Inhibition of advanced glycation endproducts formation by Korean thistle,Cirsium maackii

Objective:To evaluate inhibitoy potential of seven Korean thistles against the advanced glyeatinn endproducts(AGE) formation as well as to identify responsible compounds from the most active species.Methods:We used an in vitro AGE inhibition assay to evaluate the antidiabetic complication potential of the methanol extracts of the selected Korean thistles.Results:Among the seven Korean thistles,the leaves of Cirsium maackii(C.maackii) exhibited the most significant inhibitory activity against AGE formation.By means of bioassay-directed fractionation,a lignan.chlorogenic acid and 14 flavonoids were isolated from the active ethyl acetate soluble fraction of a methanol extract from C.maackii leaves.Luteolin and its 5-O-glueoside have been previously isolated:however,a lignan and 13 known compounds were isolated for the first lime from C maackii leaves in this study.Most of the isolated compounds exhibited inhibitory activities against potential AGE formation.Among them,cernuoside was shown to be the most potent AGE inhibitor with an IC_(50) value of 21.21 μ mol/L.Most importantly,two major flavonoids,luteolin and ils 5-O-glucoside,also significantly inhibited AGE formation,with IC_(50) values of36.33 and 37.47 μmol/L,respectively.Structure activity relationship revealed that the presence of free 3'and 4' dihydroxyl group in flavonoids skeleton played an important role in AGE inhibition.Conclusions:These results indicate that C.maackii and C maackii-derived flavonoids might be explored further to develop therapeutic agents for the prevention of diabetic eoniplicalions due to their significant inhibitory activity against AGE formation.

Influence of tonifying kidney recipe on advanced glycation endproducts and lipid peroxidation in ovariectomized rats

BACKGROUND： Previous studies have demonstrated that reduced estrogen levels may accelerate the formation of advanced glycation endproducts （AGE） in brain tissue, raise the concentration of lipid peroxidation products in vivo, and speed up deterioration of learning and memory. A tonifying kidney recipe is hypothesized to improve the ability of learning and memory in ovariectomized rats by downregulating AGE and lipid peroxidation products. OBJECTIVE： To simulate a postmenopausal state, bilateral ovariectomy （OVX） was performed in rats, and the effects of tonifying kidney recipe （TKR） on AGE and lipid peroxidation in the rat cerebral cortex, hippocampus, and blood serum levels was measured. In addition, the effects on learning and memory were evaluated, and the effect of AGE -specific inhibitor aminoguanidine （AG） was compared with TKR. DESIGN, TIME AND SETTING： A randomized, in vivo, control experiment was performed at the scientific research center （Provincial Key Laboratory） in the Fourth Hospital of Hebei Medical University （Shijiazhuang, Hebei Province, China） from May 2005 to January 2007. MATERIALS： Forty healthy, adult, female, Sprague Dawley rats were used for this study. TKR was composed of prepared rehmannia rhizome, epimedium herb, desert-living cistanche, and Szechwan lovage rhizome, which were provided by Shijiazhuang Medical Materials Company （China）. A TKR extraction was prepared for further use. AG was provided by Sigma （USA）. Forty rats were randomly divided into four groups： sham, OVX, AG, and TKR, with 10 rats in each group. METHODS： The rat ovaries were resected in the OVX, AG, and TKR groups, whereas the same volume of fat was resected in the sham group. At four weeks after OVX, the AG group received 1% AG water solution by lavage; the TKR group was administrated by lavage once per day at a dose of 6.3 g （crude drug）/kg; OVX and sham groups received equal volumes of tap water. MAIN OUTCOME MEASURES： Learning and memory behavior of rats was tested in a Y-electric maze 16 weeks after the OVX procedure. The contents of advanced glycation endproducts in the rat cerebral cortex, hippocampus, serum, and urine were detected by competitive ELISA and spectrofluorophotometer. The contents of lipid peroxidation in rat serum, cerebral cortex, and hippocampus were assayed using a biochemical method. RESULTS： Compared with the sham group, serum content of advanced glycation endproducts in the OVX group was significantly increased, and lipid peroxidation content increased in cerebral cortex, hippocampus, and serum （t = 3.04-4.22, P 〈 0.05 0.01）. Both AG and TKR decreased the amount of AGE in cerebral cortex and serum （t = 2.53, 3.64, P 〈 0.05, 0.01）, increased AGE urine content （t = 3.25 4.87, P 〈 0.01）, and decreased lipid peroxidation content in cerebral cortex, hippocampus, and serum （t = 2.80 3.70, P 〈 0.05 0.01）. In comparison to the OVX and sham groups, the correct escape rate in the Y-electric maze was significantly increased （t = 3.46, 3.28, P 〈 0.01）, and escape latency was significantly decreased （t=3.12, 2.48 P 〈 0.05） in the AG and TKR groups, which indicated that both AG and TKR improved learning and memory The OVX group had a significantly lower correct escape compared with the sham group （t = 4.21, P 〈 0.01 ）.CONCLUSION： The tonifying kidney recipe decreased deposition of advanced glycation endproducts and lipid peroxidation in ovariectomized rats, and concomitantly improved learning and memory. The effect of TKR was equal to that of AG.

Adherence to Mediterranean diet and advanced glycation endproducts in patients with diabetes

BACKGROUND In recent years,American Diabetes Association started to strongly advocate the Mediterranean diet(MD)over other diets in patients with diabetes mellitus(DM)because of its beneficial effects on glycemic control and cardiovascular(CV)risk factors.Tissue levels of advanced glycation endproducts(AGEs)emerged as an indicator of CV risk in DM.Skin biopsy being invasive,the use of AGE Reader has been shown to reflect tissue AGEs reliably.AIM To examine the association between adherence to MD and AGEs in patients with DM type II.METHODS This cross-sectional study was conducted on 273 patients with DM type II.A survey questionnaire was composed of 3 separate sections.The first part of the questionnaire included general data and the habits of the participants.The second part aimed to assess the basic parameters of participants’diseases and associated conditions.The third part of the questionnaire was the Croatian version of the 14-item MD service score(MDSS).AGEs levels and associated CV risk were measured using AGE Reader(DiagnOptics Technologies BV,Groningen,The Netherlands).RESULTS A total of 27(9.9%)patients fulfilled criteria for adherence to MD,with a median score of 8.0(6.0-10.0).Patients with none/limited CV risk had significantly higher percentage of MD adherence in comparison to patients with increased/definite CV risk(15.2%vs 6.9%,P=0.028),as well as better adherence to guidelines for nuts(23.2%vs 12.6%,P=0.023)and legumes(40.4%vs 25.9%,P=0.013)consumption.Higher number of patients with glycated hemoglobin(HbA1c)<7%adhered to MD when compared to patients with HbA1c>7%(14.9%vs 7.3%,P=0.045).Moreover,those patients followed the MDSS guidelines for eggs(33.0%vs 46.8%,P=0.025)and wine(15.6%vs 29.8%,P=0.006)consumption more frequently.MDSS score had significant positive correlation with disease duration(r=0.179,P=0.003)and negative correlation with body mass index(BMI)values(r=-0.159,P=0.008).In the multiple linear regression model,BMI(β±SE,-0.09±0.04,P=0.037)and disease duration(β±SE,0.07±0.02,P<0.001)remained significant independent correlates of the MDSS score.Patients with HbA1c>7%think that educational programs on nutrition would be useful for patients in significantly more cases than patients with HbA1c<7%(98.9%vs 92.6%,P=0.009).CONCLUSION Although adherence to MD was very low among people with diabetes,we demonstrated that adherence to MD is greater in patients with lower CV risk,longer disease duration,and well-controlled glycaemia.

Fluorescent advanced glycation end-products (ages) detected by spectro-photofluorimetry, as a screening tool to detect diabetic microvascular complications

BACKGROUND: Advanced glycation end-products (AGEs) are one of the mechanisms related to diabetic vascular complications. However, since AGEs are multiple and heterogeneous moieties, there is no universally accepted method to measure them for clinical purposes. The aim of this work was to study the utility of a simple fluorimetric assay as predictor of complications. METHODS: Blood samples from 102 type 2 diabetic patients were obtained to assess glucose, glycosylated haemoglobin, creatinine, lipoproteins and C Reactive Protein (CRP), fluorescent AGES by spectrophotofluorimetry and non-fluorescent AGEs by measurement of N(ε)-carboxymethyl-Lysine (CML) using an ELISA kit in a subsample of 82 patients. Urinary fluorescent AGEs, albumin and creatinine were also measured in a morning urine sample. Microvascular complications were studied by ophthalmologic examination, albuminuria and peripheral nerve conduction velocity. RESULTS: Patients without microvascular complications had significantly lower levels of both serum and urinary AGEs. CML was associated with retinopathy. Multiple regression analysis confirmed that AGEs, length of diabetes and glycosylated haemoglobin were all variables associated with diabetic complications, in this sample. CONCLUSIONS: A simple fluorimetric assay to measure low molecular weight fluorescent AGEs, and CML could be employed as screening tools to predict diabetic complications, at a primary care setting. AGEs should probably be considered as another therapeutic target in diabetes management.

Advanced Glycation Endproducts in 35 Types of Seafood Products Consumed in Eastern China

Advanced glycation endproducts(AGEs) have been recognized as hazards in processed foods that can induce chronic diseases such as cardiovascular disease, diabetes, and diabetic nephropathy. In this study, we investigated the AGEs contents of 35 types of industrial seafood products that are consumed frequently in eastern China. Total fluorescent AGEs level and Nε-carboxymethyl-lysine(CML) content were evaluated by fluorescence spectrophotometry and gas chromatography-mass spectrometry(GC-MS), respectively. The level of total fluorescent AGEs in seafood samples ranged from 39.37 to 1178.3 AU, and was higher in canned and packaged instant aquatic products that were processed at high temperatures. The CML content in seafood samples ranged from 44.8 to 439.1 mg per kg dried sample, and was higher in roasted seafood samples. The total fluorescent AGEs and CML content increased when seafood underwent high-temperature processing, but did not show an obvious correlation. The present study suggested that commonly consumed seafood contains different levels of AGEs, and the seafood processed at high temperatures always displays a high level of either AGEs or CML.

Role of the receptor for advanced glycation end products in hepatic fibrosis

AIM： To study the role of advanced glycation end products （AGE） and their specific receptor （RAGE） in the pathogenesis of liver fibrogenesis. METHODS： In vitro RAGE expression and extracellular matrix-related gene expression in both rat and human hepatic stellate cells （HSC） were measured after stimulation with the two RAGE ligands, advanced glycation end product-bovine serum albumin （AGE- BSA） and N＇-（carboxymethyl） lysine （CML）-BSA, or with tumor necrosis factor-α （TNF-α）. In vivo RAGE expression was examined in models of hepatic fibrosis induced by bile duct ligation or thioacetamide. The effects of AGE-BSA and CML-BSA on HSC proliferation, signal transduction and profibrogenic gene expression were studied in vitro. RESULTS： In hepatic fibrosis, RAGE expression was enhanced in activated HSC, and also in endothelial cells, inflammatory cells and activated bile duct epithelia. HSC expressed RAGE which was upregulated after stimulation with AGE-BSA, CML-BSA, and TNF-α.RAGE stimulation with AGE-BSA and CML-BSA did not alter HSC proliferation, apoptosis, fibrogenic signal transduction and fibrosis- or fibrolysis-related gene expression, except for marginal upregulation of procollagen α1（ I ） mRNA by AGE-BSA. CONCLUSION： Despite upregulation of RAGE in activated HSC, RAGE stimulation by AGE does not alter their fibrogenic activation. Therefore, RAGE does not contribute directly to hepatic fibrogenesis.

Injury of cortical neurons is caused by the advanced glycation end products-mediated pathway

Advanced glycation end products lead to cell apoptosis, and cause cell death by increasing endoplasmic reticulum stress. Advanced glycation end products alone may also directly cause damage to tissues and cells, but the precise mechanism remains unknown. This study used primary cultures of rat cerebral cortex neurons, and treated cells with different concentrations of glycation end products （50, 100, 200, 400 mg/L）, and with an antibody for the receptor of advanced glycation end products before and after treatment with advanced glycation end products. The results showed that with increasing concentrations of glycation end products, free radical content increased in neurons, and the number of apoptotic cells increased in a dose-dependent manner. Before and after treatment of advanced glycation end products, the addition of the antibody against advanced glycation end-products markedly reduced hydroxyl free radicals, malondialdehyde levels, and inhibited cell apoptosis. This result indicated that the antibody for receptor of advanced glycation end-products in neurons from the rat cerebral cortex can reduce glycation end product-induced oxidative stress damage by suppressing glycation end product receptors. Overall, our study confirms that the advanced glycation end products-advanced glycation end products receptor pathway may be the main signaling pathway leading to neuronal damage.

Circulating levels of advanced glycation end products in females with polycystic ovary syndrome:a meta-analysis

Polycystic ovary syndrome(PCOS)is a common endocrine disorder characterized by a hormonal imbalance that affects females of reproductive age.The association between advanced glycation end products(AGEs)and PCOS has attracted considerable attention in recent years,highlighting the potential of AGEs as biomarkers for this disorder.In the present systematic review and meta-analysis,we aimed to examine the association between AGEs and PCOS,evaluate their potential as biomarkers,and improve our understanding of the pathophysiology of PCOS and its associated metabolic complications.A literature search was performed using various databases from January 2000 to March 2023 to identify relevant studies investigating the association between AGEs and PCOS.Pooled effect estimates were calculated using standardized mean differences(SMD)with 95%confidence intervals(CIs).Sub-group and meta-regression analyses were performed to examine potential sources of heterogeneity.The meta-analysis included six studies with a total of 623 participants.Our results revealed a significant increase in circulating AGE levels in females with PCOS compared to healthy females(SMD=2.35;95%CI:1.10-3.60;P<0.001).Significant heterogeneity was observed between the studies(I2=96.37%;P<0.001),indicating the presence of several factors influencing the association.Sub-group analyses based on body mass index,age,and homeostatic model assessment for insulin resistance indicated differential effects of AGEs on specific sub-groups.This systematic review and meta-analysis support the association between elevated AGE levels and PCOS,thereby suggesting the potential role of AGEs as biomarkers in PCOS.

Intrafollicular Soluble RAGE Benefits Embryo Development and Predicts Clinical Pregnancy in Infertile Patients of Advanced Maternal Age Undergoing In Vitro Fertilization

Soluble receptor for advanced glycation end products（s RAGE） can decoy the toxic AGEs and is considered to be a protective factor.This study aimed to evaluate the correlation between intrafollicular s RAGE levels and clinical outcomes in infertile women of young or advanced maternal age（AMA） undergoing in vitro fertilization（IVF）.A total of 62 young women and 62 AMA women who would undergo IVF were included in this prospective study.The intrafollicular s RAGE concentration was measured to determine its association with the number of retrieved oocytes,fertilized oocytes,high-quality embryos or achievement of clinical pregnancy in young and AMA women,respectively.Besides,correlations between sR AGE and age or follicle-stimulating hormone（FSH） were examined.We found that the intrafollicular s RAGE levels were higher in young patients than those in AMA patients,suggesting that the s RAGE levels were inversely correlated with age.In young patients,sR AGE showed no correlation with the number of retrieved oocytes,fertilized oocytes,high-quality embryos or achievement of clinical pregnancy.But it was found that AMA patients with more retrieved oocytes,fertilized oocytes and high-quality embryos demonstrated higher sR AGE levels,which were a prognostic factor for getting clinical pregnancy independent of age or FSH level.In conclusion,the s RAGE levels decrease with age.Elevated intrafollicular s RAGE levels indicate good follicular growth,fertilization and embryonic development,and successful clinical pregnancy in AMA women,while in young women,the role of s RAGE may not be so predominant.

血清DKK1、sRAGE、hCAP-18对急性淋巴细胞白血病患儿的诊断和预后不良的预测价值

目的 探讨血清Dickkopf相关蛋白1(DKK1)、可溶性晚期糖基化终末产物受体(sRAGE)、人类阳离子抗菌蛋白-18(hCAP-18)对急性淋巴细胞白血病患儿的诊断和预后不良的预测价值。方法 选取2020年4月至2022年4月南京鼓楼医院集团宿迁医院收治的84例急性淋巴细胞白血病患儿作为研究组,包括标危28例、中危36例、高危20例。另选取同期在南京鼓楼医院集团宿迁医院体检的84例健康儿童作为对照组。收集所有研究对象的临床资料。采用酶联免疫吸附试验检测所有研究对象血清DKK1、sRAGE、hCAP-18水平。对研究组患儿进行为期1年的随访,并按随访结果分为预后不良组和预后良好组。绘制受试者工作特征(ROC)曲线分析血清DKK1、sRAGE、hCAP-18在急性淋巴细胞白血病患儿诊断和预后不良的预测价值。采用多因素Logistic回归分析影响急性淋巴细胞白血病发生的因素。结果 与对照组相比,研究组患儿白细胞计数(WBC)、DKK1水平显著升高,sRAGE、hCAP-18水平显著降低,差异均有统计学意义(P<0.05)。与研究组标危患儿相比,中危和高危患儿血清DKK1水平显著升高,sRAGE、hCAP-18水平显著降低,差异均有统计学意义(P<0.05);与中危患儿相比,高危患儿血清DKK1水平显著升高,sRAGE、hCAP-18水平显著降低,差异均有统计学意义(P<0.05)。血清DKK1、sRAGE、hCAP-18联合诊断急性淋巴细胞白血病的曲线下面积(AUC)显著高于血清DKK1、sRAGE、hCAP-18单独诊断的AUC(Z=2.820,P=0.005;Z=5.170,P<0.001;Z=5.272,P<0.001)。多因素Logistic回归分析结果显示,血清WBC、DKK1、sRAGE、hCAP-18是急性淋巴细胞白血病发生的影响因素(P<0.05)。随访1年后,26例患儿纳入预后不良组,58例患儿纳入预后良好组。与预后良好组相比,预后不良组患儿血清DKK1水平显著升高,sRAGE、hCAP-18水平显著降低,差异均有统计学意义(P<0.05)。与预后不良组中危患儿相比,高危患儿DKK1水平显著升高,sRAGE、hCAP-18水平显著降低,差异均有统计学意义(P<0.05)。血清DKK1、sRAGE、hCAP-18联合预测急性淋巴细胞白血病患儿预后不良的AUC明显高于血清DKK1、sRAGE、hCAP-18单独预测的AUC(Z=2.312,P=0.021;Z=3.160,P=0.002;Z=2.926,P=0.003)。结论 急性淋巴细胞白血病患儿血清DKK1高表达,sRAGE、hCAP-18低表达,且三者联合检测对急性淋巴细胞白血病具有一定的诊断和预后预测价值。

新生儿呼吸窘迫综合征患儿血清HMGB1、RAGE、BMP-7表达及与肺部超声评分的相关性

目的 分析新生儿呼吸窘迫综合征(NRDS)患儿血清高迁移率族蛋白1(HMGB1)、晚期糖基化终末产物受体(RAGE)、骨形态发生蛋白-7(BMP-7)表达及与肺部超声评分的相关性。方法 选择2018年6月至2023年6月收治的100例NRDS患儿纳入观察组,同期选择100例健康新生儿纳入对照组,均进行血清HMGB1、RAGE、BMP-7检测和肺部超声检查及评分。根据NRDS患儿肺部超声评分将其分为低评分组(评分<12分)、中评分组(评分12~24分)及高评分组(评分>24分)。比较两组研究对象的血清HMGB1、RAGE、BMP-7水平和肺部超声评分;比较不同肺部超声评分患儿的血清HMGB1、RAGE、BMP-7水平;分析观察组中血清HMGB1、RAGE、BMP-7表达与肺部超声评分的相关性。结果 观察组的血清HMGB1、RAGE及BMP-7水平及肺部超声评分高于对照组(P<0.05)。肺部超声显示低评分组47例,中评分组33例,高评分组20例;低评分组的血清HMGB1、RAGE及BMP-7水平低于中评分组和高评分组(P<0.05);中评分组的血清HMGB1、RAGE及BMP-7水平低于高评分组(P<0.05)。Spearman相关性分析结果显示,血清HMGB1、RAGE、BMP-7表达与肺部超声评分呈正相关(P<0.05)。结论 NRDS患儿血清HMGB1、RAGE、BMP-7表达及肺部超声评分均会明显增高,血清HMGB1、RAGE、BMP-7表达与肺部超声评分呈正相关。

绞股蓝皂苷对AGEs诱导下人肾小球系膜细胞中RAGE及转化生长因子-β_1表达的影响

目的观察绞股蓝皂苷(gypenosides,GP)对晚期糖基化终末产物(advanced glycation endproducts,AGEs)诱导下人肾小球系膜细胞(human mesangial cells,HMCs)中晚期糖基化终末产物受体(RAGE)及转化生长因子-β_1(TGF-β1)表达的影响。方法将体外培养的(体积分数为0.15的FBS的DMEM低糖培养液)人肾小球系膜细胞(HMCs)分为4组:正常组、模型组、GP组、阳性对照组。除正常组外,其余组均再给予200 mg·L^(-1)AGEs刺激。此外,GP组中加入不同浓度GP(25、75、175 mg·L^(-1))进行干预,阳性对照组中加入氨基胍盐酸盐(10^(-1)mmol·L^(-1))。应用Western blot技术检测各实验组中RAGE和TGF-β_1蛋白的表达;RT-PCR技术检测各实验组中RAGE和TGF-β_1 mRNA的表达。结果模型组AGEs诱导下HMCs中RAGE、TGF-β1蛋白及mRNA表达水平明显高于正常组(P<0.01);与模型组比较,GP组中GP可明显下调HMCs中RAGE、TGF-β_1蛋白及mRNA的表达,且呈浓度依赖性(P<0.01)。结论 GP可降低AGEs诱导下HMCs中RAGE的表达,阻断AGEs-RAGE信号通路,并下调下游因子TGF-β_1的表达,进而延缓糖尿病肾病纤维化进程。

AGEs-RAGE通路阻断对血管平滑肌细胞迁移能力的影响

研究AGEs-RAGE通路阻断对糖尿病大鼠主动脉血管平滑肌细胞(VSMCs)迁移能力的影响并探讨其可能机制。以晚期糖基化终产物受体(RAGE)抗体预孵VSMCs后,再用晚期糖基化终产物(AGEs)刺激VSMCs细胞,Transwell法检测VSMCs迁移能力变化,MTT法及ELISA分别检测VSMCs增殖及p27蛋白表达的变化,DCFH法测定VSMCs细胞内活性氧(ROS)水平,RT-PCR法测定NOX1 mRNA表达水平。结果显示:RAGE抗体可以明显抑制AGEs诱导的VSMCs迁移作用(P<0.01),且抑制迁移作用较其抑制增殖作用强,细胞ROS水平降低(P<0.01),NOX1 mRNA表达量下降但P27蛋白表达变化不大。实验结果表明,用RAGE抗体阻断AGEs-RAGE通路可以抑制VSMCs的迁移,其机制可能与其下调NOX1mRNA表达从而降低细胞内氧化应激水平有关。

AGEs通过RAGE促进Jurkat细胞分泌肿瘤坏死因子-α

目的:探讨晚期糖化终产物(AGEs)与晚期糖化终产物受体(RAGE)结合对Jurkat细胞分泌肿瘤坏死因子-α(TNF-α)的影响及相关信号分子的变化。方法:不同浓度AGEs作用于植物血球凝集素(PHA)预刺激的Jurkat细胞24小时,MTT检测AGEs对细胞生存率的影响,ELISA检测AGEs诱导Jurkat细胞分泌TNF-α;Western blot检测AGEs诱导Jurkat细胞表达RAGE和p38MAPK、JNK磷酸化水平。结果:AGEs作用于PHA预刺激Jurkat细胞24小时对细胞生存率无明显影响;AGEs促进Jurkat细胞表达RAGE,增加炎症因子TNF-α分泌,抗RAGE抗体明显抑制AGEs诱导的TNF-α分泌。AGEs可引起p38MAPK、JNK磷酸化,p38MAPK、JNK的抑制剂明显抑制AGEs诱导的TNF-α表达。结论:AGEs通过RAGE促进PHA预刺激的Jurkat细胞分泌炎症因子TNF-α,其机制与激活p38MAPK、JNK途径有关。

冠心病合并2型糖尿病患者的血清AGEs与内皮依赖性血管舒张功能关系探讨

目的 探讨冠心病合并 2型糖尿病 (2 - DM)患者内皮依赖性血管舒张功能与血清糖基化终产物(AGEs)水平变化及二者的关系。方法 选择 6 0例冠心病合并 2 - DM患者为治疗组 ,在冠心病综合治疗的基础上给予胰岛素或其他降糖药物控制血糖 ;另选 6 0例查体健康者为对照组。检测两组血清 AGEs、NO水平 ,采用 Jud-kins法进行选择性冠状动脉造影 ,高分辨超声技术检测肱动脉的血管舒张功能。结果 治疗组治疗前后血清AGEs、NO水平及内皮依赖性血管舒张功能均与对照组有明显差异 (P <0 .0 5 ,<0 .0 1) ,且治疗组治疗前后上述指标亦均有明显差异 (P <0 .0 1)。内皮依赖性血管舒张功能减弱程度与血清 AGEs水平呈负相关 (r =- 0 .39,P <0 .0 1) ,血清 NO水平与血清 AGEs水平呈负相关 (r=- 0 .31,P <0 .0 1)。结论 冠心病合并 2 - DM患者内皮依赖性血管舒张功能明显减弱 ,血清 AGEs水平明显升高 ,可能是 AGEs削弱了其内皮依赖性血管舒张功能。长期良好的血糖控制可降低冠心病合并 2 - DM患者血清 AGEs水平 ,增加血液中 NO水平 。

p38MAPK/eNOS信号通道在胰高血糖素样肽-1抑制AGEs诱导的人脐静脉内皮细胞凋亡中的作用

目的探讨p38MAPK信号通道在胰高血糖素样肽-1(GLP-1)拮抗糖基化终末产物(AGEs)诱导的人脐静脉内皮细胞凋亡的作用。方法实验组分为对照组、AGEs组、GLP-1组、AGEs+GLP-1组、AGEs+抑制剂组及AGEs+GLP-1+抑制剂组,western blot技术检测p-p38MAPK/p38MAPK、p-e NOS/e NOS蛋白表达情况,Annexin V/PI流式检测细胞凋亡率。结果与对照相比较,单独加入AGEs或GLP-1可分别导致p-p38MAPK蛋白表达水平明显上升(P=0.001)或下降(P<0.001);与对照组相比AGEs可显著降低p-e NOS表达水平(P=0.007),而予以GLP-1或p38MAPK抑制剂(SB203580)预处理后,受抑制的e NOS蛋白表达水平再次显著升高(P=0.004);在AGEs组加入SB203580或GLP-1预处理后,AGEs诱导的细胞凋亡率均显著下降(P<0.001,P<0.001)。结论 GLP-1至少部分通过抑制p38MAPK蛋白磷酸化,上调磷酸化e NOS蛋白的表达,对人脐静脉内皮细胞起到抗凋亡的保护作用。

AGEs对老年大鼠内皮细胞NF-κB活性与Fn mRNA表达的影响

目的:探讨糖基化终末产物(AGEs)对老年大鼠内皮细胞中NF-κB活性及纤维结合蛋白fibronectin(Fn)mRNA表达的影响。方法:取24月龄的SD大鼠主动脉内皮细胞进行原代培养,实验分为3组。A组葡萄糖(5mmol/L)的对照组;B组AGEs(25mg/L,48h)处理组;C组AGEs(50mg/L,48h)处理组。采用荧光免疫化学法检测NF-κB活性和逆转录聚合酶链式反应(RT-PCR)检测FnmRNA的表达。结果:与对照组相比,AGEs呈浓度依赖性地诱导NF-κB的激活(P<0.05)和上调FnmRNA表达(P<0.05)。结论:AGEs呈浓度依赖性地诱导内皮细胞NF-κB的活化、Fn的基因表达上调,这可能与糖尿病慢性血管并发症的发生发展有关。

Pitavastatin attenuates AGEs-induced mitophagy via inhibition of ROS generation in the mitochondria of cardiomyocytes

This study aimed to investigate whether pitavastatin protected against injury induced by advanced glycation end products products（AGEs） in neonatal rat cardiomyocytes,and to examine the underlying mechanisms.Cardiomyocytes of neonatal rats were incubated for 48 hours with AGEs（100 μg/mL）,receptor for advanced glycation end products（RAGE）,antibody（1 μg/mL） and pitavastatin（600 ng/mL）.The levels of p62 and beclinl were determined by Western blotting.Mitochondrial membrane potential（△Ψm） and the generation of reactive oxygen species（ROS） were measured through the JC-1 and DCFH-DA.In the AGEs group,the expression of beclinl was remarkably increased compared to the control group,while the expression of p62 was significantly decreased.AGEs also markedly decreased △Ψm and significantly increased ROS compared with the control group.After treatment with RAGE antibody or pitavastatin,the level of beclinl was markedly decreased compared with the AGEs group,but the level of p62 was remarkably increased.In the AGEs ＋ RAGE antibody group and AGEs＋ pitavastatin group,△Ψm was significantly increased and ROS was remarkably decreased compared with the AGEs group.In conclusion,AGEs-RAGE may induce autophagy of cardiomyocytes by generation of ROS and pitavastatin could protect against AGEs-induced injury against cardiomyocytes.

Food-advanced glycation end products aggravate the diabetic vascular complications via modulating the AGEs/RAGE pathway

The aim of this study was to investigate the effects of high-advanced glycation end products （AGEs） diet on diabetic vascular complications. The Streptozocin （STZ）-induced diabetic mice were fed with high-AGEs diet. Diabetic characteristics, indicators 3f renal and cardiovascular functions, and pathohistology of pancreas, heart and renal were evaluated. AGEs/RAGE/ROS pathway parameters were determined. During the experiments, the diabetic mice exhibited typical characteristics including weight loss, polydipsia, polyphagia, polyuria, high-blood glucose, and low-serum insulin levels. However, high-AGEs diet effectively aggravated these diabetic sharacteristics. It also increased the 24-h urine protein levels, serum levels of urea nitrogen, creatinine, c-reactive protein （CRP）, low density lipoprotein （LDL）, tumor necrosis factor-a （TNF-a）, and interleukin-6 （IL-6） in the diabetic mice. High-AGEs diet deteriorated the histology of pancreas, heart, and kidneys, and caused structural alterations of endothelial ceils, mesangial cells and podocytes in renal ：ortex. Eventually, high-AGEs diet contributed to the high-AGE levels in serum and kidneys, high-levels of reactive oxygen species ＇,ROS） and low-levels of superoxide dismutase （SOD） in serum, heart, and kidneys. It also upregulated RAGE mRNA and protein expression in heart and kidneys. Our results showed that high-AGEs diet deteriorated vascular complications in the diabetic mice. The activation of AGEs/RAGE/ROS pathway may be involved in the pathogenesis of vascular complications in diabetes.

血清sRAGE水平、EOS计数和FEV1检测评估支气管哮喘病情严重程度及结局转归的临床价值

目的探讨血清可溶性晚期糖基化终末产物受体(sRAGE)水平、嗜酸性粒细胞(EOS)计数和第1秒用力呼气容积(FEV1)评估支气管哮喘病情严重程度及结局转归的临床价值。方法回顾性选取2019年7月至2021年10月长江大学附属仙桃市第一人民医院收诊的139例支气管哮喘患者为研究组,另选取同期135名体检健康者为对照组。根据研究组患者病情严重程度分为轻中度组(n=102)、重度组(n=37);另根据研究组患者结局转归情况分为转归良好组(n=123)和转归不良组(n=16)。采用酶联免疫吸附试验检测血清sRAGE水平;采用全血细胞计数仪进行EOS计数;肺功能仪检测所有受试者FEV1;采用受试者工作特征(ROC)曲线评估血清sRAGE水平、EOS计数和FEV1检测对支气管哮喘病情严重程度及结局转归的诊断价值。结果研究组血清sRAGE水平、EOS计数均高于对照组,FEV1低于对照组,差异均有统计学意义(P<0.05)。重度组血清sRAGE水平、EOS计数均高于轻中度组,FEV1低于轻中度组,差异均有统计学意义(P<0.05)。转归不良组血清sRAGE水平、EOS计数均高于转归良好组,FEV1低于转归良好组,差异均有统计学意义(P<0.05)。ROC曲线结果显示,血清sRAGE单独诊断支气管哮喘患者严重程度的曲线下面积(AUC)为0.846,最佳截断值为2.86 ng/mL,EOS计数单独诊断支气管哮喘患者严重程度的AUC为0.874,最佳截断值为0.44×10^(9)/L;FEV1单独诊断支气管哮喘患者严重程度的AUC为0.918,最佳截断值为69.38%,三者联合诊断的AUC(0.965)显著大于三者单独诊断的AUC。血清sRAGE单独诊断支气管哮喘患者结局转归的AUC为0.836,最佳截断值为3.90 ng/mL,EOS计数单独诊断支气管哮喘患者结局转归的AUC为0.809,最佳截断值为0.50×10^(9)/L;FEV1单独诊断支气管哮喘患者结局转归的AUC为0.842,最佳截断值为63.83%;三者联合诊断的AUC(0.941)显著大于三者单独诊断的AUC。结论血清sRAGE水平、EOS计数和FEV1对支气管哮喘病情严重程度及结局转归均具有一定的临床诊断价值,三者联合诊断的价值最高。