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Objective:To investigate the relationship between advanced glycation end products (AGEs) in the lens and type 2 diabetes mellitus.Methods:226 subjects were recruited between August 14 to September 14, 2018 from the Endocrinology Department of Central South University Xiangya Hospital, the Third Hospital of Changsha City, and the Fourth Hospital of Changsha City. The OGTT test, combined with clinical indicators, were used as the gold standard. Subjects were screened for type 2 diabetes using both the lens AGE fluorescence assay and the gold standard. Drawing the receiver operating characteristic (ROC) curve, we calculated the area under the curve (AUC) and its 95% CI and calculated the AGE for the diagnosis of type 2 diabetes. Sensitivity, specificity, Youden index, Kappa value, and its 95% CI, and the optimal cut-off value were determined according to the Youden index. Taking diabetes as the outcome indicator and AGE as the binary indicator, three logistic regression models were constructed. Stratified by age and sub-center, the differences between fasting blood glucose and 2 h postprandial blood glucose were compared between the AGE-negative and AGE-positive groups to determine the relationship between AGE and diabetes. Results:The area under the ROC curve was 0.86(95% CI: 0.81-0.91). According to the Youden index, the optimal cut-off point for AGE was 0.24. At this time, the sensitivity was 82.86(95% CI: 77.81-87.91), the specificity was 77.06(95% CI: 71.43-82.7), the Youden index was 59.92(95% CI: 53.36-66.49), the Kappa value was 79.62(95% CI: 74.22-85.02). Except for the 20-39-year-old group, the fasting blood glucose and 2 h postprandial blood glucose of the AGE-positive group in different age groups, different sub-centers, and the general population were higher than those of the AGE-negative group (all P<0.05). After adjusting for the confounding effects of age, gender, and sub-center (model 3), the relative risk of diabetes in the AGE-positive group was 11.75 times higher than the AGE-negative group (95% CI: 5.61-24.60), all with P<0.001. Conclusion:There was a high correlation between AGE in the lens and the risk of type 2 diabetes. When the cut-off point of AGE is 0.24, it had high sensitivity and specificity and could be used as a practical tool for early screening of type 2 diabetes.
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Objective:To explore the peripheral blood levels changes of soluble glycosylation end product receptor (sRAGE), endotoxin and Toll-like receptor (TLR) in patients with severe acute pancreatitis (SAP) complicated with peritoneal cavity infection, and clarify the relationship between indexes and pathogenetic condition.Methods:The clinical data of 105 patients with SAP in Shanghai Pudong New Area People′s Hospital from January 2017 to December 2020 were retrospectively analyzed. Among them, 28 cases had peritoneal cavity infection (infection group), and 77 cases had peritoneal cavity infection symptoms but undiagnosed (non-infection group). The acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) at admission was recorded. When patients had the suspected symptoms and signs of peritoneal cavity infection, the serum levels of sRAGE, endotoxin, TLR4 and TLR9 were detected by enzyme-linked immunosorbent assay. The correlation between serum levels of sRAGE, endotoxin, TLR-4, TLR-9 and APACHE Ⅱ was analyzed by Pearson analysis; the risk factors of peritoneal cavity infection in SAP patients were analyzed by multivariate Logistic regression; the receiver operating characteristic (ROC) curve was drawn, and the diagnostic efficacy of serum sRAGE, endotoxin, TLR-4 and TLR-9 in peritoneal cavity infection were evaluated in patients with SAP; the Kaplan-Meier survival curve was drawn, and the comparison used log-rank test.Results:The serum sRAGE, endotoxin, TLR-4 and TLR-9 in infection group were significantly higher than those in non-infection group: (822.16 ± 104.51) ng/L vs. (728.09 ± 96.47) ng/L, (62.59 ± 20.11) ng/L vs. (41.62 ± 13.64) ng/L, (45.17 ± 8.54) μg/L vs. (37.34 ± 6.22) μg/L, (26.35 ± 6.73) μg/L vs. (20.02 ± 5.49) μg/L, and there were statistical differences ( P<0.01). Pearson correlation analysis result showed that the serum sRAGE, endotoxin, TLR-4 and TLR-9 were positively correlated with APACHE Ⅱ in patients with SAP ( r = 0.632, 0.556, 0.521 and 0.631; P<0.05). Multivariate Logistic regression analysis result showed that the combined organ function damage, shock, hypoxemia and serum sRAGE, endotoxin, TLR-4 and TLR-9 were independent risk factors of peritoneal cavity infection in patients with SAP ( P<0.05). ROC curve analysis result showed that the area under the curve for the serum sRAGE, endotoxin, TLR-4 and TLR-9 combined diagnosis of peritoneal cavity infection in patients with SAP was the largest, which was 0.910 (95% CI 0.838 to 0.957, P<0.01), with a sensitivity of 82.14% and a specificity of 87.01%. According to the ROC curve cut-off value of serum sRAGE, endotoxin, TLR-4 and TLR-9 (764.58 ng/L, 58.01 ng/L, 40.24 μg/L and 22.61 μg/L), the 28 patients with SAP complicated with peritoneal cavity infection were divided into high levels patients (21, 14, 23 and 22 cases) and low levels patients (7, 14, 5 and 6 cases); Kaplan-Meier survival curve analysis result showed that the 28-day survival rates in patients with high levels of sRAGE, endotoxin, TLR-4 and TLR-9 were significantly lower than those in patients with low levels (61.90% vs. 71.43%, 50.00% vs. 78.57%, 60.87% vs. 80.00% and 59.09% vs. 83.33%), and there were statistical differences ( P<0.05). Conclusions:The serum sRAGE, endotoxin, TLR-4 and TLR-9 have a high combined diagnostic value in SAP complicated with peritoneal cavity infection, and they are all related to the severity of the disease and have a significant impact on survival.
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Objective To observe the protective effect of polypyrimidine bundle-binding proteinrelated splicing factor (PSF) over-expression on RPE cell injury induced by advanced glycation end products (AGEs).Methods The human RPE cells cultured in vitro were divided into three groups:normal control group (N group),blank control group (N + AGEs group),empty vector control group (Vec + AGEs group),and PSF high expression group (PSF + AGEs).group).RPE cells in N group were routinely cultured;RPE cells in N + AGEs group were only transfected but did not introduce any exogenous genes combined with AGEs induction;Vec +AGEs group and PSF + AGEs group were transfected with pcDNA The empty vector or pcDNA-PSF eukaryotic expression plasmid was introduced into RPE cells and induced by AGEs.Except the N group,the other 3 groups of cells were transfected accordingly,and were stimulated with 150 μg/ml AGEs for 72 h after 24 h.HE staining and Hoechst 33258 staining were used to observe the effect of high PSF expression on the morphological changes of RPE cells;ROS level detection was used to analyze the effect of PSF high expression on the ROS expression of RPE cells induced by AGEs;MTT colorimetric method was used to detect the high PSF expression Effects on the viability of RPE cells;Western blot was used to detect the effects of different time and dose of PSF on the expression of heme oxygenase 1 (HO-1).Results HE staining and Hoechst 33258 staining observation showed that the cells in group N were full in shape,the nucleus was round,the cytoplasm was rich,and the staining was uniform;the cells in N + AGEs group and Vec + AGEs group were reduced in size,the eosinophilic staining was enhanced,and the nucleus was densely densely stained.Pyrolysis and even fragmentation;the morphology of cells in the PSF + AGEs group was still full,the cytoplasm staining was more uniform,and the nucleus staining was uniform.The results of MTT colorimetry showed that high expression of PSF can effectively improve the viability of RPE cells,but this effect can be effectively antagonized by ZnPP,and the difference is statistically significant (F=33.26,P＜0.05).DCFH-DA test results showed that compared with the N + AGEs group and Vec + AGEs group,the ROS production in PSF + AGEs group decreased,the difference was statistically significant (F=1 1.94,P＜0.05).Western blot analysis showed that PSF protein upregulated HO-1 expression in a time-and dose-dependent manner.The relative expression level of HO-1 at 24,48,and 72 h after PSF protein was significantly higher than that at 0 h,and the difference was statistically significant (F=164.91,P＜0.05).The relative expression level of HO-1 under the action of 0.1,0.5,1.0,1.5,and 2.0 μg PSF protein was significantly higher than 0.0 μg,and the difference was statistically significant (F=104.82,P＜0.05).Conclusion PSF may inhibit the production of ROS by up-regulating the expression of HO-1,thus protecting the RPE cells induced by AGEs.
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Objective To observe the effect ofpolypyramidine tract binding protein-associated splicing factor (PSF) towards advanced glycation end products (AGEs) induced the apoptosis of Müller cells in vitro.Methods Experimental study.Müller cells were cultured and divided into groups according to the project design,plasmid enhanced green fluorescent protein-PSF were transfected into the cells to achieve the overexpression of PSF Müller cells in vitro,then cells were exposed to AGEs and the Morphological changes were observed by HE staining and Hoechst 33258 staining while the survival rate of cells were detected by MTT assay.The effects of PSF on AGEs-induced Müller apoptosis was measured by Cell Death Detection ELISA kit.Meanwhile,2',7'-diehlorofluorescin diaeetate staining was performed to monitor the protective effects of PSF on AGEs-induced Müller cells ROS.Results The morphology of cells in normal group was full and the cytoplasm staining was uniform.In N+AGEs group and Vec+AGEs group,cell volume decreased,cytoplasm was dense and concentrated,and eosinophilic staining was enhanced.The cell morphology of PSF+AGEs group was still full,with uniform cytoplasm staining and uniform nucleus staining.The viability of N+AGEs group,Vec+AGEs group and PSF+AGEs group were 0.42±0.11,0.35±0.12 and 0.68±0.12.The apoptosis values were 1.08 ± 0.16,0.96± 0.20 and 0.44± 0.08.The intracellular ROS levels were 28 833.67± 3 550.06,28 356.67±4 854.81,186 163.00±382.54.Compared with N+AGEs group and Vec+AGEs group,the cell viability of PSF+AGEs group was significantly improved (F=20.65,P=0.000),ce11 apoptosis value (F=43.43,P=0.000) and intracellular ROS level (F=1 8.86,P=0.000).Conclusion PSF overexpression play a protective role in AGEs-induced apoptosis by inhibiting the production of ROS in Müiller cells.
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Objective To investigate the effects and mechanism of advanced glycosylation end products (AGEs)on endothelial cell senescence and endothelial barrier dysfunction.Methods Human umbilical vein endothelial cells (HUVECs)were isolated and cultured.The cells were randomized into three groups:the control group(normal medium),the bovine serum albumin-treated group(BSA control group)and AEGs group(treated with AEGs-BSA).Senescence of HUVECs were detected by senescence-associated β-galactosidase (SA-beta-Gal)staining.The mRNA and protein expressions of senescence-related genes of p53,p21 and p16 in each group were determined by reverse transcription and real-time PCR(RT-qPCR)and Western blot.Reactive oxygen species(ROS)level was determined by dichlorodihdrofluorescence diacetate (DCFH-DA).The transendothelial electric resistance(TER)were measured by endothelial electric resistance meter.The protein levels of myosin light chain kinase (MLCK),phosphorylated myosin light chain (p-MLC),myosin light chain (MLC)were detected by Western blot.Results Compared with the control group and the BSA control group,the AGEs group showed the significantly increased positive rate of senescence-associated SA-beta-Gal staining (67.30 ± 0.75 ％ vs.7.81 ±0.35 ％ and 7.64 ± 0.91％,respectively,P ＜ 0.01)and the expressions of aging-related genes of p53,p21 and p16 were significantly increased (P ＜ 0.05)There was no significant difference in transendothelial electric resistance(TER)between the control group and theBSAgroup(48.0±6.3 Ω· cm2 vs.42.0±7.8 Ω· cm2,P＞0.05),while TER was lower in the AEGs group than in control group and the BSA group[(27.0±4.2)Ω · cm2 vs.(48.0±6.3)Ω · cm2 and (42.0 ± 7.8) Ω · cm2,P ＜0.01].ROS production had no significant difference between the control group and the BSA group[(38.36 ± 8.55) ％ vs.(41.67 ± 6.93) ％,p ＞ 0.05],while was increased in the AEGs group versus control group and the BSA group[(69.31±8.47)％ vs.(38.36±8.55) ％ and (41.67 ± 6.93) ％,P ＜0.05).The protein expression levels of MLK and p-MLC/MLC were higher in the AGEs group than in the control group and the BSA group(P＜0.05).Conclusions AGEs may lead to endothelial cell senescence and endothelial barrier dysfunction by promoting ROS production and oxidative stress,and by regulating MLCK signaling pathway.
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Objective To investigate the effects of fluoride on protein oxidative damage in rat plasma by measuring oxidative stress levels,advanced glycation end products (AGEs) and advanced oxidation protein products (AOPP).Methods Eighty SPF male 3-week-old Wistar rats weighing (82.34 ± 10.60) g were randomly divided into 4 groups,20 rats in each group.The control group drank distilled water,and the fluoride groups drank distilled water with fluoride concentrations of 25,50 and 100 mg/L,respectively.Rats were allowed to eat and drink freely,and they were sacrificed at 1 month and 3 month,respectively,and samples such as urine,femur and peripheral blood were collected for experiments.Fluoride contents in urine and bone were detected by ion selective electrode method,the superoxide dismutase (SOD) activity was detected by hydroxylamine method,malondialdehyde (MDA) content was detected by thiobarbituric acid (TBA) method,and AGEs and AOPP contents were detected by enzyme linked immunosorbent assay (ELISA).Results For 1 month and 3 months,compared urinary fluoride contents (mg/L:2.088 + 0.638,9.170 ± 2.865,20.094 ± 8.186,54.866 ± 2.866;2.202 ± 1.282,9.112 ± 2.364,21.854 ±8.325,52.513 ± 16.211),and bone fluoride contents (mg/kg:324.985 ± 127.094,846.148 ± 331.861,1 886.601 ±250.140,2 420.971 ± 135.883;417.591 ± 88.324,1 582.243 ± 347.975,2 163.519 ± 614.932,2 755.434 ±265.370) in control group and fluoride concentrations of 25,50 and 100 mg/L groups,the differences were statistically significant (F =88.379,29.225;87.440,33.998,P ＜ 0.05).For 1 month and 3 months,compared SOD activity (U/ml:32.469 ± 5.674,35.931 ± 2.262,36.746 ± 3.994,38.042 ± 4.632;31.027 ± 4.147,30.777 ±4.791,34.148 ± 1.755,36.585 ± 2.860) and AGEs contents (μg/L:26.977 ± 5.285,33.303 ± 6.226,28.021 ±5.946,34.117 ± 6.706;35.681 ± 3.802,33.651 ± 7.214,28.114 ± 4.660,24.330 ± 3.581) in control group and fluoride concentrations of 25,50 and 100 mg/L groups,the differences were statistically significant (F =2.896,5.780;3.565,10.195,P ＜ 0.05).By factorial design anova,there was an interaction between the exposure concentration and exposure time of fluorine and the content of AGEs (F =8.957,P ＜ 0.01).Conclusion Excessive fluoride can affect urinary,bone fluoride contents,SOD activity,AGEs content,suggesting that excessive fluoride may regulate protein expression through direct and indirect oxidative damage pathways,which leading to fluorosis.
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Objective To investigate the regulatory role of cathepsin D (CatD) in the degradation of intracellular advanced glycation end products (AGEs) endocytosed by human dermal fibroblasts (HDFs).Methods Cultured HDFs were treated with 1 μnol/L CA074Me (an inhibitor of CatB and CatL),75 μmol/L pepstatin A (an inhibitor of CatD) and 1 μmol/L MG-132 (an inhibitor of20S proteasome) separately for 4 hours,and then cell counting kit 8 (CCKS) assay and fluorometric assay were performed to determine the cellular viability and protease activity,respectively.The cells in the CA074Me group,pepstatin A group and MG-132 group were additionally treated with AGE-bovine serum albumin (BSA) for 8 hours,and the cells in the blank control group were treated with phosphate-buffered saline (PBS) alone.After 8-hour cultivation,the cells in the above groups were subsequently reincubated with fresh culture medium containing the corresponding inhibitors for 24 hours.Then,flow cytometry was performed to assess the mean fluorescence intensity of intracellular AGE-BSA at different time points.Some other HDFs were treated with 37.5,75 and 150 μmol/L pepstatin A and PBS separately for 4 hours,and then the cells in the 4 groups were treated with 200 mg/L AGE-BSA for 8 hours,followed by the removal of AGE-BSA from the medium and the treatment with 37.5,75 and 150 μmol/L pepstatin A and PBS respectively.Enzyme-linked immunosorbent assay (ELISA) was conducted to measure the mean concentration of intracellular AGE-BSA at different time points,and the degradation rate of AGE was calculated.Some HDFs were divided into 3 groups:blank control group receiving no treatment,NC group transfected with an empty vector,and CatD group transfected with a CatD-overexpressing lentiviral vector.Fluorescence microscopy was conducted to estimate the transfection efficiency.Reverse transcription-PCR,Western blot analysis and fluorometric assay were performed to determine the mRNA and protein expression,and activity of CatD respectively.Then,the cells in the above 3 groups were incubated with AGE-BSA for 8 hours,followed by the removal of AGE-BSA from the medium and the treatment with fresh culture medium.The detection methods were same as the above experiment,and the degradation rate was calculated.Results The cellular proliferative activity in the 1-μmol/L CA074Me group,75-μmnol/L pepstatin A group and 1-μ mol/L MG-132 group was more than 90％,and there was no significant difference between the 3 groups and the control group (100％,F =1.525,P ＞ 0.05).Twenty-four hours after the removal of AGE-BSA from the medium,the fluorescence intensities of intracellular AGE-BSA in the CA074Me + AGE-BSA group (275.00 ± 10.15) and MG-132 + AGE-BSA group (259.00 ± 11.14) significantly decreased compared with those at the 8-hour time point (295.00 ± 6.56 and 285.67±8.74 respectively;paired t test,t =4.778,6.154 respectively,both P ＜ 0.05),while no significant difference was observed in the fluorescence intensities of intracellular AGE-BSA in the pepstatin A + AGE-BSA group between the 8-hour time point and 32-hour time point (P ＞ 0.05).The degradation rates of intracellular AGE-BSA within 24 hours in the 37.5,75 and 150 μmol/L pepstatin A groups were 9.64％ ± 1.27％,5.62％ ± 0.47％ and 3.21％ ± 0.73％ respectively;there were significant differences among the 3 groups (F =45.876,P ＜ 0.05),and the degradation rate significantly decreased along with the increase of pepstatin A concentration (P ＜ 0.05).Fluorescence microscopy showed no fluorescent cells in the blank control group,while the NC group and CatD group both showed a high proportion (＞ 80％) of fluorescent cells.The mRNA and protein expression as well as the activity of CatD were significantly higher in the CatD group than in the blank control group and NC group (all P ＜ 0.05).The CatD + AGE-BSA group showed a significantly higher degradation rate of intracellular AGE-BSA within 24 hours compared with the AGE-BSA group and NC + AGE-BSA group (both P ＜ 0.05).Conclusion CatD can promote the degradation of intracellular AGE-BSA endocytosed by HDFs.
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Objective To determine the expression of cathepsin D and advanced glycation end products (AGEs)in skin tissues from patients of different ages or skin tissues with different degrees of sun exposure,to evaluate their correlation,and to preliminarily investigate the role of cathepsin D in the degradation and accumulation of AGEs in photoaged skin.Methods Skin tissues were collected from sunexposed and sun-protected body sites in patients aged 15-20 years,35-40 years,55-60 years or 75-80 years.These skin tissues were divided into 8 groups according to age of patients and degrees of sun exposure,and there were 6 specimens in each group.Immunohistochemical and immunofluorescent methods were used to measure the expression of cathepsin D and AGEs in the skin tissues.Statistical analysis was carried out by factorial design analysis of variance,Wilcoxon rank sum test and Kruskal-Wallis rank sum test for analyzing associations of the expression of cathepsin D and AGEs with age and sun exposure,as well as by Pearson correlation analysis for assessing the correlation between cathepsin D expression and AGEs expression.Results Immunohistochemical study showed that the expression of cathepsin D markedly decreased along with the increase of age,but the accumulation of AGEs gradually increased along with the increase of age.In the same age group,the cathepsin D expression was lower in the sun-exposed skin tissues than in the sun-protected skin tissues,while the accumulation of AGEs was more in the sun-exposed skin tissues than in the sun-protected skin tissues.Factorial design analysis of variance showed that sun exposure could decrease the expression of cathepsin D (F =58.70,P ＜ 0.001),but increase the accumulation of AGEs (F =158.18,P ＜ 0.001).Moreover,the increase of age could lead to decreased expression of cathepsin D (F =79.49,P ＜ 0.001),and increased expression of AGEs (F =106.06,P ＜0.001).Compared with the sun-protected skin tissues,the sun-exposed skin tissues in all the age groups showed significantly lower absorbance value of cathepsin D (35-40 years:0.020 ± 0.005 vs.0.032 ± 0.005;55-60 years:0.012 ± 0.004 vs.0.026 ± 0.002;75-80 years:0.002 ± 0.001 vs.0.013 ± 0.004;all P ＜0.001),but higher absorbance value of AGEs (35-40 years:0.030 ± 0.008 vs.0.010 ± 0.003;55-60years:0.066 ± 0.010 vs.0.021 ± 0.004;75-80 years:0.085 ± 0.015 vs.0.035 ± 0.009;all P ＜ 0.001)except the age group of 15-20 years.No matter whether the skin tissues were sun-exposed or sunprotected,there were significant differences in the expression of cathepsin D and AGEs among different age groups (all P ＜ 0.001).The results of double immunofluorescence staining were similar to those of immunohistochemical study.Pearson correlation analysis showed that the expression of cathepsin D in the sun-exposed skin tissues was highly negatively correlated with the accumulation of AGEs (r =-0.915,P ＜0.05),while they were moderately negatively correlated in the sun-protected skin tissues (r =-0.730,P ＜0.05).Conclusions Along with the increase of age,the expression of cathepsin D in skin tissues decreased,but the expression of AGEs increased.In the sun-protected skin tissues,the expression of cathepsin D was moderately negatively correlated with the expression of AGEs,while they were highly negatively correlated in the sun-exposed skin tissues,suggesting that cathepsin D may play an important role in the degradation and accumulation of AGEs in photoaged skin.
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Objective To investigate the effect and its mechanism of D-pinitol on advanced glycation end products(AGEs)-induced proliferation and migration in mouse aortic vascular smooth muscle cells (VSMC).Methods VSMCs were isolated from mouse aorta and cultured in vitro.Effects of different concentrations of D-pinitol on proliferation and migration of VSMCs were observed by using the AGEs-induced glycosylation injury model of VSMCs.Cell proliferation and migration were detected by CCK-8 assay and cell scratch,respectively.The protein expression levels of transforming growth factor-β1(TGF-β1),p-smad2,p-smad3 and asporin were determined by Western blot.Results Compared with control group,AGEs group showed the increased protein expression levels of asporin,TGF-β1,p-smad2 and p-smad3 (40.06 ± 4.50 vs.17.47 ± 0.57),(55.25 ± 2.07 vs.14.42± 2.07),(0.97 ± 0.02 vs.0.47 ± 0.02),(0.45±0.01 vs.0.26 ± 0.02),all P＜ 0.01.Compared to AGEs group,D-pinitol group could inhibit the cell proliferation and migration and cause dose-dependent decreases of protein expressions of TGF-β1,p-smad2,p-smad3 and asporin(P ＜ 0.05 or P＜0.01).Conclusions D-pinitol can inhibit AGEs-induced cell proliferation and migration in mouse aortic VSMCs.Asporin may participate in the VSMCs extracellular matrix remodeling via TGF-β/smad pathway.
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Abstract Introduction: Chronic kidney disease (CKD) is associated with high morbidity and mortality rates, main causes related with cardiovascular disease (CVD) and bone mineral disorder (CKD-BMD). Uremic toxins, as advanced glycation end products (AGEs), are non-traditional cardiovascular risk factor and play a role on development of CKD-BMD in CKD. The measurement of skin autofluorescence (sAF) is a noninvasive method to assess the level of AGEs in tissue, validated in CKD patients. Objective: The aim of this study is analyze AGEs measured by sAF levels (AGEs-sAF) and its relations with CVD and BMD parameters in HD patients. Methods: Twenty prevalent HD patients (HD group) and healthy subjects (Control group, n = 24), performed biochemical tests and measurements of anthropometric parameters and AGEs-sAF. In addition, HD group performed measurement of intact parathormone (iPTH), transthoracic echocardiogram and radiographies of pelvis and hands for vascular calcification score. Results: AGEs-sAF levels are elevated both in HD and control subjects ranged according to the age, although higher at HD than control group. Single high-flux HD session does not affect AGEs-sAF levels. AGEs-sAF levels were not related to ventricular mass, interventricular septum or vascular calcification in HD group. AGEs-sAF levels were negatively associated with serum iPTH levels. Conclusion: Our study detected a negative correlation of AGEs-sAF with serum iPTH, suggesting a role of AGEs on the pathophysiology of bone disease in HD prevalent patients. The nature of this relation and the clinical application of this non-invasive methodology for evaluation AGEs deposition must be confirmed and clarified in future studies.
Resumo Introdução: A doença renal crônica (DRC) apresenta elevadas taxas de morbidade e mortalidade, sendo a doença cardiovascular (DCV) e o distúrbio mineral e ósseo da DRC (DMO-DRC) complicações frequentes. As toxinas urêmicas, dentre elas os produtos finais da glicação avançada (AGEs), são fatores de risco cardiovascular não tradicionais e se encontram envolvidas no desenvolvimento do DMO-DRC na DRC. A medida da autofluorescência da pele (sAF) é método não invasivo para quantificação do acúmulo tecidual de AGEs validado em pacientes portadores de DRC. Objetivos: O objetivo deste estudo é avaliar as relações entre os AGEs medidos por sAF (AGEs-AF) e parâmetros de DCV e DMO-DRC em pacientes em hemodiálise (HD). Métodos: 20 pacientes em HD (grupo HD) e 24 indivíduos hígidos (grupo controle) foram submetidos à análise bioquímica sérica, medidas antropométricas e de sAF. O grupo HD realizou medida de hormônio intacto da paratireoide (PTHi), ecocardiograma transtorácico e radiografias de pelve e mãos para pesquisa de calcificação vascular. Resultados: Os níveis de AGEs-sAF foram elevados para a idade nos grupos HD e controle, porém mais elevados no grupo HD. Sessão única de HD de alto-fluxo não afetou os níveis de AGEs-sAF. Os níveis teciduais de AGEs não se correlacionaram com massa ventricular, espessura de septo interventricular ou calcificação vascular no grupo HD. Os níveis de AGEs-sAF se correlacionaram negativamente com os níveis séricos de PTHi. Conclusão: Nosso estudo detectou correlação negativa entre os níveis de AGEs-sAF e os níveis séricos de PTHi, sugerindo que os AGEs estejam envolvidos na fiosiopatologia da doença óssea em pacientes em HD. A natureza desta relação e a aplicação clínica deste método não invasivo de avaliação do acúmulo tecidual de AGEs deve ser confirmada e elucidada por estudos futuros.
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Humans , Male , Female , Adult , Chronic Kidney Disease-Mineral and Bone Disorder/metabolism , Skin/metabolism , Glycation End Products, Advanced/metabolism , Chronic Kidney Disease-Mineral and Bone Disorder/diagnostic imaging , Skin/diagnostic imaging , Pilot Projects , Cross-Sectional Studies , Glycation End Products, Advanced/analysis , Optical ImagingABSTRACT
Objective@#To observe the correlation between Nε-carboxymethyl-Lysine (CML), the main component of advanced glycation end products and the calcification of the anterior tibial artery plaque in patients with diabetic foot post foot amputation.@*Methods@#Sixty patients hospitalized for foot amputation operation due to diabetic foot from June 2012 to June 2016 in the Department of Orthopedics, Affiliated Hospital of Jiangsu University were prospectively recruited.The patients were categorized into mild stenosis (0<stenosis<50%, n=20), moderate stenosis (50%≤stenosis<70%, n=20) and severe stenosis (70%≤stenosis≤100%, n=20) based on the color Doppler ultrasound assessed severity of anterior tibial artery stenosis.The baseline clinical data of patients were collected and anterior tibial artery was isolated.Then, HE staining, O-Cresolphthalein Complexone method, enzymic method and ELASA analysis were then performed to detect the evolution of calcification, arterial calcium content, alkaline phosphatase activity and serum CML concentration, respectively.@*Results@#The results from both color Doppler ultrasound scan before amputation and HE staining after amputation showed that echo intensity as well as spotty blue calcium particles of anterior tibial artery plaque increased significantly in proportion to degree of stenosis and destructed elastic plate of the arterial wall was evidenced in patients with severest stenosis.The content of calcium ((2.3±0.9), (3.9±1.3), (6.6±1.7) μmol/mg, respectively, P<0.001), ALP activity ((102.4±39.4), (202.3±73.4), (483.7±117.9) U/mg, respectively, P<0.001) and serum CML level ((28.9±4.4), (37.9±5.3), (57.3±7.1)μg/L, respectively, P<0.001) increased significantly in proportion to stenosis severity.Pearson correlation analysis showed that serum CML level was positively correlated with the content of calcium (r=0.749, P<0.001) and ALP activity (r=0.923, P<0.001), respectively.@*Conclusions@#Serum CML level is positively correlated with the calcification of anterior tibial arterial plaque in patients with diabetic foot and could be used to evaluate the calcification of anterior tibial arterial plaque and stenosis degree of anterior tibial arterial in these patients.
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Background The deposition of advanced glycation end products (AGEs) in lens is the risk factor of diabetic complications.Researches revealed that AGEs has autofluorescence.Crystallin is a longevity protein.AGEs accumulation is probably associated with diabetic retinopathy (DR).Objective This study was to evaluate the association of AGEs autofluorescence intensity with diabetes and with DR.Methods A cross-sectional study was carried out under the approval of Ethic Committee of Shenyang He Eye Hospital and informed consent of each patient.One hundred eyes of 100 patients with age-related cataract aged 50-70 years were included in He Eye Hospital from September to December 2015.The patients were divided into non-diabetes group (40 patients) and diabetes group (60 patients),and then the patients in diabetes group were subdivided into non-DR (NDR) group,non-proliferating DR (NPDR) group and proliferating DR (PDR) group according to the DR grading criteria,20 patients for each.Glycosylated henoglobin A1c (HbA1c) and fasting plasma glucose (FPG) were detected for each subject,and the lens autofluorescence was assayed with lens fluorescence biomicroscope (Clearpath DS120).The association of lens autofluorescence intensity with serum HbA1c level or DR severity was analyzed.Results The age and diabetes course were matched among the non-diabetes group,NDR group,NPDR group and PDR group (F=2.587,2.899,both at P＞0.05),and the FBS and HbA1c level were evidently higher in the NDR group,NPDR group and PDR group than those in the non-diabetes group (all at P＜0.01).The autofluorescence intensity of lens was (0.159±0.032),(0.256±0.024),(0.319 ±0.013) and (0.394±0.035) cd in the non-diabetes group,NDR group,NPDR group and PDR group,respectively,showing a significant difference among the groups (F =90.265,P =0.000).The autofluorescence intensity of lens in the NDR group,NPDR group and PDR group was significantly increased in comparison with the non-diabetes group and the autofluorescence intensity of lens was gradually increased with the severity of DR (all at P＜0.01).A positive linear correlation was found between autofluorescence intensity of lens and serum HbA1 c level in diabetes patients (r =0.654,P ＜ 0.05).Conclusions The autofluorescence intensity of AGEs in lens appears to be associated with the severity of DR and HbA1 c.The autofluorescence intensity of AGEs in the lens of diabetes patient is probably one of the evaluation indexes of early stage of DR.
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Objective:To explore the expression and clinical meaning of plasma soluble receptor of advanced glycation end product (sRAGE) in patients with gestational hypertension heart disease.Methods:Our research included in 2 groups:Observation group,n=57 patients with gestational hypertension heart disease who were diagnosed and treated in our hospital from 2013-06 to 2016-07;Control group,n=57 healthy pregnant women at the same period of time.Based on cardiac function,the patients in Observation group was divided into 4 subgroups as NYHA Ⅰ,Ⅱ,Ⅲ and ⅣV;according to 28-day surviving,the patients were divided into another set of 2 subgroups as Survival subgroup,n=49 and Death subgroup,n=8.Plasma levels of sRAGE and BNP were examined within 24h of admission.The changes of sRAGE were compared among different groups and the impact of sRAGE on gestational hypertension heart disease prognosis was analyzed.Results:Compared with Control group,Observation group had increased plasma sRAGE (939.2±184.3) pg/mL vs (467.3±116.2) pg/mL,P＜0.05;sRAGE level was positively related to NYHA grading (r=0.76,P＜0.001),sRAGE had an elevating trend upon NYHA grade increasing accordingly,P＜0.05.Compared with Survival subgroup,Death subgroup had increased plasma sRAGE (1647.6±249.7) pg/mL vs (776.9±146.2) pg/mL,P＜0.05.The areas of sRAGE and BNP under ROC curve were 0.91 and 0.88 respectively,P＜0.05.Conclusion:Plasma sRAGE was increased in patients with gestational hypertension heart disease;it was related to severity of heart failure (HF) and could be used for predicting the early prognosis in HF patients.
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Objective:To explore the expression and clinical meaning of plasma soluble receptor of advanced glycation end product (sRAGE) in patients with gestational hypertension heart disease.Methods:Our research included in 2 groups:Observation group,n=57 patients with gestational hypertension heart disease who were diagnosed and treated in our hospital from 2013-06 to 2016-07;Control group,n=57 healthy pregnant women at the same period of time.Based on cardiac function,the patients in Observation group was divided into 4 subgroups as NYHA Ⅰ,Ⅱ,Ⅲ and ⅣV;according to 28-day surviving,the patients were divided into another set of 2 subgroups as Survival subgroup,n=49 and Death subgroup,n=8.Plasma levels of sRAGE and BNP were examined within 24h of admission.The changes of sRAGE were compared among different groups and the impact of sRAGE on gestational hypertension heart disease prognosis was analyzed.Results:Compared with Control group,Observation group had increased plasma sRAGE (939.2±184.3) pg/mL vs (467.3±116.2) pg/mL,P＜0.05;sRAGE level was positively related to NYHA grading (r=0.76,P＜0.001),sRAGE had an elevating trend upon NYHA grade increasing accordingly,P＜0.05.Compared with Survival subgroup,Death subgroup had increased plasma sRAGE (1647.6±249.7) pg/mL vs (776.9±146.2) pg/mL,P＜0.05.The areas of sRAGE and BNP under ROC curve were 0.91 and 0.88 respectively,P＜0.05.Conclusion:Plasma sRAGE was increased in patients with gestational hypertension heart disease;it was related to severity of heart failure (HF) and could be used for predicting the early prognosis in HF patients.
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OBJECTIVE To analyze the effect of advanced glycation end products(AGEs) on apoptosis of cultured mouse spiral ganglion cells(SGCs) and expression of receptor of AGEs(RAGE). To explore the pathway of AGEs in promoting apoptosis of SGCs. And to explore the possible mechanism of neural presbycusis. METHODS The effect of AGEs on apoptosis of SGCs was studied by Tunel technique and fluorescence microscope. The expression of RAGE mRNA was assayed by Real time RT-PCR. RESULTS AGEs induced apoptosis of cultured SGCs. The effects were dose-dependent and time-dependent. Meanwhile RAGE mRNA expression was enhanced in apoptosis cells. CONCLUSION AGEs induced apoptosis in SGCs,which may be mediated by RAGE. And this may be one of the mechanisms of neural presbycusis.
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Objective To investigate the effects of advanced glycation end products(AGE)on the expressions and activity of cathepsin D(CatD)in ultraviolet A(UVA)?irradiated human dermal fibroblasts. Methods Human dermal fibroblasts were isolated and harvested from the circumcised foreskin of children, and subjected to a primary culture. CCK?8 assay was performed to screen non?cytotoxic concentrations of AGE?bovine serum albumin (BSA). Some fibroblasts were incubated with 50, 100 and 300 mg/L AGE?BSA separately for 24 hours, with untreated cells as the control group. Then, reverse transcription(RT)?PCR, Western?blot analysis and a fluorimetric assay were performed to measure the mRNA and protein expressions as well as activity of CatD, respectively. Some fibroblasts were classified into six groups: control group receiving no treatment, AGE?BSA group and BSA group treated with the highest non?cytotoxic concentration of AGE?BSA and the same concentration of BSA respectively for 24 hours, UVA group irradiated by 10 J/cm2 UVA, UVA?AGE?BSA group and UVA?BSA group treated with AGE?BSA and BSA at the above non?cytotoxic concentration respectively for 24 hours both before and after UVA radiation at 10 J/cm2. After the treatments, RT?PCR, Western?blot analysis and a fluorimetric assay were conducted to detect mRNA and protein expressions and activity of CatD respectively. Results AGE?BSA of 50- 200 mg/L exhibited no obvious influence on cellular proliferation of fibroblasts. The fibroblasts incubated with AGE?BSA of 50, 100 and 200 mg/L showed a significant increase in the mRNA expression(0.267 ± 0.007, 0.348 ± 0.007, and 0.418 ± 0.006 respectively), protein expression (1.403 ± 0.181, 2.233 ± 0.090 and 2.477 ± 0.111 respectively), and activity(1.760 ± 0.080, 2.330 ± 0.060 and 2.890 ± 0.080 respectively)of CatD compared with the control group(mRNA:0.161 ± 0.006;protein:0.903 ± 0.200;activity:1.100 ± 0.090, all P < 0.05). AGE?BSA increased CatD expressions and activity in a dose?dependent manner. The mRNA and protein expressions as well as activity of CatD were significantly higher in the UVA group than in the control group (mRNA expression: 0.480 ± 0.005 vs. 0.155 ± 0.005; protein expression: 2.583 ± 0.199 vs. 0.920 ± 0.235;activity:2.970 ± 0.110 vs. 1.110 ± 0.040, all P<0.05), but significantly lower in the UVA?AGE?BSA group than in the UVA group(mRNA expression:0.394 ± 0.008 vs. 0.480 ± 0.005;protein expression:2.070 ± 0.125 vs. 2.583 ± 0.199;activity: 2.560 ± 0.060 vs. 2.970 ± 0.110, all P < 0.05). Conclusion AGEs could increase CatD expressions and activity in human dermal fibroblasts not receiving UVA irradiation, but inhibit their increase in UVA?induced human dermal fibroblasts.
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Objective To investigate the association of genetic polymorphism in the advanced glycation endproduct (RAGE) gene with genetic susceptibility to ischemic stroke.Methods A case-control study was conducted with 124 ischemic stroke patients serving as the stroke group and 125 healthy volunteers serving as the control group.Three common polymorphisms in the RAGE gene,82G/S (rs2070600),-429T/C (rs1800625) and-374T/A (rs1800624),were genotyped by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RELP) method.The association between the three polymorphisms and genetic susceptibility to ischemic stroke was further analyzed by logistic regression analysis.Results The body mass index (BMI),diastolic blood pressure,blood lipids levels,and the percentage of smokers were significantly higher in the stroke group than in the control group (each P＜0.05).At the 82 position,the frequencies of the genotypes GG,GS and SS were 10％,48％ and 42％,respectively.At the 429 position,the frequencies of the genotypes TT,TC and CC were 15％,44％ and 41％,respectively.At the 374 position,the frequencies of the genotypes TT,TA and AA were 8％,41％ and 50％,respectively.After adjusting for confounding factors including age,gender,smoking and blood pressure,the genotype frequencies of SS and S alleles at 82G/S of the RAGE gene were higher in the stroke group than in control group (SS:OR=2.14,95％ CI:1.37-3.51;S:OR=1.96,95％ CI:1.24-3.34),while no significant differences of genotype frequencies of the 429T/C and-374T/T polymorphisms were observed between the stroke group and the control group (each P＞0.05).Conclusions The 82G/S gene polymorphism of the RAGE gene has correlations with ischemic stroke in the Henan Han population,with the S allele as the risk factor for ischemic stroke.However,no clear association between the 429T/C or-374T/A polymorphisms of the RAGE gene and ischemic stroke is identified in the Han population.
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Las evidencias que apoyan la hiperglucemia crónica como causante de una serie de complicaciones macro y mi-crovasculares son abrumadoras. Las alteraciones fisiopatológicas que se derivan de esta patología van más allá del significado de niveles elevados de glucosa, como consecuencia de una secreción adecuada de insulina o una resistencia de los tejidos al ingreso de glucosa a las células. Las consecuencias de estos niveles elevados de glucosa por tiempo prolongado, en última instancia conducen a la glicación de las proteínas, cuyas consecuencias es un funcionamiento deficiente, además de la formación de productos finales de glicación avanzada. La evaluación de la hemoglobina glicosilada, o la albumina glicada son indicadoras del tiempo que llevan las proteínas expuestas a altas concentraciones de glucosa o al estado glicémico del paciente, pero también intervienen en complicaciones a largo plazo como la nefropatía diabética. La consecuencia de estas proteínas glicadas y la formación de produc-tos avanzados de glicación es el mal funcionamiento de órganos vitales, envejecimiento y desarrollo de enferme-dades degenerativas como el Alzheimer.
The evidence supporting chronic hyperglycemia as the cause of a series of macro and microvascularcomplications are devastating. Pathophysiologic changes that result from this condition go beyond the meaning of high levels of glucose, as a result of inadequate insulin secretion or tissue resistance to the entry of glucose into cells. The consequences of these high glucose levels for prolonged periods, ultimately lead to the glycation of proteins, the consequences of a malfunction, in addition to the formation of advanced glycation end products. The evaluation of glycated hemoglobin, glycated albumin or time are indicative proteins bearing exposed to high concentrations of glucose or glycemic condition of the patient, but also involved in long-term complications as diabetic nephrop-athy. The consequence of these glycated proteins and the formation of advanced glycation end products are the malfunction of vital organs aging and development of degenerative diseases like Alzheimer's
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Humans , Pathology , Proteins , Diabetes MellitusABSTRACT
Background Receptor for advanced glycation end products (RAGE) play an important role in the process of type 2 diabetes and its microvascular complications.RAGE gene Gly82Ser exists polymorphism,but the correlation of gene polymorphism and diabetic retinopathy (DR) needs further research.Objective This study was to investigate the association of Gly82Ser polymorphism of RAGE gene with DR in Han people of Wuxi area with type 2 diabetic mellitus.Methods One hundred and eighty-five patiens with type 2 diabetes were included in Wuxi district from March 2013 to February 2014.The patients were divided into non DR (NDR) group (93 cases) and DR group (92 cases) according to the DR International Clinical Classification Criteria in 2002,and 120 healthy subjects were included at the same time as the control.All of the subjects received eye examinations,body mass index (BMI) and blood pressure measurement as well as laboratory tests,including blood biochemical indexes,blood lipids and fasting blood glucose levels.The peripheral blood of 3 ml was collected from each subject,and the genotype and allelic frequencies were assayed by PCR-direct sequencing.This study protocol was approved by Ethic Committee of Nanjing Medical University.Written informed consent was obtained from each subject prior to any medical examination.Results The course was significantly longer in the DR group than that in the NDR group (t =2.25,P =0.01).There were two alleles of G and A in the RAGE gene Gly82Ser locus in all the subjects and the distribution of single nucleotide polymorphism was in accordance with Hardy-Weinberg equilibrium (DR group:x2 =0.51,P =0.48;NDR group:x2 =1.38,P =0.24;healthy control group:x2 =0.20,P =0.24).The AA genotype frequency of the subjects in DR group,NDR group and healthy control group were respectively 6.5％,3.2％ and 2.5％,and AA genotype frequency in DR group was higher than that of the NDR group and healthy control group,showing significant intergroup differences (x2 =5.146,P =0.023;x2 =5.039,P =0.037).The distribution of A allele frequency in the DR group was significantly higher than that of NDR group and healthy control group (x2=5.494,P=0.019;x2 =5.235,P =0.023),and the frequencies of G allele and GG genotype in the DR group were lower than those of the NDR and the healthy control group (GG:x2 =4.260,P =0.039;x2 =4.794,P =0.027;G:x2 =5.309,P =0.021;x2 =5.476,P=0.032).No significant differences were seen in the frequencies of genotype and allele of subjects between the NDR group and the healthy control group (AA:x2 =5.346,P=0.127;GG:x2 =6.981,P=0.137;A:x2 =5.618,P =0.082;G:x2 =4.860,P =0.088).Conclusions The Gly82Ser polymorphism of RAGE gene is associated with the pathogenesis of DR in Han population with type 2 diabetes and A allele may be a risk factor of DR.
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Objective To determine the association of-429T/C and G1704T polymorphisms in the receptor for advanced glycation end products gene with proliferative diabetic retinopathy (PDR).Methods Case-control study.From the Beijing Desheng Diabetic Eye Study cohort of 1467 patients with type 2 diabetes mellitus (T2DM),a total of 97 patients with PDR and 105 diabetic patients without retinopathy (DWR,duration of diabetes 15 years) were included for this study.Questionnaires were collected and general ophthalmologic examinations were performed.Biochemical analysis was conducted.DNA was extracted from peripheral venous blood.The-429T/C and G1704T single nucleotide polymorphisms were detected by the means of PCR-restrication fragment length polymorphisms.Results The frequency distribution of-429T/C in DWR group was 81.0％ in TT,16.1％ in TC,2.9％ in CC.The frequency distribution of-429T/C in PDR group was 77.3％ in TT,20.6％ in TC,2.1％ in CC.There was no significant statistical difference between the two groups (x2 =0.40,P＞0.05).Frequency of the-429T/C minor allele C in the DWR and PDR group were 11.0％ and 12.4％,respectively,with no significant statistical difference between the two groups (x2 =0.20,P＞0.05).The frequency distribution of G1704T in DWR group was 66.7％ in GG,29.5％ in GT,3.8％ in TT.The frequency distribution of G1704T in PDR group was 78.4％ in GG,21.6％ in GT.There was no significant statistical difference between the two groups (x2 =3.44,P＞0.05).Frequency of the G1704T minor allele T in the DWR and PDR group were 18.6 ％ and 10.8 ％,respectively,in which significant difference was found within the two groups (x2 =4.79,OR=1.88,95％CI:1.06-3.33,P＜0.05).Conclusions G1704T polymorphism is associated with PDR presence and 1704G allele may increase the risk of PDR.