槲皮素对D-半乳糖致衰老小鼠模型学习记忆及脑内炎症的影响

目的探讨槲皮素对D-半乳糖致衰老小鼠模型学习记忆能力及脑内炎症通路的影响。方法将昆明小鼠32只随机分为对照组(NC组)、模型组(M组)、Q1治疗组(Q1组)、Q2治疗组(Q2组),每组8只。M组、Q1组及Q2组小鼠皮下注射D-半乳糖100 mg/(kg·d)建立小鼠衰老模型,NC组给予等量生理盐水。Q1、Q2组造模同时给予不同剂量槲皮素5、10 mg/(kg·d)灌胃干预,NC组和M组给予相同剂量生理盐水灌胃处理,干预8周。Morris水迷宫评价逃避潜伏期、穿越平台次数及平台象限滞留时间,Western blotting检测小鼠脑内高级糖基化终末产物(AGEs)及其受体(RAGE)、核转录因子(NF-κB)和其下游炎症因子肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、环氧化酶-2(COX-2)表达水平。采用SPSS 20.0统计软件对数据进行分析。组间比较采用完全随机设计的单因素方差分析或t检验。结果与NC组比较,M组小鼠逃避潜伏期延长、穿越平台次数及在平台所在象限滞留时间降低,脑内AGEs、RAGE、NF-κB及炎症因子TNF-α、IL-1β、COX-2水平显著升高(P0.05)。与M组比较,治疗组小鼠逃避潜伏期缩短、穿越平台次数及在平台所在象限滞留时间延长,脑内炎性因子水平减少(P0.05)。与Q1组比较,Q2组小鼠逃避潜伏期缩短、穿越平台次数及在平台所在象限滞留时间延长,RAGE、IL-1β和COX-2表达水平显著降低。结论槲皮素能够有效改善衰老小鼠学习记忆功能障碍,抑制AGEs/RAGE/NF-κB炎症通路,降低脑内炎性因子的生成,具有抗衰老潜力。     

鸡冠花对D-半乳糖致小鼠衰老作用的研究

目的　研究鸡冠花的抗衰老作用。方法　随机将小鼠分为 A(正常对照组 )、B(衰老对照组 )、C(鸡冠花高剂量组 ) D(鸡冠花低剂量组 )四组。每组 1 4只 ,雌雄各半 ,除 A组外 ,其余三组每天注射 0 .1 g/ kg D-半乳糖 ,建立衰老动物模型 ,C、D两组同时施加不同剂量鸡冠花 ,连续 8w,测定各组血清超氧化物歧化酶 ( SOD)、谷胱甘肽过氧化物酶 ( GSH- Px)活性 ,总抗氧化能力 ( T- AOC) ,丙二醛 ( MDA)和肝脏脂褐质 ( L F )含量。结果　衰老对照组 SOD、GSH- Px活性和 T- AOC较正常对照组明显降低 ( P<0 .0 5) ,MDA和 L F含量增加 ( P<0 .0 5) ,呈现衰老状态。鸡冠花两组 SOD、GSH- Px活性和 T- AOC较衰老对照组明显升高 ( P<0 .0 5) ,MDA和 L F含量减少 ( P<0 .0 5)。结论　鸡冠花具有拮抗 D-半乳糖致衰老作用     

卷丹水提物对D-半乳糖衰老模型小鼠的影响

目的观察卷丹水提物对D-半乳糖衰老模型小鼠的影响。方法建立D-半乳糖致小鼠衰老模型，以Y型电迷宫检测其学习记忆能力，分别采用黄嘌呤氧化酶法、TBA比色法测定大脑SOD活性和MDA含量，观察小鼠服用卷丹水提物后上述指标的变化。结果与D-半乳糖模型组相比，卷丹水提物低、中、高剂量均可显著提高衰老模型小鼠的学习记忆能力（P〈0．05或P〈0．01），显著增加D-半乳糖所致衰老模型小鼠大脑SOD活性、降低MDA含量（P〈0．05）。结论卷丹水提物能改善D-半乳糖所致衰老小鼠的多项体征。     

迷迭香酸对D-半乳糖致衰老小鼠抗氧化作用的研究

目的 观察迷迭香酸对D-半乳糖诱导的衰老小鼠的抗氧化作用.方法 昆明小鼠连续6 w颈背部皮下注射D-半乳糖1mg·g-1·d-1复制亚急性衰老小鼠模型.造模的过程中,3个给药组(低、中、高剂量) 分别以100、200和400 mg·kg-1·d-1的迷迭香酸分析纯灌胃;同时设立3个含有等效剂量迷迭香酸的迷迭香提取物给药组(1 000、2 000和4 000 mg·kg-1·d-1)对比灌胃.实验6 w后处死动物,测定血清和脑中超氧化物歧化酶(SOD)活力、丙二醛(MDA)含量、谷胱甘肽过氧化物酶(GSH-Px)活性、甘油三酯(TG)水平;以及进行耐缺氧实验. 结果 高剂量的迷迭香酸能明显提高衰老小鼠血清和脑中的SOD活力(P＜0.01)、GSH-Px活性(P＜0.05),降低MDA含量(P＜0.05)和TG水平(P＜0.05),且能明显延长小鼠常压耐缺氧时间(P＜0.05).结论 迷迭香酸通过提高抗氧化酶活性,清除自由基,减少过氧化脂质生成而抗衰老.     

D-半乳糖致衰老模型小鼠皮肤衰老的观察

目的　观察D 半乳糖连续皮下注射对小鼠皮肤衰老指标的影响。　方法　 3月龄KM雌性小鼠 6 0只 ,随机分为空白对照组 (注射生理盐水 )、D 半乳糖低剂量组 (80mg/kg)和D 半乳糖高剂量组 (10 0 0mg/kg) ,连续每日背部皮下注射 4 2天后 ,观察小鼠皮肤中与衰老相关的生化指标及组织病理学变化 ,并用计算机图像分析系统定量分析。　结果　高剂量组小鼠真皮厚度〔(6 2 4 5±4 8 5 ) μm〕较对照组〔(839 3± 5 8 4 ) μm〕显著变薄 (P <0 0 5 ) ,胶原纤维减少 ,排列疏松 ,弹力纤维总面积亦显著减少 (P <0 0 5 ) ;高剂量组皮肤中超氧化物歧化酶 (SOD)活性〔(131 2± 11 5 )U/ml〕、羟脯氨酸含量〔(0 5 7± 0 13) μg/mg〕显著低于对照组的〔(178 1± 2 0 7)U/ml、(0 74± 0 17) μg /mg ,P <0 0 5〕 ,丙二醛 (MDA)含量增加〔(9 4± 1 5 )nmol/ g和 (6 8± 1 5 )nmol/ g ,P <0 0 5〕 ;而低剂量组上述改变则不显著。　结论　每日 10 0 0mg/kgD 半乳糖连续皮下注射 4 2天 ,可导致小鼠皮肤的明显衰老 ,为抗皮肤衰老研究提供一个简便、稳定的实验模型。     

氧化应激对D-半乳糖致衰老小鼠的影响

目的 探讨氧化应激与衰老的相互关系.方法 应用腹腔注射D-半乳糖方法建立衰老模型,观察实验小鼠的外观特征,测定其运动能力、免疫器官指数(胸腺指数、脾脏指数)、脾淋巴细胞增殖活性、血清和脑组织中的SOD及MDA含量,并进行统计学分析.结果 与对照组相比,D-半乳糖腹腔注射后小鼠运动能力、脾指数、胸腺指数及脾细胞增殖活性下降(均P＜0.01),SOD的活性降低(P＜0.01),MDA的含量升高(P＜0.01).结论 氧化应激参与了衰老的发生发展过程,降低了机体的免疫功能和抗氧化能力.     

神经生长液对D-半乳糖衰老模型小鼠的影响

目的　观察中药合剂神经生长液对 D-半乳糖衰老模型小鼠的影响。方法　建立 D-半乳糖致小鼠衰老模型 ,以 Y型电迷宫检测其学习记忆能力 ,分别采用黄嘌呤氧化酶法、TBA比色法测定大脑 SOD活性和 MDA含量 ,观察小鼠服用神经生长液后上述指标的变化。结果　与 D-半乳糖模型组相比 ,神经生长液低、中、高剂量均可显著提高衰老模型小鼠的学习记忆能力 (P<0 .0 5或 P<0 .0 1 ) ,显著增加 D-半乳糖所致衰老模型小鼠大脑 SOD活性、降低 MDA含量 (P<0 .0 5或 P<0 .0 1 )。结论　神经生长液能改善 D-半乳糖所致衰老小鼠的多项体征。     

林蛙油对D-半乳糖致衰老模型小鼠的保护作用

目的 研究林蛙油对D-gal诱导的衰老小鼠大脑学习记忆及抗衰老的作用.方法 小鼠颈背部皮下注射D-gal(120 mg/kg)建立衰老模型,同时以林蛙油(0.05,0.2,0.8g/kg)作为抗衰老实验药物灌服.60 d后.与衰老小鼠比较在行为、记忆上的差异和脑组织内的丙二醛(MDA)含量及超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、Na+-K+-ATPase活性的差异.结果 在水迷宫和跳台实验中林蛙油的学习记忆能力明显长于模型组(P<0.05);D-gal致小鼠的空间认知功能损伤严重,林蛙油组SOD、GSH-Px以及Na+-K+-TPase酶活性高于模型组,而MDA含量低于模型组(P<0.05).结论 林蛙油能显著提高衰老模型小鼠学习记忆能力及脑组织中SOD含量、GSH-Px及其Na+-K+-TPase酶活性,并从分子水平揭示了林蛙油抗衰老的作用机制.     

苦丁茶提取物对D-半乳糖致衰老小鼠学习记忆的影响

目的 观察苦丁茶提取物对D-乳糖致衰老模型小鼠学习记忆能力及抗氧化能力的影响.方法 40只小鼠随机分为正常组,模型组,低剂量组处理组( 10 g/kg),高剂量组处理组(20 g/kg),0.8% D-gal皮下注射6w建立衰老小鼠模型,同时灌胃给予不同剂量苦丁茶提取物.6 w后用Morris水迷宫实验检测小鼠的学习记忆能力,处死小鼠检测脑组织超氧化物歧化酶(SOD)活力以及丙二醛(MDA)含量.结果 与模型组相比,不同剂量处理组小鼠学习记忆成绩均明显提高(P＜0.05),脑组织SOD活力升高(P＜0.05),MDA含量下降(P＜0.05).结论 苦丁茶提取物可改善D-gal衰老模型小鼠的学习记忆能力,其机制可能与其具有抗氧化能力有关.     

枸杞多糖对D-半乳糖诱导衰老小鼠皮肤的影响

目的探讨枸杞多糖的抗皮肤衰老作用。方法健康昆明种雌性小鼠50只,随机分为正常对照组、衰老模型组、枸杞多糖低、中、高剂量组,每组10只。除正常对照组外,其余各组均每日颈背部皮下注射D-半乳糖连续42 d,建立衰老模型。给药组同时给予不同剂量的枸杞多糖。42 d后测定小鼠皮肤组织超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性,脂质过氧化产物丙二醛(MDA)含量,以及皮肤羟脯氨酸(HYP)含量。结果枸杞多糖可增强衰老模型小鼠皮肤组织SOD、GSH-Px活力,降低MDA含量,提高HYP含量。结论枸杞多糖具有延缓D-半乳糖致小鼠皮肤衰老的作用。     

Effect of advanced glycation endproducts on gene expression profiles of human dermal fibroblasts

The Maillard reaction and its end products, AGE-s (Advanced Glycation End products) are rightly considered as one of the important mechanisms of post-translational tissue modifications with aging. We studied the effect of two AGE-products prepared by the glycation of lysozyme and of BSA, on the expression profile of a large number of genes potentially involved in the above mentioned effects of AGE-s. The two AGE-products were added to human skin fibroblasts and gene expression profiles investigated using microarrays. Among the large number of genes monitored the expression of 16 genes was modified by each AGE-preparations, half of them only by both of them. Out of these 16 genes, 12 were more strongly affected, again not all the same for both preparations. Both of them upregulated MMP and serpin-expression and downregulated some of the collagen-chain coding genes, as well as the cadherin- and fibronectin genes. The BSA-AGE preparation downregulated 10 of the 12 genes strongly affected, only the serpin-1 and MMP-9 genes were upregulated. The lysozyme-AGE preparation upregulated selectively the genes coding for acid phosphatase (ACP), integrin chain α5 (ITGA5) and thrombospondin (THBS) which were unaffected by the BSA-AGE preparation. It was shown previously that the lysozyme-AGE strongly increased the rate of proliferation and also cell death, much more than the BSA-AGE preparation. These differences between these two AGE-preparations tested suggest the possibility of different receptor-mediated transmission pathways activated by these two preparations. Most of the gene-expression modifications are in agreement with biological effects of Maillard products, especially interference with normal tissue structure and increased tissue destruction.     

Advanced glycation end-products in senile diabetic and non-diabetic patients with cardiovascular complications

Advanced glycation end products (AGEs) have been reported to contribute to aging and cardiovascular complications. In the present study, the immunoreactivity of AGEs in human serum samples of healthy older subjects (n = 31), senile diabetic patients without cardiovascular complications (n = 33), senile diabetic patients with cardiovascular complications (n = 32), senile non-diabetic patients with cardiovascular complications (n = 30) ,and healthy young subjects (n = 31) were investigated. The patients were selected on clinical grounds from the National Institute of Cardiovascular Disease, Karachi and the Jinnah Postgraduate Medical Centre, Karachi, Pakistan. Fasting blood glucose, HbA_1C and serum fructosamine levels were significantly (P < 0.001) increased in senile diabetic patients with and without cardiovascular complications as compared to non-diabetic senile patients with cardiovascular complications and healthy older subjects. Additionally, serum AGEs were found to be significantly (P < 0.001) increased in senile diabetic patients with cardiovascular complications and senile non-diabetic patients with cardiovascular complications, followed by diabetic patients without cardiovascular complications as compared to healthy older subjects and young control subjects. However, no significant difference was found in the senile diabetic patients without cardiovascular complications and senile non-diabetic patients with cardiovascular complications. In contrast to all four senile groups, serum AGEs were significantly (P < 0.001) lower in young control subjects. The AGEs distribution in the senile groups corroborates the hypothesis that the advanced glycation process might play a role in the development of cardiovascular complications, which are more severe in diabetic patients compared with non-diabetic patients with cardiovascular complications.     

非酶糖化蛋白对体外培养血管内皮细胞生长的影响

目的 观察糖代蛋白终末产物（ＡＧＥｓ）及高糖对体外培养人血管内皮细胞增殖的影响。方法 采用＾３Ｈ标记的腺嘧啶（＾３－ＴｄＲ）掺入法测定细胞增殖情况。结果 在含ＡＧＥｓ的培养液（５μｇ／ｍｌ）中增培养的新生儿脐带静脉内皮细胞,其２小时＾３Ｈ－ＴｄＲ掺入对照组的２．９５倍（Ｐ〈０．０１）,而在含３０ｍｍｏｌ／Ｌ葡萄糖的２液中培养的人胚胎脐带静脉内皮细胞,其２小时＾３Ｈ－ＴｄＲ掺入对照组（１１ｍｍｏｌ／     

吡哆胺和替米沙坦对自发性高血压大鼠心肌重塑相关指标的影响

目的 :观察单用及联用吡哆胺和替米沙坦对自发性高血压大鼠(SHR)心肌重塑相关指标的影响。方法 :48只22周龄雄性SHR,随机分为4组,12只/组。高血压对照组(H组)予以蒸馏水2 ml/d、替米沙坦组(T组)予以替米沙坦6 mg(/kg·d)、吡哆胺组(P组)予以吡哆胺200 mg(/kg·d)、联合干预组(TP组)予以替米沙坦6 mg(/kg·d)+吡哆胺200 mg(/kg·d)灌胃。测定干预前后大鼠尾动脉收缩压。干预16 W后,化学发光法检测血清一氧化氮(NO)、超氧化物歧化酶(SOD),酶联免疫吸附测定法检测血清晚期糖基化终末产物(AGEs),计算左心室重量指数,计算心肌胶原容积分数,实时定量聚合酶链式反应(PCR)检测心肌晚期糖基化终末产物受体(RAGE)信使核糖核酸(mR NA)表达水平。结果 :与H组相比,T组、TP组收缩压下降(P0.01),P组收缩压无明显改变(P0.05);与H组相比,T组、P组、TP组血清NO、SOD浓度升高(P0.01);与T组、P组相比,TP组血清NO浓度、SOD浓度进一步升高(P0.05)。与H组相比,T组、P组、TP组左心室重量指数、心肌胶原容积分数降低(P0.01);与T组、P组相比,TP组左心室重量指数、心肌胶原容积分数进一步降低(P0.05)。与H组(6.71±0.50)mg/L相比,T组(5.99±0.51)mg/L、P组(5.57±0.91)mg/L、TP组(5.24±0.63)mg/L血清AGEs降低(P0.01);与T组、P组相比,TP组血清AGEs进一步降低(P0.05)。吡哆胺和替米沙坦两因素在左心室重量指数、心肌胶原容积分数中存在协同效应(P0.05)。与H组(0.053±0.010)相比,T组(0.035±0.010)、P组(0.036±0.005)、TP组(0.024±0.007)RAGE mR NA表达水平降低(P0.01);与T组、P组相比,TP组RAGE mR NA表达水平进一步降低(P0.05)。差异均有统计学意义。结论:吡哆胺有独立于降压因素之外的改善SHR体内氧化应激状态、改善心肌重塑相关指标的作用,联用吡哆胺和替米沙坦可以从不同途径打破氧化应激介导的AGEs-RAGE和肾素—血管紧张素系统的协同作用,进一步改善心肌重塑相关指标。     

Advanced glycation end products in senile diabetic and nondiabetic patients with cataract

BackgroundAdvanced glycation end products (AGE) have been reported to contribute to aging and cataract formation in the lens. In the present study, AGE immunoreactivity in human serum samples of normal senile subjects (n=31), senile diabetic patients without cataract (n=33), senile diabetic patients with cataract (n=30), senile nondiabetic with cataract (n=30), and normal young subjects (n=31) was investigated.MethodsA noncompetitive ELISA with polyclonal anti-AGE antibody was performed. The patients were selected on clinical grounds from Eye Ward, Jinnah Postgraduate Medical Centre, Karachi, Pakistan.ResultsFasting blood glucose, glycosylated hemoglobin, and serum fructosamine were estimated. Fasting blood glucose, HbA_1C, and serum fructosamine levels were significantly (P<.001) increased in senile diabetic patients with and without cataract as compared to nondiabetic senile patients with cataract and senile control subjects. However, the serum AGEs were found to be significantly (P<.001) increased in senile diabetic patients with cataract and senile nondiabetic patients with cataract followed by the diabetic patients without cataract as compared to senile control and young control subjects. In contrast to all four senile groups, the serum AGEs were significantly (P<.001) lower in young control subjects.ConclusionsThe AGE distribution in the senile groups corroborates the hypothesis that the advanced glycation process might have a role in cataract formation, which in diabetic patients occurs vigorously as compared with nondiabetic cataract patients.     

糖基化终产物对正常人血小板内皮型一氧化氮合酶活性及其表达的影响

目的 通过观察糖基化终产物（AGEs)对血小板内皮型一氧化氮合酶（eNOS)活性及其表达的影响，探讨AGEs损伤血小板的机制。方法 健康成人6例，取外周静脉血，用凝胶色谱柱法收集纯化的血小板悬液，分别与三种浓度的AGEs(100μg/ml,200μg/ml，400μg/ml）孵充300min，部分血小板用位素二步色谱法测定血小板一氧化氮合酶（NOS）活性；部分血小板用免疫沉淀法制备经eNOS蛋白，蛋白印迹法检测eNOS蛋白表达水平，结果 AGEs明显抑制正常人血小板NOS纯化eNOS蛋白，蛋白印迹法检测eNOS蛋白表达水平。结果 AGEs明显抑制正常人血小板NOS活性且呈浓度依赖性。正常人血小板表达eNOS；AGEs干预30min不影响eNOS表达水平。结论 AGEs通过抑制血小板eNOS活性而激活血小板，这种作用不是通过降低血小板eNOS表达实现的。     

肝纤维化大鼠晚期糖基化终末产物变化及氨基胍的干预作用

目的研究四氯化碳诱导的实验性大鼠肝纤维化不同阶段体内晚期糖基化终末产物的变化情况，以及晚期糖基化终末产物抑制剂氨基胍对肝纤维化的干预作用。方法sD大鼠分为3组：对照组、模型组、氨基胍干预组。检测四氯化碳诱导的大鼠肝纤维化及氨基胍干预后不同阶段血清及肝组织匀浆晚期糖基化终末产物含量。血清透明质酸水平，并对照观察肝脏的组织病理学改变进行肝纤维化程度评分。结果血清及肝组织匀浆晚期糖基化终末产物含量、透明质酸水平以及肝纤维化评分，肝纤维化l2周组高于对照组（t值分别为3．26、3．49、4．70和25．4，P值均〈0．01），氨基胍干预组低于肝纤维化组（t值分别为2．11，2．11、2．93和2．47，P值均〈0．05）。血清及肝组织匀浆晚期糖基化终末产物含量与血清透明质酸水平呈线性相关。结论肝纤维化晚期血清及肝组织匀浆晚期糖基化终末产物含量是增高的。一定程度上，体内晚期糖基化终末产物水平可以反映肝纤维化情况。氨基胍对四氯化碳诱导的实验性大鼠肝纤维化有一定的干预作用。     

晚期糖基化终末产物对高血压及其合并早期肾损害相关性分析

目的:通过测定高血压病(EH)患者与正常人血清晚期糖基化终末产物(AGEs)及尿微量白蛋白(mALB)的水平,探讨AGEs与EH及其早期肾功能损害的关系,为高血压发病机制及其早期肾功能损害防治研究提供理论依据。方法:筛选EH患者100例,其中EH mALB正常组42例,EH mALB异常组58例。所有入选者分别用放免法、ELISA法测晨尿微量白蛋白(mALB)、AGEs水平。结果:与对照组比较,EH mALB正常组、EH mALB异常组血清AGEs显著增高(P<0.001);与EHmALB正常组比较,EHmALB异常组血清AGEs显著增高(P<0.001)。相关分析AGEs与SBP、PP、mALB之间有显著的正相关性(r=0.448-0.687,P<0.001);mALB与SBP、PP之间有显著的正相关性(r=0.271-0.354,P<0.001)。结论:AGEs、SBP共同参与高血压病肾脏功能的损害,SBP随AGEs水平升高而增高。