Objective To study the expression of receptor of advanced glycation end products (RAGE) in autogenous vein graft of streptozotocin induced diabetic rats and the inhibitory effects of aminoguanidine on intimal hyperplasia. Methods Sixty male Sprague-Dawley rats were randomly divided into three groups: aminoguanidine group, distilled water group and control group. Autogenous vein graft models were established in all groups. Streptozotocin was injected into abdominal cavity to induce diabetes in both aminoguanidine group and distilled water group, and they were intragastric administrated with aminoguanidine or distilled water, respectively before and after transplantation. Specimens were collected from autogenous vein graft 7 days and 14 days after surgery to undergo histological examination. At the same time, the level of serum advanced glycation end products (AGE) was tested. Western blotting and immunohistochemistry were used to detect the protein expression of RAGE and NF-κB p65. RAGE and NF-κB p65 mRNA were measured by reverse transcription-PCR. Results The mRNA and protein expressions of RAGE, NF-κB p65, the level of serum AGE and the intimal thickness of vein graft in distilled water group increased in comparison with those in control group 7 days and 14 days after surgery （P＜0.05). The level of serum AGE, mRNA and protein expressions of NF-κB p65 and the intimal thickness of vein graft in aminoguanidine group were lower than those in distilled water group （P＜0.05), and showed no significant difference compared with control group （P＞0.05). Conclusion The over-expression of RAGE in vein graft activats NF-κB in streptozotocin-induced diabetic rat, which has a close relation with intimal hyperplasia. Aminoguanidine can block the binding of AGE and RAGE by inhibiting the production of AGE, which will prevent intimal hyperplasia of vein graft.
ObjectiveTo observe the protective effect of polypyrimidine bundle-binding protein-related splicing factor (PSF) over-expression on RPE cell injury induced by advanced glycation end products (AGEs).MethodsThe 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).ResultsHE 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=11.94, P＜0.05). Western blot analysis showed that PSF protein up-regulated 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).ConclusionPSF may inhibit the production of ROS by up-regulating the expression of HO-1, thus protecting the RPE cells induced by AGEs.
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),atotal 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 (χ2=0.40, P > 0.05). Frequency of the -429T/C minor alleleCin the DWR and PDR group were 11.0% and 12.4%, respectively, with no significant statistical difference between the two groups (χ2=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 (χ2=3.44, P > 0.05). Frequency of the G1704T minor alleleTin the DWR and PDR group were 18.6% and 10.8%, respectively, in which significant difference was found within the two groups (χ2=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.
Objective To observe the value of serum soluble receptor of advanced glycation endproducts (sRAGE) combined with lung function and high resolution lung CT (HRCT) in predicting the risk of chronic obstructive pulmonary disease (COPD) developing non-small cell lung cancer (NSCLC). Methods From January 2019 to June 2021, 140 patients with COPD combined with NSCLC, 137 patients with COPD, and 133 patients with NSCLC were enrolled in the study from the People's Hospital of Ningxia Hui Autonomous Region. General data, clinical symptoms, pulmonary function indexes and HRCT emphysema indexes (EI) were collected. Serum sRAGE levels of these patients were measured by enzyme linked immunosorbent assay. Clinical characteristics of patients with COPD complicated with NSCLC were analyzed. Serum sRAGE, lung function and lung HRCT were combined to evaluate the correlation between the degree of emphysema and the occurrence of NSCLC in COPD, and receiver operator characteristic (ROC) curve analysis was performed for diagnostic efficiency. Results Compared with NSCLC group, COPD combined with NSCLC group had higher proportion of male patients, higher proportion of elderly patients, higher smoking index, and higher proportion of squamous cell carcinoma (P<0.05). FEV1 and FEV1%pred in COPD combined with NSCLC group were significantly lower than those in COPD group and NSCLC group. The Goddard score and EI values of emphysema were significantly increased (P<0.05). Serum sRAGE was significantly lower than that of COPD group and NSCLC group (P<0.05). Serum sRAGE level was positively correlated with FEV1%pred (r=0.366, P<0.001) and FEV1/FVC (r=0.419, P<0.001), and negatively correlated with Goddard score (r=–0.710, P=0.001) and EI value (r=–0.515, P<0.001). Binary multi-factor logistic regression analysis showed that age, smoking index, EI, Goddard score, RV/TLC were positively correlated with the risk of COPD developing NSCLC, while FEV1%pred, FVC, FEV1/FVC and serum sRAGE were negatively correlated with the risk of COPD developing NSCLC. ROC curve results showed that the area under the curve (AUC) of single diagnosis of sRAGE was 0.990, and the optimal cut-off value of 391.98 pg/mL with sensitivity of 93.3% and specificity of 89.7%. The AUC of sRAGE combined with age, smoking index, EI, Goddard score, FEV1%pred, FVC, FEV1/FVC, RV/TLC was 1.000 with sensitivity of 96.7%, specificity of 96.6%, and Yoden index of 0.933. Conclusion The combination of serum sRAGE, lung function and HRCT emphysema score can improve prediction of NSCLC occurrence in COPD.
Objective To investigate the effects of advanced glycation endproducts (AGEs) on proliferation of pericytes of bovine retinal capillary vessels and expression of transforming growth factor beta;(TGF-beta;). Methods The proliferation of pericytes detected by methyl thiazolyl tetrazolium (MTT) colorimetric assay, cellular cycle of pericytes was analyzed by flow cytometry was used to analyze cell, and TGF-beta; protein expression of pericytes was observed by immunofluorescent staining. Results AGEs inhibited the proliferation of pericytes of bovine retinal capillary vessels, stopped the cellular cycle of pericytes in synthesis phase (S phase), increased the number of apoptotic cells obviously (Plt;0.01), and promoted the expression of TGF-beta; proteinof perycytes. Conclusions AGEs may promote the apoptosis of pericytes by inhibiting the proliferation of pericytes to lead the decrease of pericytes number, and may accelerate diabetic retinopathy by promoting the expression of TGF-beta; protein of pericytes. (Chin J Ocul Fundus Dis, 2006, 22: 20-23)
【摘要】 目的 观察晚期糖基化终产物（advanced glycosylation end prodrcts，AGE）对人结肠癌细胞株SW-480增殖的影响，并探讨其可能机制。 方法 不同浓度AGE干预SW-480细胞，噻唑蓝（MTT）法比较各组细胞活力，流式细胞术观察AGE对SW-480细胞周期的影响，蛋白质印迹法观察AGE对SW-480细胞CyclinD1表达的影响，端粒重复序列扩增法（telomeric repeat amplification protocol，TRAP）银染法观察AGE对SW-480细胞端粒酶活性的影响。MTT测细胞活力的检测设置空白对照组、100 μg/mL小牛血清白蛋白（bovine serum albumin，BSA）组及50、100、500 μg/mL AGE组，其余检测只设置100 μg/mL BSA组和100 μg/mL AGE组。 结果 MTT结果示AGE促进SW-480细胞的增殖，且呈浓度依赖性。100 μg/mL BSA组与100 μg/mL AGE组72 h后的细胞G0/G1期所占百分比分别为56.02%±0.58%、51.93%±1.01%，差异有统计学意义（Plt;0.05）。蛋白质印迹法示100 μg/mL AGE组72 h后CyclinD1的表达较100 μg/mL BSA组增加，差异有统计学意义（Plt;0.05）。TRAP银染法检测示100 μg/mL AGE干预SW-480细胞72 h后可以增加端粒酶活性（Plt;0.05）。 结论 AGE可促进人结肠癌细胞SW-480生长，呈剂量依赖性。其作用机制可能与AGE上调CyclinD1的表达加速G1/S期转换及增加端粒酶活性有关。【Abstract】 Objective To observe the effects of advanced glycosylation end products (AGE) on proliferation of SW-480 cells and study the possible mechanism. Methods Various concentrations of AGE were designed to have impact on SW-480 cells. Proliferation of SW-480 cells was assessed by thiazolyl blue tetrazolium bromide (MTT) assay; The impact of AGE on the cell cycle of SW-480 cells was analyzed by flow cytometry (FCM); the influence of AGE on expression of CyclinD1 was checked by Western blotting; and the impact of AGE on telomerase activity was examined by telomeric repeat amplification proctol (TRAP) sliver staining. For the MTT assay, blank control group, 100 μg/mL bovine serum albumin (BSA) group, 50, 100 and 500 μg/mL AGE groups were designed, while for other examinations, there were only 100 μg/mL BSA group and 100 μg/mL AGE group. Results MTT result showed that AGE increased the proliferation of SW-480 cells in a dose-dependent mode. The proportion of the cells at G0/G1 stage of the 100 μg/mL BSA group and the 100 μg/mL AGE experimental group were (56.02±0.58)% and (51.93±1.01)% respectively after 72 hours, with a significant difference (Plt;0.05); western blotting showed that the expression of CyclinD1 in the 100 μg/mL AGE group was significantly higher than that in the 100 μg/mL BSA group after 72 hours; TRAP silver staining demonstrated that telomerase activity increased significantly after treated with 100 μg/mL AGE for 72 hours. Conclusions AGE can promote the growth of SW-480 cells in a dose-dependent mode. Its mechanism is mainly by up-regulating the expression of CyclinD1 to shorten G0/G1 and increasing the telomerase activity significantly.