计学意义(P<0.01),其中以正常组最高,模型组最低,各预防组处于中间水平,其中预防中剂量组最高。治疗性实验部分的组间NGF mRNA、EGF mRNA表达比较差异有统计学意义(P<0.01),其中以阴性对照组最高,自然恢复组最低,各治疗组处于中间水平,其中中剂量组最高。结论何首乌饮可在基因转录水平提高NGF、EGF的表达。     

D-半乳糖诱发的衰老对大鼠下颌下腺端粒酶活性的影响

目的观察D-半乳糖所致衰老大鼠下颌下腺抗氧化能力和端粒酶活性的改变。方法选用8周龄清洁级SD雌性大鼠10只,体重180²20 g,随机分为正常组和模型组,每组5只。采用D-半乳糖诱导大鼠衰老模型,观察下颌下腺谷胱甘肽过氧化物酶(GSH-Px)、总抗氧化能力(T-AOC)改变及采用端粒重复扩增-微孔板杂交法检测下颌下腺细胞端粒酶活性。结果衰老模型组大鼠下颌下腺GSH-Px、T-AOC及端粒酶OD值表达最低,正常组最高(P<0.01)。结论 D-半乳糖所致的大鼠下颌下腺细胞衰老与端粒酶活性降低相关。     

何首乌饮对衰老大鼠下颌下腺NGF、NGF mRNA表达的影响

目的观察何首乌饮对衰老大鼠下颌下腺神经生长因子（NGF）及NGF mRNA表达的影响。方法用D-半乳糖制造衰老大鼠模型,免疫组化及原位杂交法检测何首乌饮预防量、治疗量灌胃后衰老大鼠下颌下腺NGF、NGF mRNA阳性表达的改变。结果预防实验部分NGF、NGF mRNA的表达,模型组与其他各组比较差异均有统计学意义（P（0.05或（0.01）,其中以正常组最高,模型组最低,各预防组中预防中剂量组最高。治疗性实验部分NGF、NGF mRNA的表达,自然恢复组与其他各组比较差异均有统计学意义（P（0.01）,其中以阴性对照组最高,自然恢复组最低,各治疗组中以治疗中剂量组最高。结论何首乌饮可提高衰老大鼠下颌下腺NGF、NGF mR-NA的表达而达到抗衰老的作用。     

何首乌饮对D-半乳糖所致衰老大鼠下颌下腺β-半乳糖苷酶表达的影响

目的 通过观察何首乌饮干预的衰老大鼠下颌下腺β-半乳糖苷酶(β-gal)表达的改变,探讨何首乌饮的抗衰老作用.方法 用D-半乳糖制造衰老大鼠模型,何首乌饮的干预作用分治疗和预防两部分,检测β-半乳糖苷酶活性,观察下颌下腺衰老细胞数目改变.结果 预防性实验部分:模型组衰老细胞数目多于正常组和各预防性用药组;治疗性实验部分:用药各组与自然恢复组相比,衰老细胞数目明显减少,其中治疗中剂量组最低.结论 何首乌饮具有抗模型大鼠下颌下腺细胞衰老的作用.     

何首乌饮对衰老大鼠下颌下腺端粒酶活性的影响

目的观察何首乌饮对衰老大鼠下颌下腺端粒酶活性的影响。方法选用8周龄清洁级SD雌性大鼠20只,随机分为正常组、模型组、何首乌饮预防组、何首乌饮治疗组,每组5只,用D-半乳糖制造衰老大鼠模型,采用端粒重复扩增-微孔板杂交法检测下颌下腺细胞端粒酶活性。结果衰老模型组大鼠下颌下腺端粒酶OD值表达最低,正常组最高,何首乌饮预防组低于正常组,但高于何首乌饮治疗组(P<0.01)。结论何首乌饮可提高衰老大鼠下颌下腺端粒酶活性,且以预防量效果较好。     

何首乌饮对D-半乳糖所致衰老大鼠下颌下腺NGF、EGF表达的影响

目的观察何首乌饮对衰老大鼠下颌下腺神经生长因子(NGF)、表皮生长因子(EGF)表达的影响。方法选用8周龄SD雌性大鼠,随机分为正常组、模型组及何首乌饮干预预防组和治疗组,用SP免疫组化法检测下颌下腺NGF、EGF表达的改变。结果预防性实验各组间NGF、EGF表达均以正常组最高,模型组最低,且中剂量预防组效果最好(P0.01)。治疗性实验各组间NGF、EGF表达均以正常组最高,自然恢复组最低,且中剂量治疗组最高(P0.01)。结论何首乌饮可提高下颌下腺NGF、EGF的表达而达到延缓衰老的作用。     

类似人类老年糖尿病大鼠模型之形态学变化

目的 采用D-半乳糖加高脂高糖乳剂及链脲佐菌素(STZ)构建类似老年人糖尿病的大鼠动物模型,观察其胰腺形态学变化.方法 选用健康Wistar雌性大鼠,20只灌胃生理盐水为正常对照组;20只采用腹腔注射D-半乳糖40 d为衰老模型组;另25只则采用腹腔注射D-半乳糖40 d并灌胃高脂高糖乳剂30 d后腹腔注射STZ(体重≤200 g按30 mg/kg,体重＞200 g者按体表面积进行计算),制备衰老糖尿病模型大鼠,筛选空腹血糖≥11.1 mmol/L者为成模鼠,上述各组大鼠再继续灌胃生理盐水10 d后,各组随机取10个样本以免疫组化、光镜和电镜分别观察大鼠胰岛诱导型一氧化氮合酶(iNOS)表达、胰腺细胞凋亡、胰岛形态及胰岛细胞和腺泡细胞超微结构变化.结果 衰老模型组大鼠胰岛iNOS表达量和胰腺细胞凋亡数较正常对照组增加,衰老糖尿病模型组胰岛iNOS表达量和胰腺细胞凋亡数剧增达3倍以上.HE染色显示,衰老模型组胰岛数量、面积仅为正常的50%～70%,衰老糖尿病模型组胰岛数量、面积仅为正常的20%～30%,且岛内中心部细胞(β细胞)消失尤为显著.电镜观察显示,衰老糖尿病模型组大鼠胰腺细胞明显脱颗粒,呈现膜性结构皱缩,核染色质固缩、碎裂等细胞凋亡征象.结论 采用腹腔注射D-半乳糖加高脂高糖乳剂及腹腔注射STZ,胰腺形态组织学证实大鼠在衰老模型基础上胰岛β、D细胞数量进一步下降可引发糖尿病.     

何首乌饮对D-半乳糖所致衰老大鼠下颌下腺EGF、EGF mRNA表达的影响

目的 观察何首乌饮对衰老大鼠下颌下腺表皮生长因子(EGF)及EGF mRNA表达的影响.方法 采用D-半乳糖衰老大鼠模型,何首乌饮干预分为治疗和预防两部分,用免疫组化及原位杂交法检测下颌下腺EGF、EGF mRNA阳性表达的改变.结果 预防性实验部分EGF、EGF mRNA表达以正常组最高,模型组最低,各预防组中预防中剂量组最高,组间比较有高度统计学意义(P＜0.01).治疗性实验部分以正常组最高,自然恢复组最低,各治疗组中以治疗中剂量组最高,组间比较差异显著(P＜0.01).结论 何首乌饮可提高下颌下腺EGF、EGF mRNA的表达而达到抗衰老的作用.     

类似人类老年糖尿病大鼠模型胰腺形态学变化

目的采用D-半乳糖加高脂高糖乳剂及链脲佐菌素(STZ)构建类似老年人糖尿病的大鼠动物模型,观察其胰腺形态学变化。方法选用健康Wistar雌性大鼠,20只灌胃生理盐水为正常对照组;20只采用腹腔注射D-半乳糖40d为衰老模型组;另25只则采用腹腔注射D-半乳糖40d并灌胃高脂高糖乳剂30d后腹腔注射STZ(体重≤200g按30mg/kg,体重200g者按体表面积进行计算),制备衰老糖尿病模型大鼠,筛选空腹血糖≥11.1mmol/L者为成模鼠,上述各组大鼠再继续灌胃生理盐水10d后,各组随机取10个样本以免疫组化、光镜和电镜分别观察大鼠胰岛诱导型一氧化氮合酶(iNOS)表达、胰腺细胞凋亡、胰岛形态及胰岛细胞和腺泡细胞超微结构变化。结果衰老模型组大鼠胰岛iNOS表达量和胰腺细胞凋亡数较正常对照组增加,衰老糖尿病模型组胰岛iNOS表达量和胰腺细胞凋亡数剧增达3倍以上。HE染色显示,衰老模型组胰岛数量、面积仅为正常的50%～70%,衰老糖尿病模型组胰岛数量、面积仅为正常的20%～30%,且岛内中心部细胞(β细胞)消失尤为显著。电镜观察显示,衰老糖尿病模型组大鼠胰腺细胞明显脱颗粒,呈现膜性结构皱缩,核染色质固缩、碎裂等细胞凋亡征象。结论采用腹腔注射D-半乳糖加高脂高糖乳剂及腹腔注射STZ,胰腺形态组织学证实大鼠在衰老模型基础上胰岛β、D细胞数量进一步下降可引发糖尿病。     

何首乌饮对衰老大鼠下颌下腺凋亡的影响

目的探讨何首乌饮抗下颌下腺衰老的作用。方法分别在预防性和治疗性实验中应用何首乌饮,对D-半乳糖诱发的衰老大鼠模型应用不同剂量何首乌饮。通过TUNEL法检测凋亡细胞,观察下颌下腺凋亡细胞数量。结果预防性实验中下颌下腺细胞凋亡以衰老组最高,正常组最低,预防性应用何首乌饮后以预防中剂量组凋亡最少（P〈0．01）。治疗性实验中下颌下腺细胞凋亡以自然恢复组最高,阴性对照组最低,治疗性应用何首乌饮后以治疗中剂量组凋亡最少（P〈0．01）。结论衰老模型大鼠下颌下腺的凋亡细胞数增多,应用何首乌饮干预后下颌下腺凋亡细胞数减少。何首乌饮可能具有抗下颌下腺衰老的作用。     

哮喘大鼠肾上腺髓质嗜铬细胞表型转化与神经生长因子表达水平初探

目的观察支气管哮喘状态下大鼠肾上腺髓质嗜铬细胞(AMCC)形态和功能的改变以及神经生长因子(NGF)在肾上腺髓质嗜铬细胞中的表达,探讨肾上腺髓质嗜铬细胞与支气管哮喘的关系。方法2005年10月至2006年3月,在中南大学湘雅医院呼吸内科将雄性SD大鼠16只,随机分为对照组和哮喘组,每组8只。哮喘组以鸡卵清蛋白(OVA)致敏和激发哮喘,对照组以生理盐水代替OVA进行致敏和激发。光镜和电镜下观察2组大鼠肾上腺髓质嗜铬细胞的组织结构和超微结构改变,免疫组织化学结合显微图像分析观察肾上腺髓质嗜铬细胞中NGF的表达变化,酶联免疫吸附法(ELISA)检测血清中肾上腺素和去甲肾上腺素的变化。结果HE染色光镜下可见哮喘组大鼠肾上腺髓质嗜铬细胞不同程度的空泡变性样改变,脂质增多,少数可见纤维样物质伸向皮质。电镜下哮喘组大鼠髓质嗜铬细胞线粒体丰富,脂质增多,嗜铬颗粒浓度降低,并且部分出现核膜皱缩现象。与对照组相比,哮喘大鼠NGF免疫阳性反应在肾上腺髓质嗜铬细胞明显上调(P<0.05),ELISA示哮喘组大鼠肾上腺素水平明显低于对照组(P<0.05),去甲肾上腺素水平没有明显变化(P>0.05)。结论支气管哮喘时肾上腺髓质嗜铬细胞NGF的表达增高,增高的NGF可能使肾上腺髓质嗜铬细胞发生表型的转化导致肾上腺素合成与释放不足而参与哮喘的发生、发展。     

清脑益元汤对大鼠脑缺血损伤Nestin、NGF表达的影响

目的研究清脑益元汤对大鼠脑缺血损伤后巢蛋白(Nestin)、神经生长因子(NGF)表达的影响,探究清脑益元汤对缺血性脑损伤神经元保护作用的部分机制。方法将192只健康SD大鼠随机分为两大组,每大组再分为4个亚组:空白组(n=6)、假手术组(n=30)、模型组(n=30)、清脑益元汤组(n=30)。采用改良的Longa线栓法制备大鼠急性脑缺血损伤模型,除空白组外,假手术组、模型组及清脑益元组按缺血损伤后1 d、3 d、7 d、14 d、28 d 5个取材时间点分为5个亚组。造模成功后取大鼠缺血侧脑组织,采用免疫组化法检测大鼠缺血侧皮质区Nestin、NGF表达,采用实时荧光定量PCR(Real-Time PCR)法检测大鼠缺血侧皮质区Nestin、NGF mRNA表达。结果(1)免疫组化法:与假手术组相比,模型组、清脑益元组在缺血侧Nestin、NGF阳性细胞明显增多(P<0.01,P<0.01);与模型组相比,清脑益元组Nestin、NGF阳性细胞表达明显增多(P<0.01,P<0.05);(2)Real-Time PCR法:与假手术组相比,模型组、清脑益元汤组大鼠脑组织缺血侧皮质区Nestin、NGF mRNA表达量增加(P<0.01,P<0.01);与模型组相比,清脑益元汤组大鼠脑组织缺血侧皮质区Nestin、NGF mRNA表达量升高(P<0.01,P<0.05)。结论清脑益元汤可能通过上调脑缺血损伤后Nestin、NGF蛋白及mRNA表达,促进脑组织损伤修复、保护神经元,进而发挥脑保护的作用。     

环磷酸腺苷-蛋白酶A信号系统参与神经生长因子促进脑缺血再灌注大鼠轴突再生

目的观察脑室注射神经生长因子(NGF)对脑缺血再灌注大鼠神经再生的影响及其与环磷酸腺苷-蛋白激酶A(cAMP-PKA)信号途径的关系。方法采用线栓法制作大鼠脑缺血再灌注模型,将60只大鼠分为假手术组(对照组),单纯脑缺血再灌注组(缺血组),NGF1组,NGF2组(脑缺血再灌注并脑室注射NGF,分别注入0.5及2.5μg/100 g体重)。用放免法检测缺血周边区脑组织cAMP的含量,逆转录聚合酶链反应的方法检测生长相关蛋白43(GAP-43)及蛋白激酶A(PKA)mRNA的表达。结果缺血组6、24 h cAMP的含量下降,GAP-43及PKA mRNA表达减少,NGF治疗组脑组织GAP-43 mRNA表达较缺血组增加,且这种变化与cAMP的含量及PKA mRNA表达增加相一致,并呈剂量依赖性。结论NGF能够促进脑缺血再灌注后的轴突再生,cAMP-PKA信号途径在这个过程中起重要作用。     

Serum EGF and NGF levels of patients with brain injury and limb fracture

ObjectiveTo explore the expression and significance of human epidermal growth factor (EGF) and nerve growth factor (NGF) in patients with knee osteoarthritis.MethodsRT-PCR and enzyme-linked immunosorbent assay were used to measure the serum EGF and NGF expression levels of patients with limb fracture and brain trauma injurry after 1 d, 3 d, 7 d, 14 d and the relationship between them was analyzed. The level was compared among the simple fracture group, traumatic brain injury group and the normal control group, with 40 cases in each group.ResultsThe serum NGF levels were significantly different among three groups. Serum NGF, EGF mRNA and protein levels gradually decreased with the increasing injury time in the limb fracture combined with brain injury group, traumatic brain injury group, the simple fracture group and the health control group (P<0.05).ConclusionsThe serum of NGF, EGF levels significantly increased when limb fracture combined with brain injury, so EGF and NGF may be involved in the process of fracture healing.     

Lack of effect of antipsychotics on BNDF and NGF levels in hippocampus of Wistar rats

Schizophrenia is a common and serious mental disorder, in which the majority of patients require long-term antipsychotic treatment. Several studies have suggested that schizophrenia is associated with decreased neurotrophins such as brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF). Investigation of the mechanisms of pharmacological agents that are used in the treatment of schizophrenia has been used to better understand the basis of the pathology associated with this mental illness. The present study aims to investigate the effect of chronic treatment with antipsychotics, named haloperidol (HAL), clozapine (CLO), olanzapine (OLZ) or aripiprazole (ARI) on BDNF and NGF levels in rat hippocampus. Adult male Wistar rats received daily injections of HAL (1.5 mg/kg), CLO (25 mg/kg), OLZ (2.5, 5 or 10 mg/kg) or ARI (2, 10 or 20 mg/kg), whereas control animals were given vehicle. BDNF and NGF levels were measured in rat hippocampus by sandwich-ELISA. The results showed that chronic administration of antipsychotics did not modify BDNF and NGF levels in rat hippocampus, suggesting that their therapeutic properties are not mediated by stimulation of these neurotrophins.     

Age-related regional differences in cardiac nerve growth factor expression

Age has been identified as an independent risk factor for cardiovascular diseases. A shift of the cardiac autonomic nervous system towards an increase in sympathetic tone has been reported in the elderly. Nerve growth factor (NGF) is the main neurotrophic factor that increases the sympathetic activity of the heart. If there is a shift of NGF expression in old compared to young cardiomyocytes and whether there are regional differences in the heart still remain unclear. Therefore, we chose a rat model of different-aged rats (3–4 days = neonatal, 6–8 weeks = young, 20–24 months = old), and isolated cardiomyocytes from the left and the right atrium (LA, RA), as well as from the left and the right ventricle (LV, RV), were used to determine NGF expression on mRNA and protein levels. In neonatal, young, and old rats, NGF amount in LA and RA was significantly lower as compared to LV and RV. In young and old rats, we found significant higher NGF protein levels in LA compared to RA. In addition, both atria showed an increase in NGF expression between age groups neonatal, young, and old. In both ventricles, we observed a significant decrease in NGF expression from neonatal to young rats and a significant increase from young to old rats. The highest NGF amount in LV and RV was observed in neonatal rats. Regarding tyrosine kinase A receptor (TrkA) expression, the main receptor for NGF signaling, both atria showed the largest expression in old rats; while in LV and RV, TrkA was expressed mainly in young rats. These results point to a contribution of nerve growth factors to the change of autonomic tone observed in elderly patients.     

Role of IL-1b and TNF-a in the regulation of NGF in experimentally induced arthritis in mice

The aim of this study was to investigate the effects of IL-1β and TNF-α on NGF levels in the knee joint in experimentally induced arthritis in adult mice. Out data showed that IL-1β, but not TNF-α, induces an increase in NGF levels, while concomitant injection of both cytokines enhances the effect of IL-1β on NGF presence. Analysis of NGF levels in the knee joint of carrageenan- and IL-1β-induced inflammation after administration of antibodies against TNF-α supports this hypothesis. Our studies also showed that exogenous administration of NGF antibody reduces the enhanced level of TNF-α occurring in arthritic mice. This latter observation indirectly suggests that NGF is implicated in the upregulation of TNF-α in these animal models of joint inflammation. The functional significance of NGF or NGF antibody in inflammatory arthritis is discussed. Received: 16 March 1998 / Accepted: 2 July 1998     

Studies on the expression of the beta nerve growth factor (NGF) gene in the central nervous system: level and regional distribution of NGF mRNA suggest that NGF functions as a trophic factor for several distinct populations of neurons.

Beta nerve growth factor (NGF), a target-derived protein necessary for survival and development of sympathetic and sensory neurons, can also affect subpopulations of neurons in the central nervous system (CNS). Using a blot hybridization assay capable of detecting 10 fg of mRNA, we measured the levels of NGF mRNA in the major brain regions, including those innervated by NGF-responsive neurons. NGF mRNA was detected unambiguously in each major region of the CNS. The levels were comparable to those in sympathetic effector organs. Discrete areas contained very different amounts of NGF mRNA. Up to 40-fold differences were seen, a range comparable to the differences between richly and sparsely innervated sympathetic effector organs. The highest concentrations of NGF mRNA were found in the cortex and hippocampus, which are the major targets of the NGF-responsive cholinergic neurons of the basal forebrain nuclei. Significant amounts of NGF mRNA were also found in areas that contain the central processes of NGF-responsive sensory neurons, such as the pons, medulla, and spinal cord. The presence of NGF mRNA in these areas suggests that brain NGF may act as a target-derived trophic factor for both populations of neurons. NGF mRNA was also found in the striatum, suggesting that locally derived NGF may act there as a trophic factor for a recently identified population of NGF-responsive cholinergic local circuit neurons. However, high levels of NGF mRNA were also found in some regions, such as the diencephalon, that have no relation to any identified population of NGF-responsive neurons. This suggests that there may be additional populations of NGF-responsive neurons in the CNS that have not yet been discovered.     

大鼠骨髓基质干细胞体外分化为神经元样细胞并表达肠神经相关神经营养因子

目的探讨维甲酸、锌联合胎肠培养基诱导大鼠骨髓基质干细胞向神经元细胞分化的可能性以及分化前后肠神经相关神经营养因子表达的变化。方法体外培养大鼠骨髓基质干细胞（BMSC），传代至第6代，进行神经诱导分化。先以bFGF（10ng／ml）预诱导24h，然后以维甲酸（Retinoic acid，RA）、锌在胎肠培养基（FGCM）条件下诱导10d。免疫荧光法检测神经元标志神经特异性烯醇化酶（NSE）和神经丝蛋白（NF）的表达。RT-PCR法检测胶质源神经营养因子（GDNF）和神经生长因子（NGF）mRNA的表达。结果流式细胞检测BMSC高表达CD90（99，7％），而CD45表达阴性。诱导10d后，部分细胞在形态上表现出神经元样改变，免疫荧光染色神经元标志NSE和NF阳性，而胶质细胞标志GFAP阴性。RT-PCR检测发现，BMSC诱导前低表达神经营养因子NGF和GDNF mRNA，而诱导后表达水平显著增加。结论维甲酸、锌联合胎肠培养基不仅能在体外诱导BMSC分化为神经元样细胞，而且能促进肠神经相关神经营养因子表达显著上调。