Isoquercitrin protects HUVECs against high glucose‑induced apoptosis through regulating p53 proteasomal degradation

Liu,Libo; Huang,Sihui; Xu,Man; Gong,Yan; Li,Dan; Wan,Chunxia; Wu,Haiming; Tang,Qizhu

doi:10.3892/ijmm.2021.4955

Isoquercitrin protects HUVECs against high glucose‑induced apoptosis through regulating p53 proteasomal degradation

Authors:
- Libo Liu
- Sihui Huang
- Man Xu
- Yan Gong
- Dan Li
- Chunxia Wan
- Haiming Wu
- Qizhu Tang
View Affiliations

Affiliations: Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China, Pharmacy Department, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, P.R. China
Published online on: May 7, 2021 https://doi.org/10.3892/ijmm.2021.4955
Article Number: 122
Copyright: © Liu 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

High glucose (HG)‑induced endothelial apoptosis serves an important role in the vascular dysfunction associated with diabetes mellitus (DM). It has been reported that isoquercitrin (IQC), a flavonoid glucoside, possesses an anti‑DM effect, but the mechanism requires further investigation. The present study investigated the effect of IQC against HG‑induced apoptosis in human umbilical vein endothelial cells (HUVECs) and explored its molecular mechanism. HUVECs were treated with 5 or 30 mM glucose for 48 h. Endothelial cell viability was monitored using the Cell Counting Kit‑8 assay. Mitochondrial membrane potential was detected by JC‑1 staining. Apoptosis was observed by TUNEL staining and flow cytometry. Western blotting was used for the analysis of apoptosis‑associated proteins Bax, Bcl‑2, cleaved (C)‑caspase3, total‑caspase3, p53 and phosphorylated p53. Reverse transcription‑quantitative PCR was used to analyze the mRNA expression levels of Bax, Bcl‑2 and p53. Immunofluorescence staining was utilized to detect the expression levels and distribution of p53 and ubiquitin specific peptidase 10 (USP10) in HUVECs. The results revealed that IQC significantly attenuated HG‑induced endothelial apoptosis, as shown by decreased apoptotic cells observed by TUNEL, JC‑1 staining and flow cytometry. Moreover, under HG stress, IQC treatment markedly inhibited the increased expression levels of the pro‑apoptotic proteins p53, Bax and C‑caspase3, and increased the expression levels of the anti‑apoptotic protein Bcl‑2 in HUVECs. However, the anti‑apoptotic effect of IQC against HG was partially blunted by increasing p53 protein levels in vitro. IQC influenced the mRNA expression levels of Bax and Bcl‑2 in response to HG, but it did not affect the transcription of p53. Notably, IQC inhibited the HG‑induced phosphorylation of p53 at Ser15 and the nuclear transport of USP10, destabilizing p53 and increasing the proteasomal degradation of the p53 protein. The current findings revealed that IQC exerted a protective effect against the HG‑induced apoptosis of endothelial cells by regulating the proteasomal degradation of the p53 protein, suggesting that IQC may be used as a novel therapeutic compound to ameliorate DM‑induced vascular complications.

View Figures

View References

1	Schmidt AM: Highlighting diabetes mellitus: The epidemic continues. Arterioscler Thromb Vasc Biol. 38:e1–e8. 2018. View Article : Google Scholar
2	Eelen G, de Zeeuw P, Simons M and Carmeliet P: Endothelial cell metabolism in normal and diseased vasculature. Circ Res. 116:1231–1244. 2015. View Article : Google Scholar : PubMed/NCBI
3	Yu S, Liu X, Men L, Yao J, Xing Q and Du J: Selenoprotein S protects against high glucose-induced vascular endothelial apoptosis through the PKCβII/JNK/Bcl-2 pathway. J Cell Biochem. Nov 28–2018.Online ahead of print.
4	Watson EC, Grant ZL and Coultas L: Endothelial cell apoptosis in angiogenesis and vessel regression. Cell Mol Life Sci. 74:4387–4403. 2017. View Article : Google Scholar : PubMed/NCBI
5	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:e29412017. View Article : Google Scholar : PubMed/NCBI
6	Han X, Wang B, Sun Y, Huang J, Wang X, Ma W, Zhu Y, Xu R, Jin H and Liu N: Metformin modulates high glucose-incubated human umbilical vein endothelial cells proliferation and apoptosis through AMPK/CREB/BDNF pathway. Front Pharmacol. 9:12662018. View Article : Google Scholar : PubMed/NCBI
7	Gu J, Huang W, Zhang W, Zhao T, Gao C, Gan W, Rao M, Chen Q, Guo M, Xu Y and Xu YH: Sodium butyrate alleviates high-glucose-induced renal glomerular endothelial cells damage via inhibiting pyroptosis. Int Immunopharmacol. 75:1058322019. View Article : Google Scholar : PubMed/NCBI
8	Zhao X, Su L, He X, Zhao B and Miao J: Long noncoding RNA CA7-4 promotes autophagy and apoptosis via sponging MIR877-3P and MIR5680 in high glucose-induced vascular endothelial cells. Autophagy. 16:70–85. 2020. View Article : Google Scholar :
9	Ko YS, Jin H, Park SW and Kim HJ: Salvianolic acid B protects against oxLDL-induced endothelial dysfunction under high-glucose conditions by downregulating ROCK1-mediated mitophagy and apoptosis. Biochem Pharmacol. 174:1138152020. View Article : Google Scholar
10	Wei H, Cao C, Wei X, Meng M, Wu B, Meng L, Wei X, Gu S and Li H: Circular RNA circVEGFC accelerates high glucose-induced vascular endothelial cells apoptosis through miR-338-3p/HIF-1α/VEGFA axis. Aging (Albany NY). 12:14365–14375. 2020. View Article : Google Scholar
11	Yi J and Gao ZF: MicroRNA-9-5p promotes angiogenesis but inhibits apoptosis and inflammation of high glucose-induced injury in human umbilical vascular endothelial cells by targeting CXCR4. Int J Biol Macromol. 130:1–9. 2019. View Article : Google Scholar : PubMed/NCBI
12	Valentová K, Vrba J, Bancířová M, Ulrichová J and Křen V: Isoquercitrin: Pharmacology, toxicology, and metabolism. Food Chem Toxicol. 68:267–282. 2014. View Article : Google Scholar : PubMed/NCBI
13	Huang SH, Xu M, Wu HM, Wan CX, Wang HB, Wu QQ, Liao HH, Deng W and Tang QZ: Isoquercitrin attenuated cardiac dysfunction via AMPKα-dependent pathways in LPS-treated mice. Mol Nutr Food Res. 62:e18009552018. View Article : Google Scholar
14	Ma C, Jiang Y, Zhang X, Chen X, Liu Z and Tian X: Isoquercetin ameliorates myocardial infarction through anti-inflammation and anti-apoptosis factor and regulating TLR4-NF-κB signal pathway. Mol Med Rep. 17:6675–6680. 2018.PubMed/NCBI
15	Gasparotto Junior A, Dos Reis Piornedo R, Assreuy J and Da Silva-Santos JE: Nitric oxide and Kir6.1 potassium channel mediate isoquercitrin-induced endothelium-dependent and independent vasodilation in the mesenteric arterial bed of rats. Eur J Pharmacol. 788:328–334. 2016. View Article : Google Scholar : PubMed/NCBI
16	Bondonno NP, Bondonno CP, Ward NC, Woodman RJ, Hodgson JM and Croft KD: Enzymatically modified isoquercitrin improves endothelial function in volunteers at risk of cardiovascular disease. Br J Nutr. 123:182–189. 2020. View Article : Google Scholar
17	Jayachandran M, Wu Z, Ganesan K, Khalid S, Chung SM and Xu B: Isoquercetin upregulates antioxidant genes, suppresses inflammatory cytokines and regulates AMPK pathway in streptozotocin-induced diabetic rats. Chem Biol Interact. 303:62–69. 2019. View Article : Google Scholar : PubMed/NCBI
18	Niu C, Chen Z, Kim KT, Sun J, Xue M, Chen G, Li S, Shen Y, Zhu Z, Wang X, et al: Metformin alleviates hyperglycemia-induced endothelial impairment by downregulating autophagy via the hedgehog pathway. Autophagy. 15:843–870. 2019. View Article : Google Scholar : PubMed/NCBI
19	Yu L, Liang Q, Zhang W, Liao M, Wen M, Zhan B, Bao H and Cheng X: HSP22 suppresses diabetes-induced endothelial injury by inhibiting mitochondrial reactive oxygen species formation. Redox Biol. 21:1010952019. View Article : Google Scholar : PubMed/NCBI
20	Liu J, Meng Z, Gan L, Guo R, Gao J, Liu C, Zhu D, Liu D, Zhang L, Zhang Z, et al: C1q/TNF-related protein 5 contributes to diabetic vascular endothelium dysfunction through promoting Nox-1 signaling. Redox Biol. 34:1014762020. View Article : Google Scholar : PubMed/NCBI
21	Christodoulou MS, Colombo F, Passarella D, Ieronimo G, Zuco V, De Cesare M and Zunino F: Synthesis and biological evaluation of imidazolo(2,1-b)benzothiazole derivatives, as potential p53 inhibitors. Bioorg Med Chem. 19:1649–1657. 2011. View Article : Google Scholar : PubMed/NCBI
22	Zhang Y, Zhang Q, Zeng SX, Zhang Y, Mayo LD and Lu H: Inauhzin and Nutlin3 synergistically activate p53 and suppress tumor growth. Cancer Biol Ther. 13:915–924. 2012. View Article : Google Scholar : PubMed/NCBI
23	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. View Article : Google Scholar
24	Kwon SK, Saindane M and Baek KH: p53 stability is regulated by diverse deubiquitinating enzymes. Biochim Biophys Acta Rev Cancer. 1868:404–411. 2017. View Article : Google Scholar : PubMed/NCBI
25	Domingueti CP, Dusse LM, Carvalho MD, de Sousa LP, Gomes KB and Fernandes AP: Diabetes mellitus: The linkage between oxidative stress, inflammation, hypercoagulability and vascular complications. J Diabetes Complications. 30:738–745. 2016. View Article : Google Scholar : PubMed/NCBI
26	Elshaer SL, Lemtalsi T and El-Remessy AB: High glucose-mediated tyrosine nitration of PI3-Kinase: A molecular switch of survival and apoptosis in endothelial cells. Antioxidants (Basel). 7:472018. View Article : Google Scholar
27	Liu C, Yao MD, Li CP, Shan K, Yang H, Wang JJ, Liu B, Li XM, Yao J, Jiang Q and Yan B: Silencing of circular RNA-ZNF609 ameliorates vascular endothelial dysfunction. Theranostics. 7:2863–2877. 2017. View Article : Google Scholar : PubMed/NCBI
28	Kim JH, Lee S and Cho EJ: The protective effects of acer okamotoanum and Isoquercitrin on obesity and amyloidosis in a mouse model. Nutrients. 12:13532020. View Article : Google Scholar :
29	Resham K, Khare P, Bishnoi M and Sharma SS: Neuroprotective effects of isoquercitrin in diabetic neuropathy via Wnt/β-catenin signaling pathway inhibition. Biofactors. 46:411–420. 2020. View Article : Google Scholar : PubMed/NCBI
30	Chen L, Feng P, Peng A, Qiu X, Lai W, Zhang L and Li W: Protective effects of isoquercitrin on streptozotocin-induced neurotoxicity. J Cell Mol Med. 24:10458–10467. 2020. View Article : Google Scholar : PubMed/NCBI
31	Dai Y, Zhang H, Zhang J and Yan M: Isoquercetin attenuates oxidative stress and neuronal apoptosis after ischemia/reperfusion injury via Nrf2-mediated inhibition of the NOX4/ROS/NF-κB pathway. Chem Biol Interact. 284:32–40. 2018. View Article : Google Scholar : PubMed/NCBI
32	Dong Y, Chen H, Gao J, Liu Y, Li J and Wang J: Molecular machinery and interplay of apoptosis and autophagy in coronary heart disease. J Mol Cell Cardiol. 136:27–41. 2019. View Article : Google Scholar : PubMed/NCBI
33	Bykov VJ, Issaeva N, Shilov A, Hultcrantz M, Pugacheva E, Chumakov P, Bergman J, Wiman KG and Selivanova G: Restoration of the tumor suppressor function to mutant p53 by a low-molecular-weight compound. Nat Med. 8:282–288. 2002. View Article : Google Scholar : PubMed/NCBI
34	Gao Y, Yin H, Zhang Y, Dong Y, Yang F, Wu X and Liu H: Dexmedetomidine protects hippocampal neurons against hypoxia/reoxygenation-induced apoptosis through activation HIF-1α/p53 signaling. Life Sci. 232:1166112019. View Article : Google Scholar
35	Schisano B, Tripathi G, McGee K, McTernan PG and Ceriello A: Glucose oscillations, more than constant high glucose, induce p53 activation and a metabolic memory in human endothelial cells. Diabetologia. 54:1219–1226. 2011. View Article : Google Scholar : PubMed/NCBI
36	Li Q, Pang L, Shi H, Yang W, Liu X, Su G and Dong Y: High glucose concentration induces retinal endothelial cell apoptosis by activating p53 signaling pathway. Int J Clin Exp Pathol. 11:2401–2407. 2018.PubMed/NCBI
37	Wu Y, Lee S, Bobadilla S, Duan SZ and Liu X: High glucose-induced p53 phosphorylation contributes to impairment of endothelial antioxidant system. Biochim Biophys Acta Mol Basis Dis. 1863:2355–2362. 2017. View Article : Google Scholar : PubMed/NCBI
38	Chan WH and Wu HJ: Methylglyoxal and high glucose co-treatment induces apoptosis or necrosis in human umbilical vein endothelial cells. J Cell Biochem. 103:1144–1157. 2008. View Article : Google Scholar
39	Si R, Zhang Q, Tsuji-Hosokawa A, Watanabe M, Willson C, Lai N, Wang J, Dai A, Scott BT, Dillmann WH, et al: Overexpression of p53 due to excess protein O-GlcNAcylation is associated with coronary microvascular disease in type 2 diabetes. Cardiovasc Res. 116:1186–1198. 2020. View Article : Google Scholar :
40	Ao H, Liu B, Li H and Lu L: Egr1 mediates retinal vascular dysfunction in diabetes mellitus via promoting p53 transcription. J Cell Mol Med. 23:3345–3356. 2019. View Article : Google Scholar : PubMed/NCBI
41	Da Pozzo E, La Pietra V, Cosimelli B, Da Settimo F, Giacomelli C, Marinelli L, Martini C, Novellino E, Taliani S and Greco G: p53 functional inhibitors behaving like pifithrin-β counteract the Alzheimer peptide non-β-amyloid component effects in human SH-SY5Y cells. ACS Chem Neurosci. 5:390–399. 2014. View Article : Google Scholar : PubMed/NCBI
42	Chen DZ, Wang WW, Chen YL, Yang XF, Zhao M and Yang YY: miR-128 is upregulated in epilepsy and promotes apoptosis through the SIRT1 cascade. Int J Mol Med. 44:694–704. 2019.PubMed/NCBI
43	Kelly RM, Goren EM, Taylor PA, Mueller SN, Stefanski HE, Osborn MJ, Scott HS, Komarova EA, Gudkov AV, Holländer GA and Blazar BR: Short-term inhibition of p53 combined with keratinocyte growth factor improves thymic epithelial cell recovery and enhances T-cell reconstitution after murine bone marrow transplantation. Blood. 115:1088–1097. 2010. View Article : Google Scholar :
44	Jung SH, Kim BJ, Lee EH and Osborne NN: Isoquercitrin is the most effective antioxidant in the plant Thuja orientalis and able to counteract oxidative-induced damage to a transformed cell line (RGC-5 cells). Neurochem Int. 57:713–721. 2010. View Article : Google Scholar : PubMed/NCBI
45	Taha KF, Khalil M, Abubakr MS and Shawky E: Identifying cancer-related molecular targets of Nandina domestica Thunb. by network pharmacology-based analysis in combination with chemical profiling and molecular docking studies. J Ethnopharmacol. 249:1124132020. View Article : Google Scholar
46	Jung JH, Lee H, Zeng SX and Lu H: RBM10, a new regulator of p53. Cells. 9:21072020. View Article : Google Scholar :
47	Saint-Germain E, Mignacca L, Vernier M, Bobbala D, Ilangumaran S and Ferbeyre G: SOCS1 regulates senescence and ferroptosis by modulating the expression of p53 target genes. Aging (Albany NY). 9:2137–2162. 2017. View Article : Google Scholar
48	Liu Y, Tavana O and Gu W: p53 modifications: Exquisite decorations of the powerful guardian. J Mol Cell Biol. 11:564–577. 2019. View Article : Google Scholar : PubMed/NCBI
49	Shieh SY, Ikeda M, Taya Y and Prives C: DNA damage-induced phosphorylation of p53 alleviates inhibition by MDM2. Cell. 91:325–334. 1997. View Article : Google Scholar : PubMed/NCBI
50	Momand J, Zambetti GP, Olson DC, George D and Levine AJ: The mdm-2 oncogene product forms a complex with the p53 protein and inhibits p53-mediated transactivation. Cell. 69:1237–1245. 1992. View Article : Google Scholar : PubMed/NCBI
51	Liu J, Xia H, Kim M, Xu L, Li Y, Zhang L, Cai Y, Norberg HV, Zhang T, Furuya T, et al: Beclin1 controls the levels of p53 by regulating the deubiquitination activity of USP10 and USP13. Cell. 147:223–234. 2011. View Article : Google Scholar : PubMed/NCBI
52	Yuan J, Luo K, Zhang L, Cheville JC and Lou Z: USP10 regulates p53 localization and stability by deubiquitinating p53. Cell. 140:384–396. 2010. View Article : Google Scholar : PubMed/NCBI
53	Deng CC, Zhu DH, Chen YJ, Huang TY, Peng Y, Liu SY, Lu P, Xue YH, Xu YP, Yang B and Rong Z: TRAF4 promotes fibroblast proliferation in keloids by destabilizing p53 via interacting with the deubiquitinase USP10. J Invest Dermatol. 139:1925–1935.e5. 2019. View Article : Google Scholar
54	Honda R, Tanaka H and Yasuda H: Oncoprotein MDM2 is a ubiquitin ligase E3 for tumor suppressor p53. FEBS Lett. 420:25–27. 1997. View Article : Google Scholar
55	Carmona V, Martín-Aragón S, Goldberg J, Schubert D and Bermejo-Bescós P: Several targets involved in Alzheimer's disease amyloidogenesis are affected by morin and isoquercitrin. Nutr Neurosci. 23:575–590. 2020. View Article : Google Scholar