Naringenin upregulates GTPCH1/eNOS to ameliorate high glucose‑induced retinal endothelial cell injury

Xue,Bing; Wang,Yu

doi:10.3892/etm.2022.11355

Naringenin upregulates GTPCH1/eNOS to ameliorate high glucose‑induced retinal endothelial cell injury

Authors:
- Bing Xue
- Yu Wang
View Affiliations

Affiliations: Health Management Center of Dalian Second People's Hospital, Dalian, Liaoning 116011, P.R. China, Medical Department of Dalian Second People's Hospital, Dalian, Liaoning 116011, P.R. China
Published online on: May 5, 2022 https://doi.org/10.3892/etm.2022.11355
Article Number: 428
Copyright: © Xue et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Diabetic retinopathy (DR) is a microvascular complication of diabetes, while retinal endothelial cell (REC) dysfunction is considered the primary pathological process of DR. Naringenin, a natural flavonoid compound, exhibits therapeutic potential against multiple types of endothelial cell injury. To the best of our knowledge, however, its effect on REC injury has not been previously investigated. Therefore, the aim of the present study was to investigate the effect of naringenin on high glucose (HG)‑induced REC injury and assess the underlying mechanism. To establish a retinal injury model, human (H)RECs were treated with 30 mM glucose. Cell Counting Kit‑8 assay and TUNEL staining were used to assess the effects of naringenin on cell proliferation and apoptosis, respectively. Reactive oxygen species (ROS) levels and concentration of tetrahydrobiopterin (BH4), the essential cofactor of endothelial nitric oxide synthase (eNOS), were measured using a ROS detection kit and ELISA, respectively. The transfection efficiency of HRECs with guanosine triphosphate cyclohydrolase‑1 (GTPCH1) interfering plasmid was examined by reverse transcription‑quantitative PCR. The protein expression levels of Ki67, proliferative cell nuclear antigen (PCNA), eNOS and GTPCH1 were determined by western blot analysis. Compared with the HG‑induced group alone, co‑treatment with naringenin inhibited HG‑induced HREC apoptosis in a dose‑dependent manner, increased expression levels of the proliferation‑associated proteins Ki67 and PCNA and effectively decreased intracellular ROS levels. Furthermore, naringenin upregulated GTPCH1/eNOS signaling and promoted release of BH4. However, GTPCH1 knockdown partially reversed the ameliorative effect of naringenin on HG‑induced HREC injury. In summary, the present study suggested that naringenin effectively inhibited HG‑induced HREC apoptosis and intracellular oxidative stress, which may be associated with naringenin‑mediated GTPCH1/eNOS upregulation.

View Figures

View References

1	Wojciechowska J, Krajewski W, Bolanowski M, Kręcicki T and Zatoński T: Diabetes and cancer: A review of current knowledge. Exp Clin Endocrinol Diabetes. 124:263–275. 2016.PubMed/NCBI View Article : Google Scholar
2	Li Y and Ren K: The mechanism of contrast-induced acute kidney injury and its association with diabetes mellitus. Contrast Media Mol Imaging. 2020(3295176)2020.PubMed/NCBI View Article : Google Scholar
3	Basu P and Basu A: In vitro and in vivo effects of flavonoids on peripheral neuropathic pain. Molecules. 25(1171)2020.PubMed/NCBI View Article : Google Scholar
4	Piano I, Di Paolo M, Corsi F, Piragine E, Bisti S, Gargini C and Di Marco S: Retinal neurodegeneration: Correlation between nutraceutical treatment and animal model. Nutrients. 13(770)2021.PubMed/NCBI View Article : Google Scholar
5	Lechner J, O'Leary OE and Stitt AW: The pathology associated with diabetic retinopathy. Vision Res. 139:7–14. 2017.PubMed/NCBI View Article : Google Scholar
6	Hammes HP: Diabetic retinopathy: Hyperglycaemia, oxidative stress and beyond. Diabetologia. 61:29–38. 2018.PubMed/NCBI View Article : Google Scholar
7	Yang Y, Liu Y, Li Y, Chen Z, Xiong Y, Zhou T, Tao W, Xu F, Yang H, Ylä-Herttuala S, et al: MicroRNA-15b targets VEGF and inhibits angiogenesis in proliferative diabetic retinopathy. J Clin Endocrinol Metab. 105:3404–3415. 2020.PubMed/NCBI View Article : Google Scholar
8	Fu D, Yu JY, Yang S, Wu M, Hammad SM, Connell AR, Du M, Chen J and Lyons TJ: Survival or death: A dual role for autophagy in stress-induced pericyte loss in diabetic retinopathy. Diabetologia. 59:2251–2261. 2016.PubMed/NCBI View Article : Google Scholar
9	Hua YQ, Zeng Y, Xu J and Xu XL: Naringenin alleviates nonalcoholic steatohepatitis in middle-aged Apoe^-/-mice: Role of SIRT1. Phytomedicine. 81(153412)2021.PubMed/NCBI View Article : Google Scholar
10	Fuster MG, Carissimi G, Montalbán MG and Víllora G: Improving anticancer therapy with naringenin-loaded silk fibroin nanoparticles. Nanomaterials (Basel). 10(718)2020.PubMed/NCBI View Article : Google Scholar
11	Hernández-Aquino E and Muriel P: Beneficial effects of naringenin in liver diseases: Molecular mechanisms. World J Gastroenterol. 24:1679–1707. 2018.PubMed/NCBI View Article : Google Scholar
12	Naraki K, Rezaee R and Karimi G: A review on the protective effects of naringenin against natural and chemical toxic agents. Phytother Res. 35:4075–4091. 2021.PubMed/NCBI View Article : Google Scholar
13	Patel K, Singh GK and Patel DK: A review on pharmacological and analytical aspects of naringenin. Chin J Integr Med. 24:551–560. 2018.PubMed/NCBI View Article : Google Scholar
14	Tutunchi H, Naeini F, Ostadrahimi A and Hosseinzadeh-Attar MJ: Naringenin, a flavanone with antiviral and anti-inflammatory effects: A promising treatment strategy against COVID-19. Phytother Res. 34:3137–3147. 2020.PubMed/NCBI View Article : Google Scholar
15	Al-Dosari DI, Ahmed MM, Al-Rejaie SS, Alhomida AS and Ola MS: Flavonoid naringenin attenuates oxidative stress, apoptosis and improves neurotrophic effects in the diabetic rat retina. Nutrients. 9(1161)2017.PubMed/NCBI View Article : Google Scholar
16	Heidary Moghaddam R, Samimi Z, Moradi SZ, Little PJ, Xu S and Farzaei MH: Naringenin and naringin in cardiovascular disease prevention: A preclinical review. Eur J Pharmacol. 887(173535)2020.PubMed/NCBI View Article : Google Scholar
17	Burke AC, Sutherland BG, Telford DE, Morrow MR, Sawyez CG, Edwards JY and Huff MW: Naringenin enhances the regression of atherosclerosis induced by a chow diet in Ldlr^-/- mice. Atherosclerosis. 286:60–70. 2019.PubMed/NCBI View Article : Google Scholar
18	Qurtam AA, Mechchate H, Es-Safi I, Al-Zharani M, Nasr FA, Noman OM, Aleissa M, Imtara H, Aleissa AM, Bouhrim M and Alqahtani AS: Citrus flavanone narirutin, in vitro and in silico mechanistic antidiabetic potential. Pharmaceutics. 13(1818)2021.PubMed/NCBI View Article : Google Scholar
19	Zeng B, Chen K, Du P, Wang SS, Ren B, Ren YL, Yan HS, Liang Y and Wu FH: Phenolic compounds from clinopodium chinense (Benth.) O. Kuntze and their inhibitory effects on α-glucosidase and vascular endothelial cells injury. Chem Biodivers. 13:596–601. 2016.PubMed/NCBI View Article : Google Scholar
20	Oguido APMT, Hohmann MSN, Pinho-Ribeiro FA, Crespigio J, Domiciano TP, Verri WA Jr and Casella AMB: Naringenin eye drops inhibit corneal neovascularization by anti-inflammatory and antioxidant mechanisms. Invest Ophthalmol Vis Sci. 58:5764–5776. 2017.PubMed/NCBI View Article : Google Scholar
21	Nasser A, Møller AT, Hellmund V, Thorborg SS, Jespersgaard C, Bjerrum OJ, Dupont E, Nachman G, Lykkesfeldt J, Jensen TS and Møller LB: Heterozygous mutations in GTP-cyclohydrolase-1 reduce BH4 biosynthesis but not pain sensitivity. Pain. 159:1012–1024. 2018.PubMed/NCBI View Article : Google Scholar
22	Liu P, Liu J, Wu Y, Xi W, Wei Y, Yuan Z and Zhuo X: Zinc supplementation protects against diabetic endothelial dysfunction via GTP cyclohydrolase 1 restoration. Biochem Biophys Res Commun. 521:1049–1054. 2020.PubMed/NCBI View Article : Google Scholar
23	Liang YH, Chen GW, Li XS, Jia S and Meng CY: Guanosine-5'-triphosphate cyclohydrolase 1 regulated long noncoding RNAs are potential targets for microglial activation in neuropathic pain. Neural Regen Res. 16:596–600. 2021.PubMed/NCBI View Article : Google Scholar
24	Li J, Liu S, Cao G, Sun Y, Chen W, Dong F, Xu J, Zhang C and Zhang W: Nicotine induces endothelial dysfunction and promotes atherosclerosis via GTPCH1. J Cell Mol Med. 22:5406–5417. 2018.PubMed/NCBI View Article : Google Scholar
25	Sandrim VC, Yugar-Toledo JC, Desta Z, Flockhart DA, Moreno H Jr and Tanus-Santos JE: Endothelial nitric oxide synthase haplotypes are related to blood pressure elevation, but not to resistance to antihypertensive drug therapy. J Hypertens. 24:2393–2397. 2006.PubMed/NCBI View Article : Google Scholar
26	Satoh M, Fujimoto S, Haruna Y, Arakawa S, Horike H, Komai N, Sasaki T, Tsujioka K, Makino H and Kashihara N: NAD(P)H oxidase and uncoupled nitric oxide synthase are major sources of glomerular superoxide in rats with experimental diabetic nephropathy. Am J Physiol Renal Physiol. 288:F1144–F1152. 2005.PubMed/NCBI View Article : Google Scholar
27	Wang S, Xu J, Song P, Wu Y, Zhang J, Chul Choi H and Zou MH: Acute inhibition of guanosine triphosphate cyclohydrolase 1 uncouples endothelial nitric oxide synthase and elevates blood pressure. Hypertension. 52:484–490. 2008.PubMed/NCBI View Article : Google Scholar
28	Alp NJ, McAteer MA, Khoo J, Choudhury RP and Channon KM: Increased endothelial tetrahydrobiopterin synthesis by targeted transgenic GTP-cyclohydrolase I overexpression reduces endothelial dysfunction and atherosclerosis in ApoE-knockout mice. Arterioscler Thromb Vasc Biol. 24:445–450. 2004.PubMed/NCBI View Article : Google Scholar
29	Pannirselvam M, Simon V, Verma S, Anderson T and Triggle CR: Chronic oral supplementation with sepiapterin prevents endothelial dysfunction and oxidative stress in small mesenteric arteries from diabetic (db/db) mice. Br J Pharmacol. 140:701–706. 2003.PubMed/NCBI View Article : Google Scholar
30	Meininger CJ, Marinos RS, Hatakeyama K, Martinez-Zaguilan R, Rojas JD, Kelly KA and Wu G: Impaired nitric oxide production in coronary endothelial cells of the spontaneously diabetic BB rat is due to tetrahydrobiopterin deficiency. Biochem J. 349:353–356. 2000.PubMed/NCBI View Article : Google Scholar
31	Liu L, Zhang H, Shi Y and Pan L: Prostaglandin E1 improves cerebral microcirculation through activation of endothelial NOS and GRPCH1. J Mol Neurosci. 70:2041–2048. 2020.PubMed/NCBI View Article : Google Scholar
32	Le Y, Wei R, Yang K, Lang S, Gu L, Liu J, Hong T and Yang J: Liraglutide ameliorates palmitate-induced oxidative injury in islet microvascular endothelial cells through GLP-1 receptor/PKA and GTPCH1/eNOS signaling pathways. Peptides. 124(170212)2020.PubMed/NCBI View Article : Google Scholar
33	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar
34	Rizwan H, Pal S, Sabnam S and Pal A: High glucose augments ROS generation regulates mitochondrial dysfunction and apoptosis via stress signalling cascades in keratinocytes. Life Sci. 241(117148)2020.PubMed/NCBI View Article : Google Scholar
35	Wang S, Xu J, Song P, Viollet B and Zou MH: In vivo activation of AMP-activated protein kinase attenuates diabetes-enhanced degradation of GTP cyclohydrolase I. Diabetes. 58:1893–1901. 2009.PubMed/NCBI View Article : Google Scholar
36	Li RF, Feng YQ, Chen JH, Ge LT, Xiao SY and Zuo XL: Naringenin suppresses K562 human leukemia cell proliferation and ameliorates Adriamycin-induced oxidative damage in polymorphonuclear leukocytes. Exp Ther Med. 9:697–706. 2015.PubMed/NCBI View Article : Google Scholar
37	Li H, Chen P, Chen L and Wang X: The natural flavonoid naringenin inhibits the cell growth of wilms tumor in children by suppressing TLR4/NF-κB signaling. Anticancer Agents Med Chem. 21:1120–1126. 2021.PubMed/NCBI View Article : Google Scholar
38	Ahsan H: Diabetic retinopathy-biomolecules and multiple pathophysiology. Diabetes Metab Syndr. 9:51–54. 2015.PubMed/NCBI View Article : Google Scholar
39	Simó R, Stitt AW and Gardner TW: Neurodegeneration in diabetic retinopathy: Does it really matter? Diabetologia. 61:1902–1912. 2018.PubMed/NCBI View Article : Google Scholar
40	Jin H, Zhu Y, Li Y, Ding X, Ma W, Han X and Wang B: BDNF-mediated mitophagy alleviates high-glucose-induced brain microvascular endothelial cell injury. Apoptosis. 24:511–528. 2019.PubMed/NCBI View Article : Google Scholar
41	Cao Y, Yuan G, Zhang Y and Lu R: High glucose-induced circHIPK3 downregulation mediates endothelial cell injury. Biochem Biophys Res Commun. 507:362–368. 2018.PubMed/NCBI View Article : Google Scholar
42	Ding X, Yao W, Zhu J, Mu K, Zhang J and Zhang JA: Resveratrol attenuates high glucose-induced vascular endothelial cell injury by activating the E2F3 pathway. Biomed Res Int. 2020(6173618)2020.PubMed/NCBI View Article : Google Scholar
43	Zhang Y, Lv X, Hu Z, Ye X, Zheng X, Ding Y, Xie P and Liu Q: Protection of Mcc950 against high-glucose-induced human retinal endothelial cell dysfunction. Cell Death Dis. 8(e2941)2017.PubMed/NCBI View Article : Google Scholar
44	Long L, Li Y, Yu S, Li X, Hu Y, Long T, Wang L, Li W, Ye X, Ke Z and Xiao H: Scutellarin prevents angiogenesis in diabetic retinopathy by downregulating VEGF/ERK/FAK/Src pathway signaling. J Diabetes Res. 2019(4875421)2019.PubMed/NCBI View Article : Google Scholar
45	Wojnar W, Zych M and Kaczmarczyk-Sedlak I: Antioxidative effect of flavonoid naringenin in the lenses of type 1 diabetic rats. Biomed Pharmacother. 108:974–984. 2018.PubMed/NCBI View Article : Google Scholar
46	Chen W and Lin B, Xie S, Yang W, Lin J, Li Z, Zhan Y, Gui S and Lin B: Naringenin protects RPE cells from NaIO₃-induced oxidative damage in vivo and in vitro through up-regulation of SIRT1. Phytomedicine. 80(153375)2021.PubMed/NCBI View Article : Google Scholar
47	Meza CA, La Favor JD, Kim DH and Hickner RC: Endothelial dysfunction: Is there a hyperglycemia-induced imbalance of NOX and NOS? Int J Mol Sci. 20(3775)2019.PubMed/NCBI View Article : Google Scholar
48	Kim DH, Meza CA, Clarke H, Kim JS and Hickner RC: Vitamin D and endothelial function. Nutrients. 12(575)2020.PubMed/NCBI View Article : Google Scholar
49	Gielis JF, Lin JY, Wingler K, Van Schil PE, Schmidt HH and Moens AL: Pathogenetic role of eNOS uncoupling in cardiopulmonary disorders. Free Radic Biol Med. 50:765–776. 2011.PubMed/NCBI View Article : Google Scholar
50	Wu Y, Ding Y, Ramprasath T and Zou MH: Oxidative stress, GTPCH1, and endothelial nitric oxide synthase uncoupling in hypertension. Antioxid Redox Signal. 34:750–764. 2021.PubMed/NCBI View Article : Google Scholar
51	An H, Wei R, Ke J, Yang J, Liu Y, Wang X, Wang G and Hong T: Metformin attenuates fluctuating glucose-induced endothelial dysfunction through enhancing GTPCH1-mediated eNOS recoupling and inhibiting NADPH oxidase. J Diabetes Complications. 30:1017–1024. 2016.PubMed/NCBI View Article : Google Scholar
52	Bai Y, Peng W, Yang C, Zou W, Liu M, Wu H, Fan L, Li P, Zeng X and Su W: Pharmacokinetics and metabolism of naringin and active metabolite naringenin in rats, dogs, humans, and the differences between species. Front Pharmacol. 11(364)2020.PubMed/NCBI View Article : Google Scholar