ObjectiveTo investigate the expression and mechanism of miR-1470 in plasma of diabetic retinopathy (DR) patients.MethodsThirty patients with DR (DR group), 30 patients with diabetes (DM group) and 30 normal healthy subjects (normal group) were enrolled in the study. Three groups of subjects were taken 5 ml of venous blood, and total plasma RNA was extracted and purified. The differentially expressed miRNAs in the plasma of DR patients were screened by gene chip, and the results of gene chip detection were verified by reverse transcription polymerase chain reaction (RT-PCR). Bioinformatics was used to predict potential target genes for miRNA regulation, and miR-1470 and its target gene epidermal growth factor receptor (EGFR) were screened. Human retinal microvascular endothelial cells (hREC) were divided into normal group (sugar concentration 5.5 mmol/L) and high glucose group (sugar concentration 25.0 mmol/L). hREC was transfected into miR-1470 mimics to establish a miR-1470 high expression cell model, which was divided into blank control group, high expression group and negative control group. The expression of miR-1470 was detected by RT-PCR. The expression of EGFR protein was detected by Western blot. The measurement data of the two groups were compared using the independent sample t test. The comparison of the measurement data between the two groups was analyzed by ANOVA. The comparison between the measurement data of the groups was compared by multiple comparisons.ResultsThe results of RT-PCR were consistent with those of the gene chip. The expression of miR-1470 in the plasma of the DR group, the DM group and the normal group was statistically significant (F=63.486, P=0.049). Compared with the DM group and the normal group, the expression of miR-1470 in the DR group was significantly decreased, and the difference was statistically significant (q=111.2, 73.9; P＜0.05). The expression of miR-1470 in hREC in the high glucose group was significantly lower than that in the normal group (t=42.082, P=0.015). The expression of EGFR protein in hREC of high glucose group was significantly higher than that of normal group (t=−39.939, P=0.016). The expression of miR-1470 (F=637.069, P=0.000) and EGFR (F=122.908, P=0.000) protein expression in hREC of blank control group, negative control group and high expression group were statistically significant . Compared with the blank control group and the negative control group, the expression of miR-1470 in hREC of high expression group was significantly increased (q=329.7, 328.8; P＜0.05), and the expression of EGFR protein was significantly decreased (q=242.5, 234.6; P＜0.05). There was no significant difference in the expression of miR-1470 and EGFR protein in hREC between the negative control group and the blank control group (q=1.5, 7.9; P＞0.05).ConclusionThe expression of miR-1470 in the plasma of patients with DR is significantly down-regulated, and the increase of EGFR expression may be related to it.
ObjectiveTo observe the effects of four prostaglandin E2 (PGE2) receptors (EP1-4R) on the activation of inflammasomes and cell damage in human retinal microvascular endothelial cells (hRMEC) in a high glucose environment.MethodsThe hRMEC were divided into normal group and high glucose group, and they were cultured in Dulbecco modified Eagle medium containing 5.5 and 30.0 mmol/L glucose, respectively. Flow cytometry was used to observe the apoptosis rate of the high glucose group and the normal group; enzyme chain immunosorbent assay (ELISA) was used to detect the level of PGE2 in the culture supernatant of hRMEC cells. Western blot was used to detect the protein expression of cyclooxyganese (COX2) and EP1-4R in hRMEC. Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of EP1-4R mRNA in hRMEC. After 72 h of culture, the cells in the high glucose group were divided into control group, PGE2 group, EP1-4R agonist group, PGE2+EP1-4R inhibitor group, and dimethylsulfoxide group. According to the group, each group was given the corresponding agonist or inhibitor to continue the culture for 24 h. QRT-PCR was used to detect the expression of nucleotide-binding oligomerization structure-like receptor protein (NLRP3) and pro-interleukin (IL)-1β mRNA in each group of cells. ELISA was used to detect the content of IL-1β and lactic dehydrogenase (LDH) in the cell culture supernatant. Western blot was used to detect the expression of cleaved Caspase-1 in each group of cells. At the same time, hRMEC in a high glucose environment was given IL-1β stimulation for 24 h, and the activity of LDH in the supernatant of the cell culture medium was detected.ResultsThe apoptotic rate, COX2 protein expression, and PGE2 protein content in hRMEC in the high glucose group were significantly higher than those in the normal group, and they were time-dependent. Compared with the normal group, the expression levels of EP1R, EP2R, EP4R protein and mRNA in hRMEC in the high glucose group were higher than those in the normal group (P＜0.05). Compared with the control group, PGE2 group (t=4.627, P＜0.01), EP1-4R agonist group (t=3.889, 3.583, 2.445, 3.216; P＜0.05) hRMEC NLRP3 mRNA expression level was significantly increased; the expression level of pro-IL-1β mRNA increased, however the difference was not statistically significant (PGE2 group: t=1.807, P＞0.05; EP1-4R agonist group: t=1.807, 1.477, 0.302, 1.926, P＞0.05). Compared with the PGE2 group, the expression of NLRP3 mRNA in hRMEC in the PGE2+EP2R inhibitor group was significantly reduced (t=2.812, P＜0.05); the expression of pro-IL-1β mRNA in hRMEC in the PGE2+EP3R inhibitor group was significantly increased (t=4.113, P＜0.01). The protein content of IL-1β in the cell culture supernatant of the PGE2 group, EP1R agonist group and EP2R agonist group was significantly higher than that of the control group (t=5.155, 4.136, 4.817; P＜0.01). Compared with PGE2 group, the protein content of IL-1β in the cell culture supernatant of the PGE2+EP2R inhibitor group and the PGE2+EP4R inhibitor group were significantly lower than that of the PGE2 group (t=1.964, 4.765; P＜0.05). The expression of cleaved Caspase-1 in hRMEC in the PGE2 group and EP2R agonist group was significantly higher than that in the control group (t=5.332, 4.889; P＜0.05). The expression of cleaved Caspase-1 in hRMEC in the PGE2+EP2R inhibitor group was significantly lower than that of the PGE2 group (t=6.699, P＜0.01). The LDH activity in the cell culture supernatant of the PGE2 group and the EP2R agonist group was significantly higher than that of the control group (t=4.908, 4.225; P＜0.05). The activity of LDH in the cell culture supernatant of the PGE2+EP2R inhibitor group was significantly lower than that of the PGE2 group (t=5.301, P＜0.01). Compared with the control group, the LDH activity in the culture supernatant of hRMEC cells in the high glucose environment was significantly increased (t=3.499, P＜0.05).ConclusionsThe four receptors of PGE2 can activate NLRP3 and its effector molecules to varying degrees. EP2R mainly mediates hRMEC damage under high glucose environment.
Objective To estimate the prevalence of diabetic retinopathy (DR) in residents at the age of ge; 60 years in Beixinjing blocks, Shanghai, and to analyze the changes and risk factors of the occurrence of DR while comparing with the previous epidemiology study in the same blocks in 2003.Methods With the help of an established resident health data,a epidemiology study was performed on all of the old residents (ge; 60 years old) with diabetes mellitus from November 2007 to April 2008.The epidemiology data included resident common information, health check, examination of visual acuity,slitlamp biomicroscopy, direct ophthalmoscopy, and photography of 2 pictures covering 450 posterior ocular fundus area acquired by digital nonmydriasis fundus camera. The diagnosis of DR was drawn according to the international DR classification (2002). The DR grade was compared in 254 residents who were involved in both of the two epidemiology studies in 2003 and 2007.Chisquare method was used to analyze the risk factors of DR and stepwise logistic analysis was used for identifying the independent factors.Results A total of 483 diabetic residents was included in this study with the inclusion rate of 91.30%, in whom 121 DR patients was found with the DR prevalence of 25.05％. The prevalence of non-proliferative DR and proliferative DR was 22.36% and 2.69%, respectively. The duration of diabetes was confirmed as the independent risk factor of DR prevalence, and no risk factor was found to affect the different ratio of non-proliferative and proliferative DR. Comparing to the results of the study in 2003,the DR grade remained steadily or decreased in 92.52% of the 254 diabetes residents.Conclusion The prevalence of DR is high in the diabetic residents at the age of ge; 60 years.The intervention work of DR prevention in Beixinjing blocks is effective.
Objective To investigate the prevalence of high myopia，the prevalence and risk factors of high myopia associated with chorioretinopathy in residents aged 60 years or over in Beixinjing community, Shanghai, China.Methods A cluster stratified random sampling method was used to screen 4153 people aged 60 and over in Beixinjing community. There were 3851participants in total with a 9273% response rate. Participants were invited to complete a questionnaire and received a comprehensive eye examination including visual acuity, refraction, slitlamp microscopic examination, direct ophthalmoscopy and fundus photography and so on. Spherical equivalent (SE) was used to determine the degree of refractive errors. The diagnosis of high myopic chorioretinopathy was made if SEgt;-6.00 D and myopic chorioretinal atrophy lesions were presented such as posterior scleral staphyloma, lacquer cracks, Fuchs spot and myopic arc spots. The degree of visual acuity impairment was determined according to the World Health Organization (WHO) classification as low vision (the best corrected visual acuity ge; 0.05, but lt;0.3) or blindness (the best corrected visual acuity lt;0.05).Results There were 207/3851(5.37%) high myopia patients, in which 183/207 (88.40%) patients were associated with myopic chorioretinopathy. The prevalence of myopic chorioretinopathy decreased while age increased (chi;2=19.21, Plt;0.01), but statistically there was no gender difference （chi;2=1.83, Pgt;0.05）. Logistic regression analysis showed that there were significant differences in the prevalence of high myopia between people with different age, educational levels and family history （chi;2=19.21，32.08，960.68；Plt;0.01）.There were 29 cases of bilateral blindness, 96 cases of unilateral blindness,104 cases of bilateral low vision and 562 cases of unilateral low vision in those participants. In 183 cases of high myopic chorioretinopathy patients, 111（60.65 %）cases had an obvious visual impairment, including 3448% (10 cases) of bilateral blindness, 1146% (11 cases) of unilateral blindness, 2981% (31 cases)of bilateral low vision and 1050% (59 cases) of unilateral low vision.Conclusions The prevalence of high myopia of residents aged ge;60 years in Beixinjing community, Shanghai,China is relatively high. Age, education level and family history are the most important factors affecting the occurrence of chorioretinopathy in high myopia patients.
ObjectiveTo observe the changes in open probability and protein expression of large conductance Ca2+-activated K+ (BK) channel in retinal vascular smooth muscle cells (RVSMCs) of diabetic rats. MethodsStreptozotocin (STZ)-induced rat diabetic animal model was established by STZ injection intraperitoneally.RVSMCs were isolated by enzyme digestion. The BK currents in control and diabetic groups were recorded by patch clamp technique in single channel configuration. BK channel protein expression in control and diabetic group were measured by Western blot. ResultsCompared with control group, the NP0 of BK channels in diabetic group were significantly increased (t=4.260, P < 0.05). Compared with control group, there was no significant difference inα-subunit protein expression in diabetic group in RVSMCs (t=10.126, P > 0.05); however, β1-subunit protein expression was remarkably increased in diabetic group (t=5.146, P < 0.05). ConclusionThe NP0 of BK channels andβ1-subunit protein expression are increased in RVSMCs of diabetic rats.
Objective To measure the concentration of serum transthyretin (TTR) of patients with different stages of diabetic retinopathy (DR). Methods A total of 176 patients with diabetes mellitus were included in this study. There were 104 males and 72 females. The patients aged from 21 to 74 years, with the mean age of (56±11) years. The diabetes duration raged from 1 to 30 years, with the mean diabetes duration of (10±7) years. The HbA1C was 5.2%−14.1%, with the mean HbA1C of (8.6±2.0)%. According to the fundus examination, 58 patients had DR (33.0%), but the other 118 patients not (67.0%). For these DR patients, 10 patients were in stage Ⅰ (5.7%), 26 patients in stage Ⅱ (14.8%), 8 patients in stage Ⅲ (4.5%), and 14 patients in stage Ⅳ (8.0%). The concentration of serum TTR was measured by enzyme-linked immunosorbentassay kit. The differences in the concentration of serum TTR between different DR stages were compared.Bivariate analysis was used to analyze the influencing factors of TTR. Results The concentrations of serum TTR of the patients without DR or with DR of stage Ⅰ to Ⅳ were (224.96±65.47), (383.68±102.99), (247.44±63.21), (228.2±45.89), (189.34±70.12) mg/L, respectively. The difference between different DR stages was statistically significant (F=14.690,P＜0.001).Bivariate analysis showed that the concentration of TTR was correlation to DR (r=0.179,P=0.017). There was no correlation between the concentration of TTR and diabetes duration (r=−0.027,P=0.727), hypertension (r=0.018,P=0.810), hyperlipoidemia (r=0.101,P=0.182), and the use of insulin (r=−0.032,P=0.675). Conclusion The concentration of serum TTR was increased in early DR patients, and gradually decreased with the progression of DR. The concentration of TTR is correlated to DR.
Objective To investigate the prevalence rate and risk factors of diabetic retinopathy (DR) in residents aged 50 and above in Binhu Area, Wuxi City. Methods Stratified cluster random sampling method was used to investigate the prevalence of DR of residents aged 50 and above from January to December 2010. Detailed medical history, general examination, visual acuity, slit lamp microscope, direct ophthalmoscopy and blood sample testing were conducted for all selected subjects. DR diagnostic criteria refer to the 2002 International DR classification criteria. Unilateral or bilateral DR was both considered as DR patients.χ2 test was used for statistical analysis of risk factors, and independent risk factors were screened by SPSS 17.0 software. Results A total of 6150 residents underwent the examination with a participating rate of 91.5%. Seven hundred and three residents were diagnosed as with diabetes, in which 40 were unable to record the fundus condition due to opaque refractive media. Among the rest 663 diabetic patients, 36 (5.4%) were DR patients. There were 34 non-proliferative DR (5.1%) and two proliferative DR (0.3%). The duration of diabetes and fasting blood-glucose were the independent risk factor of DR (r=0.008, 0.009; P＜0.05). Conclusions The prevalence rate of DR is low in the residents aged 50 and above in Binhu Area, Wuxi City. The duration of diabetes and fasting blood-glucose level were confirmed as the independent risk factor of DR.
Objective To investigate the causes of blindness and low vision of the people over 50 years old in Binhu Area of Wuxi City. Methods Cluster sampling was used in randomly selected individuals over 50 years old in 28 villages. The oculopathy related to blindness and low vision of the people over 50 years old were analyzed. The vision acuity lt;0.05 was defined as blindness, while 0.05-0.3 was defined as low vision. SPSS 17.0 software was used to analyze the data. Results Totally 6150 individuals were examined. The bilateral blindness and low vision was found in 47 and 84 individuals, unilateral blindness and low vision was found in 201 and 214 individuals. Among 201 unilateral blindness individuals, there were 55 individuals have low vision in the other eye. In the 295 blind eyes, there are 116 eyes (39.32%) with cataract, 31 eyes (10.51%) with high myopia macular degeneration, and 28 (9.49%) eyes with atrophy eyeballs. In the 437 low vision eyes, there are 223 eyes (51.03%) with cataract, 41 (9.38%) eyes with high myopia macular degeneration, and 41 (9.38%) eyes with age-related macular degeneration. Conclusion Cataract is the first cause leading to blindness and low vision, followed by age-related macular degeneration, high myopia macular degeneration and atrophy eyeballs.