Effect of leptin on the oxidative damage in human retinal pigment epithelial cells_Chinese Journal of Ocular Fundus Diseases

Authors：

LuoYunfeng , NiYan ,  LuanJie

Department of Ophthalmology, Zhongda Hospital, Southeast University, Nanjing 210009, China;

Corresponding author：

LuanJie, Email: luanqiu10@163.com

Keywords：

Retinal pigment epithelium/injuries; Leptin/poisoning; Reactive oxygen species/poisoning

DOI：

10.3760/cma.j.issn.1005-1015.2014.02.011

Video：

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Objective To investigate the effects of leptin on the oxidative damage in human retinal pigment epithelial (RPE) cells. Methods Human RPE cells (ARPE-19) were cultured in vitro, and randomly divided into control group and insulin resistance group. RPE cells were treated with 0, 10, 100 ng/mL leptin for 24, 48, 72 hours respectively. Then the levels of reactive oxygen species (ROS) expression in RPE cells were detected by 2', 7'-dichlorofluorescin-diacetate (DCFH-DA), and the levels of 8-hydroxy-2'-deoxyguanosine (8-OHdG) expression in RPE cells were observed by immunocytochemistry (ICC), and the levels of human 8-oxoguanine DNA glycosylase l (hOGG1) expression in lysate were measured by Western blot. Results After 24, 48, 72 hours, the level of ROS (Control group:F=37.136, 37.178, 49.634; P < 0.05. Insulin resistance group:F=9.822, 28.881, 71.150;P < 0.05), 8-OHdG (Control group:F=88.643, 390.920, 1039.276;P < 0.05.Insulin resistance group:F=273.311, 299.155, 82.237;P < 0.05) and hOGGl (Control group:F=470.062, 1073.113, 295.456;P < 0.05. Insulin resistance group:F=240.032, 592.389, 527.760;P < 0.05) expression increased significantly with the increase of leptin concentration in control group and insulin resistance group. Under the same leptin concentration, the level of 8-OHdG has a trend that it was higher in the insulin resistance group than the control group. After 24 hours, the difference of hOGGl expression between control group and insulin resistance group was not significant (F=23.392, P > 0.05). After 72 hours, the level of hOGGl expression was significantly higher in the insulin resistance group than the control group (F=129.394, P < 0.05). The level of hOGGl expression was significantly higher at 48 hours than that at 24 hours and 72 hours (P < 0.05). Conclusion Leptin could induce the oxidative damage of RPE cells in normal and insulin resistance status. With the increase of leptin concentration and time extended, the degree of oxidative damage and its repair were both increased. The degree of oxidative repair increased with the increase of leptin concentration, but decreased with time extended.

Citation： LuoYunfeng, NiYan, LuanJie. Effect of leptin on the oxidative damage in human retinal pigment epithelial cells. Chinese Journal of Ocular Fundus Diseases, 2014, 30(2): 161-165. doi: 10.3760/cma.j.issn.1005-1015.2014.02.011 Copy

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2. Nguyen TT, Wong TY.Retinal vascular manifestations of metabolic disorders[J]. Trends Endocrinol Metab, 2006, 17:262-268.
3. Jiao J, Li Y. C-peptide and diabetic retinopathy[J].Int J Ophthalmol, 2008, 8:1441-1443.
4. Mora M, Mansego ML, Serra-Prat M, et al.Glucose impairment and ghrelin gene variants are associated to cognitive dysfunction[J].Aging Clin Exp Res, 2014, 26:161-169.
5. 傅敏, 栾洁.可见光对视网膜色素上皮细胞氧化损伤的影响.中华眼底病杂志, 2011, 27:466-470.
6. Brownlee M.Biochemistry and molecular cell biology of diabetic complications[J].Nature, 2001, 414:813-820.
7. Gariano RF, Nath AK, D'Amico DJ, et al.Elevation of vitreous leptin in diabetic retinopathy and retinal detachment[J].Invest Ophthalmol Vis Sci, 2000, 41:3576-3581.
8. Maberley D, Cui JZ, Matsubara JA.Vitreous leptin levels in retinal disease[J].Eye (Lond), 2006, 20:801-804.
9. Cheng Y, Ren X, Gowda AS, et al.Interaction of Sirt3 with OGG1 contributes to repair of mitochondrial DNA and protects from apoptotic cell deathunder oxidative stress[J/OL].Cell Death Dis, 2013, 4:E731[2013-07-18].http://www.nature.com/cddis/journal/v4/n7/full/cddis2013254a.html.

Chinese Journal of Ocular Fundus Diseases

Effect of leptin on the oxidative damage in human retinal pigment epithelial cells

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