α-lipoic acid protects against hypoxia/reoxygenation-induced injury in human umbilical vein endothelial cells through suppression of apoptosis and autophagy

Zhang,Jingjing; Deng,Houliang; Liu,Li; Liu,Xiaoxia; Zuo,Xialin; Xu,Qian; Wu,Zhuomin; Peng,Xiaobin; Ji,Aimin

doi:10.3892/mmr.2015.3351

α-lipoic acid protects against hypoxia/reoxygenation-induced injury in human umbilical vein endothelial cells through suppression of apoptosis and autophagy

Authors:
- Jingjing Zhang
- Houliang Deng
- Li Liu
- Xiaoxia Liu
- Xialin Zuo
- Qian Xu
- Zhuomin Wu
- Xiaobin Peng
- Aimin Ji
View Affiliations

Affiliations: Center for Drug Research and Development, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong 510282, P.R. China , Institute of Neurosciences, The Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong 510260, P.R. China, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China
Published online on: February 13, 2015 https://doi.org/10.3892/mmr.2015.3351
Pages: 180-186
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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

α‑lipoic acid (ALA) is known as a powerful antioxidant, which has been reported to have protective effects against various cardiovascular diseases. The present study aimed to determine whether ALA pre‑ or post‑treatment induced protective effects against hypoxia/reoxygenation‑induced injury via inhibition of apoptosis and autophagy in human umbilical vein endothelial cells (HUVECs). In order to simulate the conditions of hypoxia/reoxygenation, HUVECs were subjected to 4 h of oxygen‑glucose deprivation (OGD) followed by 12 h of reoxygenation. For the pre‑treatment, ALA was added to the buffer 12 h prior to OGD, whereas for the post‑treatment, ALA was added at the initiation of reoxygenation. The results demonstrated that ALA pre‑ or post‑treatment significantly reduced lactate dehydrogenase (LDH) release induced through hypoxia/reoxygenation in HUVECs in a dose‑dependent manner; of note, 1 mM ALA pre‑ or post‑treatment exhibited the most potent protective effects. In addition, ALA significantly reduced hypoxia/reoxygenation‑induced loss of mitochondrial membrane potential, apoptosis and the expression of cleaved caspase‑3 in HUVECs. In the presence of the specific autophagy inhibitor 3‑methyladenine, hypoxia/reoxygenation‑induced apoptosis was significantly reduced. Furthermore, the formation of autophagosomes, cytosolic microtubule‑associated protein 1A/1B‑light chain 3 ratio and beclin1 levels significantly increased following hypoxia/reoxygenation injury; however, all of these effects were ameliorated following pre‑ or post‑treatment with ALA. The results of the present study suggested that ALA may provide beneficial protection against hypoxia/reoxygenation‑induced injury via attenuation of apoptosis and autophagy in HUVECs.

View Figures

View References

1	Peyrani P and Ramirez J: What is the association of cardiovascular events with clinical failure in patients with community-acquired pneumonia? Infect Dis Clin North Am. 27:205–210. 2013. View Article : Google Scholar : PubMed/NCBI
2	Navab M, Reddy ST, Van Lenten BJ and Fogelman AM: HDL and cardiovascular disease: atherogenic and atheroprotective mechanisms. Nat Rev Cardiol. 8:222–232. 2011. View Article : Google Scholar : PubMed/NCBI
3	Simbula G, Columbano A, Ledda-Columbano GM, Sanna L, Deidda M, Diana A and Pibiri M: Increased ROS generation and p53 activation in alpha-lipoic acid-induced apoptosis of hepatoma cells. Apoptosis. 12:113–123. 2007. View Article : Google Scholar
4	Kiffin R, Bandyopadhyay U and Cuervo AM: Oxidative stress and autophagy. Antioxid Redox Signal. 8:152–162. 2006. View Article : Google Scholar : PubMed/NCBI
5	Lockshin RA and Zakeri Z: Apoptosis, autophagy and more. Int J Biochem Cell Biol. 36:2405–2419. 2004. View Article : Google Scholar : PubMed/NCBI
6	Kroemer G, Galluzzi L, Vandenabeele P, Abrams J, Alnemri ES, Baehrecke EH, Blagosklonny MV, El-Deiry WS, Golstein P, Green DR, Hengartner M, Knight RA, Kumar S, Lipton SA, Malorni W, Nuňez G, Peter ME, Tschopp J, Yuan J, Piacentini M, Zhivotovsky B and Melino G: Nomenclature Committee on Cell Death 2009: Classification of cell death: recommendations of the nomenclature committee on cell death 2009. Cell Death Differ. 16:3–11. 2009. View Article : Google Scholar :
7	Tang H, Da L, Mao Y, Li Y, Li D, Xu Z, Li F, Wang Y, Tiollais P, Li T and Zhao M: Hepatitis B virus X protein sensitizes cells to starvation-induced autophagy via up-regulation of beclin 1 expression. Hepatology. 49:60–71. 2009. View Article : Google Scholar
8	Balduini W, Carloni S and Buonocore G: Autophagy in hypoxia-ischemia induced brain injury: evidence and speculations. Autophagy. 5:221–233. 2009. View Article : Google Scholar
9	Mizushima N, Levine B, Cuervo AM and Klionsky DJ: Autophagy fights disease through cellular self-digestion. Nature. 451:1069–1075. 2008. View Article : Google Scholar : PubMed/NCBI
10	Gozuacik D and Kimchi A: Autophagy as a cell death and tumor suppressor mechanism. Oncogene. 23:2891–2906. 2004. View Article : Google Scholar : PubMed/NCBI
11	Liberski PP, Gajdusek DC and Brown P: How do neurons degenerate in prion diseases or transmissible spongiform encephalopathies (TSEs): neuronal autophagy revisited. Acta Neurobiol Exp (Wars). 62:141–147. 2002.
12	Lin J, Bierhaus A, Bugert P, Dietrich N, Feng Y, Vom Hagen F, Nawroth P, Brownlee M and Hammes HP: Effect of R-(+)-alpha-lipoic acid on experimental diabetic retinopathy. Diabetologia. 49:1089–1096. 2006. View Article : Google Scholar : PubMed/NCBI
13	Li DW, Li GR, Lu Y, Liu ZQ, Chang M, Yao M, Cheng W and Hu LS: alpha-lipoic acid protects dopaminergic neurons against MPP+-induced apoptosis by attenuating reactive oxygen species formation. Int J Mol Med. 32:108–114. 2013.PubMed/NCBI
14	Cheng TJ, Wang YJ, Kao WW, Chen RJ and Ho YS: Protection against arsenic trioxide-induced autophagic cell death in U118 human glioma cells by use of lipoic acid. Food Chem Toxicol. 45:1027–1038. 2007. View Article : Google Scholar : PubMed/NCBI
15	Karim MR, Fujimura S and Kadowaki M: Vitamin E as a novel enhancer of macroautophagy in rat hepatocytes and H4-II-E cells. Biochem Biophys Res Commun. 394:981–987. 2010. View Article : Google Scholar : PubMed/NCBI
16	Qiu C, Jozsef L, Yu B and Yu J: Saponin monomer 13 of dwarf lilyturf tuber (DT-13) protects serum withdrawal-induced apoptosis through PI3K/Akt in HUVEC. Biochem Biophys Res Commun. 443:74–79. 2014. View Article : Google Scholar
17	Chen YY, Wang X, Li XJ, Xue B, Qu L, Zhang TT, Mu YM and Lu JM: The protective effect of alpha lipoic acid on Schwann cells exposed to constant or intermittent high glucose. Biochem Pharmacol. 84:961–973. 2012. View Article : Google Scholar : PubMed/NCBI
18	Roquer J, Segura T, Serena J and Castillo J: Endothelial dysfunction, vascular disease and stroke: the ARTICO study. Cerebrovasc Dis. 27(Suppl 1): 25–37. 2009. View Article : Google Scholar : PubMed/NCBI
19	Laude K, Thuillez C and Richard V: Coronary endothelial dysfunction after ischemia and reperfusion: a new therapeutic target? Braz J Med Biol Res. 34:1–7. 2001. View Article : Google Scholar : PubMed/NCBI
20	Rahman ST, Merchant N, Haque T, Wahi J, Bhaheetharan S, Ferdinand KC and Khan BV: The impact of lipoic acid on endothelial function and proteinuria in quinapril-treated diabetic patients with stage i hypertension results from the quality study. J Cardiovasc Pharmacol Ther. 17:139–145. 2012. View Article : Google Scholar
21	Bae EH, Lee KS, Lee J, Ma SK, Kim NH, Choi KC, Frøkiaer J, Nielsen S, Kim SY, Kim SZ, Kim SH and Kim SW: Effects of alpha-lipoic acid on ischemia-reperfusion-induced renal dysfunction in rats. Am J Physiol Renal Physiol. 294:F272–F280. 2008. View Article : Google Scholar
22	Chidlow G, Schmidt KG, Wood JP, Melena J and Osborne NN: alpha-lipoic acid protects the retina against ischemia-reperfusion. Neuropharmacology. 43:1015–1025. 2002. View Article : Google Scholar : PubMed/NCBI
23	Schönheit K, Gille L and Nohl H: Effect of alpha-lipoic acid and dihydrolipoic acid on ischemia/reperfusion injury of the heart and heart mitochondria. Biochim Biophys Acta. 1271:335–342. 1995. View Article : Google Scholar : PubMed/NCBI
24	Narasimhan P, Liu J, Song YS, Massengale JL and Chan PH: VEGF Stimulates the ERK 1/2 signaling pathway and apoptosis in cerebral endothelial cells after ischemic conditions. Stroke. 40:1467–1473. 2009. View Article : Google Scholar : PubMed/NCBI
25	Xie R, Li X, Ling Y, Shen C, Wu X, Xu W and Gao X: Alpha-lipoic acid pre- and post-treatments provide protection against in vitro ischemia-reperfusion injury in cerebral endothelial cells via Akt/mTOR signaling. Brain Res. 1482:81–90. 2012. View Article : Google Scholar : PubMed/NCBI
26	Galluzzi L, Morselli E, Kepp O and Kroemer G: Targeting postmitochondrial effectors of apoptosis for neuroprotection. Biochim Biophys Acta. 1787:402–413. 2009. View Article : Google Scholar
27	Chang LK, Putcha GV, Deshmukh M and Johnson EM Jr: Mitochondrial involvement in the point of no return in neuronal apoptosis, Role of caspase-3. Biochimie. 84:223–231. 2002. View Article : Google Scholar : PubMed/NCBI
28	Cao X, Chen A, Yang P, Song X, Liu Y, Li Z, Wang X, Wang L and Li Y: Alpha-lipoic acid protects cardiomyocytes against hypoxia/reoxygenation injury by inhibiting autophagy. Biochem Biophys Res Commun. 441:935–940. 2013. View Article : Google Scholar : PubMed/NCBI
29	Kadowaki M and Karim MR: Cytosolic LC3 ratio as a quantitative index of macroautophagy. Methods Enzymol. 452:199–213. 2009.PubMed/NCBI
30	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
31	Maiuri MC, Le Toumelin G, Criollo A, Rain JC, Gautier F, Juin P, Tasdemir E, Pierron G, Troulinaki K, et al: Functional and physical interaction between Bcl-X(L) and a BH3-like domain in Beclin-1. EMBO J. 26:2443–2605. 2007. View Article : Google Scholar
32	Shimizu S, Konishi A, Nishida Y, Mizuta T, Nishina H, Yamamoto A and Tsujimoto Y: Involvement of JNK in the regulation of autophagic cell death. Oncogene. 29:2070–2082. 2010. View Article : Google Scholar : PubMed/NCBI