The effects of Curcumin on the cellular apoptosis induced by high glucose in rat retinal vascular endothelial cells_Chinese Journal of Ocular Fundus Diseases

Authors：

Huang Jiang , Li Yi , Xiao Jianjiang , Xu Guoxu , Bu Shuyang ,  Luo Weifeng

Department of Ophthalmology (Huang Jiang, Xu Guoxu,Bu Shuyang), Department of Neurology (Luo Weifeng), The Second Affiliated Hospital of Soochow University, Suzhou 215004,China; Department of Ophthalmology, The Huashan Hospital Affiliated of Fudan University, Shanghai 201907,China (Li Yi, Xiao Jianjiang);

Corresponding author：

Luo Weifeng, Email: lwfwxx@126.com

Keywords：

Curcumin; Retinal Vessels/pathology; Endothelial cells; Apoptosis; NF-κB

DOI：

10.3760/cma.j.issn.1005-1015.2017.05.017

Video：

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ObjectiveTo observe the effects of Curcumin on the cellular apoptosis of rat retinal vascular endothelial cells (RRVEC) induced by high glucose.MethodsGeneration 4 cultured RRVEC were used in this experiment, and identified with anti-vWF factor antibody by immunochemistry and immunofluorescence. The RRVEC were divided into control group (5.5 mmol/L glucose), high glucose group (30 mmol/L glucose), and treatment group (30 mmol/L glucose+30 μmol/L Curcumin), respectively. Flow cytometry was used to measure the cellular reactive oxygen species (ROS) level and apoptosis. The expression intensity and location of nuclear factor (NF)-κB p65 in the cells of the three groups were detected by immunochemistory. The expression of Bcl-2 and Bax protein was detected by Western blot test.ResultsImmunostaining showed that RRVEC were positive for vWF factor. The flow cytometry showed that the cellular ROS level in treatment group was higher than that in the control group (t=8.677, P=0.000), but less than that in the high glucose group (t=40.957, P=0.000). Compared with the high glucose group, the cellular ROS level in the treatment group was decreased significantly (t=6.568, P=0.000). The cellular apoptosis were significantly different among the three groups (F=325.137, P=0.000). Compared with the high glucose group, the cellular apoptosis in the treatment group was decreased significantly (t=12.818, P=0.000). Immunochemistry showed that NF-κB p65 was expressed strongly in the cellular nuclei and cytoplasm in the high glucose group than that in the control group and the treatment group with the significant differences (t=8.322, P=0.000). Western blot results demonstrated that compared with the control group, the expression of Bcl-2 of RRVEC and Bcl-2/Bax ratio decreased (t=4.362, 6.449; P=0.005, 0.001) and Bax increased (t=3.813, P=0.009）in the high glucose group, with statistically significant differences. Compared with the high glucose group, the expression of NF-κB and Bax decreased (t=2.577, 3.059; P=0.042, 0.022) and Bcl-2/Bax ratio increased significantly (t=3.831, P=0.009) in the treatment group.ConclusionCurcumin could suppress the cellular apoptosis of RRVEC induced by high glucose. The mechanism of Curcumin protecting RRVEC may be via regulating NF-κB signal pathway.

Citation： Huang Jiang, Li Yi, Xiao Jianjiang, Xu Guoxu, Bu Shuyang, Luo Weifeng. The effects of Curcumin on the cellular apoptosis induced by high glucose in rat retinal vascular endothelial cells. Chinese Journal of Ocular Fundus Diseases, 2017, 33(5): 513-517. doi: 10.3760/cma.j.issn.1005-1015.2017.05.017 Copy

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1. Kowluru RA, Kowluru A, Mishra M, et al. Oxidative stress and epigenetic modifications in the pathogenesis of diabetic retinopathy[J]. Prog Retin Eye Res, 2015, 48: 40-61. DOI:0.1016/j.preteyeres.2015.05.001.
2. Giordano CR, Roberts R, Krentz KA, et al. Catalase therapy corrects oxidative stress-induced pathophysiology in incipient diabetic retinopathycatalase therapy in incipient diabetic retinopathy[J]. Invest Ophth Vis Sci, 2015, 56(5): 3095-3102. DOI:10.1167/iovs.14-16194.
3. Behl T, Kaur I, Kotwani A. Implication of oxidative stress in progression of diabetic retinopathy[J]. Surv Ophthalmol, 2016, 61(2): 187-196. DOI:10.1016/j.survophthal.2015.06.001.
4. Bharti AC, Aggarwal BB. Nuclear factor-kappa B and cancer: its role in prevention and therapy[J]. Biochem Pharmacol, 2002, 64(5): 883-888. DOI: 10.1016/S0006-2952(02)01154-1.
5. Baeuerle PA, Baltimore D. NF-κB: ten years after[J]. Cell, 1996, 87(1): 13-20.DOI: 10.1016/S0092-8674(00)81318-5.
6. De Simone V, Franze E, Ronchetti G, et al. Th17-type cytokines, IL-6 and TNF-α synergistically activate STAT3 and NF-κB to promote colorectal cancer cell growth[J]. Oncogene, 2015, 34(27): 3493-3503.DOI:10.1038/onc.2014.286.
7. Gong G, Waris G, Tanveer R, et al. Human hepatitis C virus NS5A protein alters intracellular calcium levels, induces oxidative stress, and activates STAT-3 and NF-κB[J]. Proc Natl Acad Sci USA, 2001, 98(17): 9599-9604. DOI:10.1073/pnas.171311298.
8. Noorafshan A, Ashkani-Esfahani S. A review of therapeutic effects of curcumin[J]. Curr Pharm Design, 2013, 19(11): 2032-2046. DOI:10.2174/138161213805289273.
9. Trujillo J, Chirino YI, Molina-Jijón E, et al. Renoprotective effect of the antioxidant curcumin: recent findings[J]. Redox Bio, 2013, 1(1): 448-456.DOI:10.1016/j.redox.2013.09.003.
10. Hu S, Maiti P, Ma Q, et al. Clinical development of curcumin in neurodegenerative disease[J]. Expert Rev Neurother, 2015, 15(6): 629-637.DOI:10.1586/14737175.2015.1044981.
11. Hassaninasab A, Hashimoto Y, Tomita-Yokotani K, et al. Discovery of the curcumin metabolic pathway involving a unique enzyme in an intestinal microorganism[J]. Proc Natl Acad Sci USA, 2011, 108(16): 6615-6620.DOI: 10.1073/pnas.1016217108.
12. Oltval ZN, Milliman CL, Korsmeyer SJ. Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programed cell death[J]. Cell, 1993, 74(4): 609-619. DOI: 10.1016/0092-8674(93)90509-0.
13. 刘丽娅, 马景学, 刘丹岩, 等. 姜黄素对 IL-1β诱导的兔RPE细胞中核因子-κB 相关炎性因子表达的抑制作用[J].中华实验眼科杂志, 2016, 34(9): 804-812. DOI: 10.3760/cma.j.issn.2095-0160.2016.09.008.Liu LY, Ma JX, Liu DY, et al. Inhibition of curcumin on the expression of IL-1 β-induced nuclear factor-κB-dependent inflammatory gene in rabbit RPE cells[J].Chin J Exp Ophthalmol, 2016, 34(9): 804-812. DOI: 10.3760/cma.j.issn.2095-0160.2016.09.008.
14. Yang P, McKay BS, Allen JB, et al. Effect of mutant IκB on cytokine-induced activation of NF-κB in cultured human RPE cells[J]. Invest Ophthalmol Vis Sci, 2003, 44(3): 1339-1347. DOI:10.1167/iovs.02-0878.
15. 陈焓, 董晓光, 陈楠, 等. 大鼠视网膜微血管内皮细胞的分离和培养[J]. 眼科新进展, 2005, 25(5): 396-399.DOI:10.3969/j.issn.1003-5141.2005.05.003.Chen H, Dong XG, Chen N, et al. Isolation and culture of rat retinal capillary endothelial cells in vitro [J]. Rec Adv Ophthalmol, 2005, 25(5): 396-399.DOI:10.3969/j.issn.1003-5141.2005.05.003.
16. Wei C, Li L, Gupta S. NF-κB-mediated miR-30b regulation in cardiomyocytes cell death by targeting Bcl-2[J]. Mol Cell Biochem, 2014, 387(1-2): 135-141. DOI: 10.1007/s11010-013-1878-1.
17. Bhat OM, Kumar PU, Giridharan NV, et al. Interleukin-18-induced atherosclerosis involves CD36 and NF-κB crosstalk in Apo E^-/- mice[J]. J Cardiol, 2015, 66(1): 28-35. DOI: 10.1016/j.jjcc.2014.10.012.
18. Plummer SM, Holloway KA, Manson MM, et al. Inhibition of cyclo-oxygenase 2 expression in colon cells by the chemopreventive agent curcumin involves inhibition of NF-κB activation via the NIK/IKK signallingcomplex[J]. Oncogene, 1999, 18(44): 6013-6020. DOI: 10.1038/sj.onc.1202980.
19. DiDonato JA, Mercurio F, Karin M. NF-κB and the link between inflammation and cancer[J]. Immunol Rev, 2012, 246(1): 379-400.DOI: 10.1111/j.1600-065X.2012.01099.x.

Chinese Journal of Ocular Fundus Diseases

The effects of Curcumin on the cellular apoptosis induced by high glucose in rat retinal vascular endothelial cells

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