ObjectiveTo observe and analyze the effect of HbA1c level on macular microcirculation in patients with type 2 diabetes mellitus (T2DM).MethodsA cross-sectional study. One hundred and twenty-four T2DM patients (124 eyes) without diabetic retinopathy who diagnosed by the examination of fundus color photography in Lixiang Eye Hospital Of Soochow University during September to December 2017 were enrolled in this study. There were 59 males (59 eyes) and 65 females (65 eyes), with the mean age of 65.06±7.99 years old. All patients underwent BCVA, fundus color photography, and OCT angiography (OCTA). The history of diabetes, hypertension and dyslipidemia were recorded in detail. According to the HbA1c level, patients were divided into three groups, HbA1c ideal control group (group A, HbA1c ＜7%, 67 eyes), HbA1c control group (group B, 7%≤HbA1c≤9%, 44 eyes), and HbA1c poor control group (group C, HbA1c＞9%, 13 eyes), respectively. The 3 mm×3 mm range of the macular area was scanned by OCTA instrument. The vascular density (VD) and skeleton density (SD) of nonsegmented retinal layer (NRL), superficial retinal layer (SRL) and deep retinal layer (DRL) in the macular area and foveal avascular zone (FAZ) area, non-circularity index, axial rate (AR) of SRL were measured. The correlation between HbA1c, BCVA and VD, SD of NRL, SRL, DRL was analyzed statistically with Spearman correlation test. The correlation between systemic factors and the above indicators was analyzed statistically with linear regression analysis.ResultsThe results of linear regression analysis showed that HbA1c was significantly correlated with VD (t=?3.237, ?3.156, ?2.050) and SD (t=?0.3.45, ?3.034, ?2.248) of NRL, SRL and DRL (P＜0.05); but no correlation with FAZ, non-circularity index and AR (t=1.739, 0.429, 1.155; P＞0.05). The differences of VD (F=6.349, 5.981, 3.709), SD (F=7.275, 6.085, 1.904) and AR (F=0.027) of NRL, SRL and DRL in group A, B and C were statistically significant (P＜0.05); but the differences of FAZ (F=1.904), non-circularity index (F=0.280) was not statistically significant (P＞0.05). Significant differences (P＜0.05) of VD and SD of NRL were found between group A and B (t=1.987, 2.201), group A and C (t=3.365, 3.572), group B and C (t=2.010, 2.076). Significant differences (P＜0.05) of VD and SD of SRL were found between group A and B (t=2.087, 2.168), group A and C (t=3.197, 3.194). There were significant differences (P＜0.05) in SD of DRL between group A and B (t=2.239), group A and C (t=?2.519). There was significant difference in VD of DRL between group A and C (t=2.363). The results of Spearman correlation analysis showed that HbA1c was negatively correlated with VD (r=?0.273, ?0.255, ?0.222; P=0.002, 0.004, 0.013) and SD (r=?0.275, ?0.236, ?0.254; P＜0.05) of NRL, SRL, DRL; positively correlated with FAZ and BCVA (r=0.221, 0.183; P＜0.05). BCVA was negatively correlated with VD (r=?0.210, ?0.190, ?0.245) and SD (r=?0.239, ?0.207, ?0.296) of NRL, SRL, and DRL (P＜0.05), but not correlated with FAZ (r=0.099, P＞0.05).ConclusionThe decrease of macular perfusion and the morphological change of FAZ accompanied by HbA1c increased.
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)
Objective
To study the effects of advanced glycation end (AGEs) products induced by bovine serum albumin (BSA) on the survival and the morphology of bovine retinal endothelial cells (BREC) and pericytes (BRP).
Methods
BSA with the final concentration of 50 mg/ml was incubated in PBS, containing 500 mmol/L D-glucose, for 12 weeks under 37℃. AGEs-BSA was purified by Sephacryl S-300 column chromatography and was confirmed by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The concentration of AGEs-BSA was determined by the method of commassie protein assay. In order to detect the toxic effects of AGEs-BSA on cultured BREC and BRP, groups of AGEs-BSA and BSA with different concentration and untreated control were set up. Phase contrast microscope was used to observe the effect of AGEs-BSA and BSA (with the concentration of 500mu;g/ml and actuation duration of 48 hours) on morphology of BREC and BRP.
Results
As the dosage of AGEs-BSA increased, the number of inhibited cells increased. When the concentration of AGEs-BSA was 500mu;g/ml, the inhibited BREC in AGEs-BSA group was (72.8plusmn;15.9)% of which in untreated control group, and the inhibited BRP was (64.8plusmn;9) % of which in untreated control group. AGEs-BSA with low concentration promoted the proliferation of endothelial cells, but there was no significant difference between AGEs-BSA and the control group (P=0.231). Inhibited proliferation and abnormal morphology were seen under the phase contrast microscope while the normal morphology of cells was found in BSA and control group.
Conclusion
AGEs-BSA with the high concentration may inhibit the growth of both BREC and BRP, which leads the loss of BRP and damage of vascular function. These results suggest that nonenzymatic glycosylation plays a major role in diabetic complications.
(Chin J Ocul Fundus Dis, 2006, 22: 11-15)
【摘要】 目的 觀察晚期糖基化終產物（advanced glycosylation end prodrcts，AGE）對人結腸癌細胞株SW-480增殖的影響，并探討其可能機制。 方法 不同濃度AGE干預SW-480細胞，噻唑藍（MTT）法比較各組細胞活力，流式細胞術觀察AGE對SW-480細胞周期的影響，蛋白質印跡法觀察AGE對SW-480細胞CyclinD1表達的影響，端粒重復序列擴增法（telomeric repeat amplification protocol，TRAP）銀染法觀察AGE對SW-480細胞端粒酶活性的影響。MTT測細胞活力的檢測設置空白對照組、100 μg/mL小牛血清白蛋白（bovine serum albumin，BSA）組及50、100、500 μg/mL AGE組，其余檢測只設置100 μg/mL BSA組和100 μg/mL AGE組。 結果 MTT結果示AGE促進SW-480細胞的增殖，且呈濃度依賴性。100 μg/mL BSA組與100 μg/mL AGE組72 h后的細胞G0/G1期所占百分比分別為56.02%±0.58%、51.93%±1.01%，差異有統計學意義（Plt;0.05）。蛋白質印跡法示100 μg/mL AGE組72 h后CyclinD1的表達較100 μg/mL BSA組增加，差異有統計學意義（Plt;0.05）。TRAP銀染法檢測示100 μg/mL AGE干預SW-480細胞72 h后可以增加端粒酶活性（Plt;0.05）。 結論 AGE可促進人結腸癌細胞SW-480生長，呈劑量依賴性。其作用機制可能與AGE上調CyclinD1的表達加速G1/S期轉換及增加端粒酶活性有關。【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.
ObjectiveTo summarize the characteristics of hexosamine biosynthesis pathway (HBP) and O-glycosylation and their roles in metabolism of hepatocellular carcinoma. MethodTo review the current literatures on the role of HBP and O-glycosylation in tumors, especially in hepatocellular carcinoma. ResultsThere was metabolic reprogramming in hepatocellular carcinoma cells, and the HBP was a branch of glycolysis pathway, which played an important role in tumorigenesis, development, and metastasis. HBP provided uridine diphosphate-N-acetylglucosamine (UDP-GlcNAc) for O-glycosylation, UDP-GlcNAc was a substrate for OGT, participating in O-glycosylation. O-glycosylation was a type of posttranslational modification that regulates the biological behavior of tumor cells by glycosylation of target proteins in tumor cells. ConclusionHBP and O-glycosylation can be used as intervention targets in the treatment of hepatocellular carcinoma, which provides a potential method for scientific prevention and treatment of hepatocellular carcinoma.
Objective To explore the influence and mechanism of mechanistic target of rapamycin kinase (mTOR)/ receptor of advanced glycation end products (RAGE) pathway mediated-ferritinophagy on high glucose consumption promoting invasion and migration of colorectal cancer (CRC). Methods① Patients and tissue samples. Clinical data and tissues were collected from CRC patients underwent surgery and completed the dietary questionnaire in the Second Affiliated Hospital of Harbin Medical University from October 2022 to October 2023. Real-time quantitative reverse transcription PCR (qRT-PCR) was used to analyzed the expression of nuclear receptor coactivator 4 (NCOA4) and ferritin in CRC and para-carcinoma tissues. ② Cell culture and treatment. The HT29 and HCT116 cells were treated by RPMI1640 medium containing 0, 35, 70, 105, 140 mmol/L glucose, and cell counting kit-8 (CCK-8) and lactate dehydrogenase (LDH) activity analysis were performed to confirm 105 mmol/L glucose was the optimal concentration in the current study. Then the HT29 and HCT116 cells were randomly divided into: control group, glucose group; control group, glucose group, si-RAGE group, and glucose+si-RAGE group; control group, glucose group, rapamycin group, and glucose+rapamycin group. Untreated HT29 and HCT116 cells were considered as control group. The cells in glucose group were treated with 105 mmol/L glucose for 48 h. The CRC cells in the si-RAGE group were transfected with si-RAGE for 6 h. The CRC cells in the rapamycin group were treated with 10 nmol/L rapamycin for 48 h. The CRC cells in the glucose+si-RAGE group were treated with 105 mmol/L glucose for 48 h combination transfected with si-RAGE for 6 h. The CRC cells in the glucose+rapamycin group were treated with 105 mmol/L glucose for 48 h combination treated with 10 nmol/L rapamycin for 48 h. Then electron microscopy and Western blot, wound healing assay and transwell assay were exhibited, respectively. ③ Azoxymethane (AOM)-induced CRC rat model. The effects of glucose consumption on malignant behavior and ferritinophagy mediated by mTOR/RAGE pathway were evaluated in AOM-induced CRC rat models. A total of 16 rats were randomly divided into control group and glucose group, the CRC number was recorded and HE staining of CRC tissues was further performed. The expression of RAGE, mTOR, NCOA4, and ferritin in colorectal tissues of rats from each group was detected by RT-qPCR. Results① More lymphatic node metastasis and TNM Ⅲ/Ⅳ stages were observed in CRC patients from high glucose consumption group (P=0.004, P=0.004). Moreover, we confirmed that NCOA4 expression was significantly decreased (P<0.001) while ferritin was significantly increased (P<0.001) in CRC tissues especially in the CRC tissues from patients with positive lymph nodes metastasis. ② High glucose treatment significantly decreased autophagosomes in HT29 and HCT116 cells while si-RAGE transfection increased autophagic vacuoles compared to the control group. When compared with the glucose group, autophagosomes were increased in the glucose+si-RAGE group. Moreover, compared to the control group, the expressions of RAGE, p-mTOR, and ferritin were increased (P<0.001) while the expression of NCOA4 was decreased (P<0.001) in glucose group, but the expressions of RAGE, p-mTOR, and ferritin were decreased (P<0.001) while the expression of NCOA4 was increased (P<0.001) in the si-RAGE group; when compared with the glucose group, the expressions of RAGE, p-mTOR, and ferritin were downregulated (P<0.001) while the expression of NCOA4 was upregulated (P<0.001) in HT29 and HCT116 cells from the glucose+si-RAGE group. Compared to the control group, the HT29 and HCT116 cells in the glucose group performed enhanced wound scratch healing , migration and invasion viabilities (P<0.05); but the HT29 and HCT116 cells in the si-RAGE group performed impaired wound scratch healing, migration and invasion viabilities (P<0.05). When compared with the glucose group, the HT29 and HCT116 cells in the glucose+si-RAGE group performed impaired wound scratch healing, migration and invasion viabilities (P<0.05). ③ Rapamycin treatment significantly inhibited the expression of ferritin (P<0.05) but induced the expression of NCOA4 (P<0.05) compared to the control group. When compared with the glucose group, the expression of ferritin was downregulated (P<0.05) while the expression of NCOA4 was upregulated (P<0.05) in HT29 and HCT116 cells from the glucose+rapamycin group. Additionally, compared to the control group, rapamycin treatment performed inhibited effect on wound scratch healing, migration and invasion viabilities in the HT29 and HCT116 cells (P<0.05); while the HT29 and HCT116 cells in the glucose+rapamycin group performed impaired wound scratch healing, migration and invasion viabilities (P<0.05) when compared with the glucose group. ④ In the AOM induced CRC rat models, we found the more CRCs, aggravated cellular pleomorphism and upregulated expressions of RAGE, mTOR, ferritin (P<0.05) while downregulated expression of NCOA4 (P<0.05) in the glucose group than those of the control group. ConclusionHigh glucose consumption promote invasion and migration in CRC through suppressing ferritinophagy via activating the mTOR/RAGE pathway.
