1.Visible light induce oxidative damage to human retinal pigment epithelial cells
Min FU ; Jie LUAN ; Zhanghong ZHANG
Chinese Journal of Ocular Fundus Diseases 2011;27(5):466-470
ObjectiveTo observe the effect of visible light (white light, red light, blue light) on the expression of reactive oxygen species (ROS), 8-OHdG and hOGG1 in cultured human retinal pigment epithelial (RPE) cells. MethodsCultured human RPE-19 cells (4th-6th generations) were divided into white light,red light, blue light and control group.The illumination was 600 Lux.The cells of experimental groups were exposed to white light or red light for 6, 12, 24 and 48 hours, and exposed to blue light for 1, 3, 6 and 12 hours, while cells of the control group were cultured in foil packaged dishes to avoid light. The levels of ROS expression were detected by 2′, 7′-dichlorofluorescin-diacetate (DCFH-DA), the levels of 8-OHdG protein expression were observed by immunocytochemistry (ICC), and the levels of hOGG1 were measured by western blot. ResultsCompared to the control group, the ROS expression in RPE cells were increased in white and red light group after 12, 24 and 48 hours and in blue light group after 1, 3, 6 and 12 hours (Fwhite light=11. 611, Fred light =6.706, Fblue light =23. 259; P<0.05 ). Additionally, the ROS expression had a tendency to increase gradually along with exposure time. Compared to the control group, the 8-OHdG expression in RPE cells were increased significantly in both white and red light group after 12, 24 and 48 hours and in blue light group after 1, 3, 6 and 12 hours (Fwhite light =16. 032,Fred light =6. 378, Fblue light =19. 484; P<0.05). Additionally, the 8-OHdG expression in white and red light group were increased gradually with exposure time but decreased when exposure time was up to 48 hours, while that in blue light group was increased firstly though it started to decrease when exposure time was up to 6hours. Compared to the control group, the hOGG1 expression in RPE cells were increased in white and red light group after 12, 24 and 48 hours and in blue light group after 6 and 12 hours (Fwhite light =15. 121,Fred light=21. 041,Fblue light12. 479; P<0.05). ConclusionsExposure to white, red or blue light could induce ROS production and DNA oxidative damage in RPE cells in a time-dependent way.Exposure to visible light could switch on self protection of RPE cells against DNA oxidative damage by up-regulating of the hOGG1 expression.
2.Effects of methylprednislone on alveola septal apoptosis in autoimmune emphysema rats
Jing HAN ; Chaojie ZHANG ; Cheng ZHANG ; Ping LU ; Zhanghong OUYANG ; Xiangyan ZHANG
Chinese Journal of Biochemical Pharmaceutics 2014;37(4):36-39
Objective To investigate the effects of methyprednislone on the expression of tumor alveolar septal cell apoptosis in autoimmune emphysema rats,in order to provide a theoretical basis for Chronic obstructive pulmonary disease(COPD)pathogenesis and it’s treatment.Methods 24 rats were randomly divided into three groups:normal control group (n=8 ),model group (n=8 )and intervention group (methylprednisolone sodium succinate,n=8).Intraperitoneal injection of primary cultured human umbilical vein endothelial cells were given to establish the autoimmune emphysema models,intervention group was injected with methylprednisolone at the meantime,and normal control group was received adjuvant only.Pathological changes were observed in lung tissues stained by hematoxylin eosin,mean liner intercept(MLI)and mean alveolar numbers(MAN)were measured.The localization of VEGF and VEGFR-2 in lung tissues were detected by immunohistochemical analysis.Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)technique was carried out to detect the alveolar septal cells apoptosis. Results The MLI in model group was higher than that in normal control group,while MAN was lower(P<0.05 );MLI in intervention group was lower than that in model group,but the MAN was higher (P<0.05 ).The localization of VEGF and VEGFR-2 in lung tissues in model group were lower than that in normal control group,and those in intervention group were higher than that in model group,the difference were all statistically significant(P<0.05 ).AI of alveolar septal cell in model group was higher than that in normal control group,which in intervention group was higher than that in model group,the difference were all statistically significant(P<0.05).Conclusion The decreasing of VEGF and VEGFR-2 in lung tissues may cause alveolar septal cell apoptosis and contribute to the pathogenesis of autoimmune emphysema of rats.Methyprednislone can alleviate the form of autoimmune emphysema in rats,which may be ralated to the regulation of VEGF and VEGFR-2 expression and inhibition of alveolar septal cell apoptosis.
3.The changes of miR-126 and SPRED1 levels after transient ischemic attack and their effects on prognosis
Lidong DING ; Zhanghong XIAO ; Tingting ZHANG ; Huawu MAO ; Xiaofang HANG ; Xiaobo LU ; Qi FANG
Chinese Journal of Geriatrics 2017;36(8):858-863
Objective To investigate the effects of changes of miR-126 and spouty related EVH,domain containing proteinl(SPRED1) after transient ischemic attack(TIA)on prognostic value for pathogenesis of secondary cerebral infarction.Methods Retrospective analysis of the clinical data of 106 patients with TIA was performed.The expression levels of miR-126,SPRED1 and vascular endothelial growth factor(VEGF)in peripheral blood were detected at 3 h,6 h and 12 h after TIA onset respectively.The specificity and sensitivity of miR 126 and SPRED1 in the diagnosis of TIA were analyzed.The miR-126 and SPRED1 levels versus ABCD2 score were compared for evaluating their predictive value in the diagnosis of secondary cerebral infarction within 30 days after TIA onset.Results The miR-126 level was declined after TIA onset at 3 h(9.41±1.04),especially at 12 h(6.59 ±2.78),versus in healthy control (9.35±1.76)(t =-7.764,P=0.000).The SPRED1 level after TIA onset was increased at 3 h(58.05 ± 17.53)pg/L,12 h(82.64 ± 18.60)pg/L versus in healthy control(52.38 ± 13.24)pg/L(t=12.374,P =0.000).A closely negative correlation was found between levels of miR 126 and SPRED1 at 12 h point but not at 3 h and 6 h(r=-0.278,P=0.004).Both miR-126 and SPRED1 levels at 12 h after TIA were implied to sensitivity and specificity evaluation.Additionally,VEGF was significantly increased at 3 h (345.61 ± 76.76) pg/L,6 h (461.65 ±103.87)pg/L and 12 h (519.22 ± 103.55)pg/L after TIA onset as compared with healthy control (107.77± 26.04) pg/L(t =26.569,29.756,34.699,all P =0.000).The decrease of miR-126 and increase of SPRED1 at 12h after TIA indicated high incidences of cerebral infarction but their significance was less than ABCD2 score.Combination of miR 126,SPRED1 and ABCD2 score significantly improved the prediction for cerebral infarction(Z=2.105,P =0.035).Conclusions After the onset of TIA,levels of miR-126 and SPRED1 expression in combination of ABCD2 score can improve predictive value for cerebral infarction development.