Nobiletin protects PC12 cells from ERS‑induced apoptosis in OGD/R injury via activation of the PI3K/AKT pathway

Li,Zi‑Ru; Yang,Lei; Zhen,Jin; Zhao,Yan; Lu,Zu‑Neng

doi:10.3892/etm.2018.6330

Nobiletin protects PC12 cells from ERS‑induced apoptosis in OGD/R injury via activation of the PI3K/AKT pathway

Authors:
- Zi‑Ru Li
- Lei Yang
- Jin Zhen
- Yan Zhao
- Zu‑Neng Lu
View Affiliations

Affiliations: Department of Internal Neurology, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China
Published online on: June 20, 2018 https://doi.org/10.3892/etm.2018.6330
Pages: 1470-1476

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

Nobiletin (NOB) possesses multiple pharmacological effects, but its anti‑apoptotic property has acquired a great deal of attention. Endoplasmic reticulum (ER) stress (ERS)‑induced apoptosis acts as the pivotal aetiology in neuronal oxygen‑glucose deprivation and reoxygenation (OGD/R) injury. The aim of this study focused on whether NOB exerts neuro‑protective effects on OGD/R injury by repressing ERS‑induced apoptosis. The PC12 neuronal cell line was subjected to 4 h OGD and 24 h reoxygenation following NOB treatment. A PI3K/AKT inhibitor (LY294002) was added during the mechanistic experiments. Cell viability, lactate dehydrogenase (LDH) release and apoptosis were determined. Western blotting was used to measure protein expression levels. The results showed that OGD/R caused neuronal damageas exhibited by the increase in LDH release and the reduction of cellular viability. Moreover, ERS‑induced apoptosis was markedly stimulated by OGD/R in PC12 cells, as evidenced by the elevation in the apoptotic rate and protein levels of C/EBP homologous protein/glucose‑regulated protein‑78. However, NOB administration significantly reversed neuronal damage and the ERS‑induced apoptosis in response to OGD/R injury. Mechanistic detections showed that the neuron‑favorable and ERS‑repressing contributions of NOB were, in part, a result of the activation of the PI3K/AKT pathway, which was validated by a specific PI3K/AKT inhibitor (LY294002). Therefore, NOB protects PC12 cells from ERS‑induced apoptosis in OGD/R injury mainly through enhancement of the PI3K/AKT pathway, which may provide a novel therapeutic avenue for the prevention of cerebral ischemia/reperfusion injury.

View Figures

View References

1	Gong L, Tang Y, An R, Lin M, Chen L and Du J: RTN1-C mediates cerebral ischemia/reperfusion injury via ER stress and mitochondria-associated apoptosis pathways. Cell Death Dis. 8:e30802017. View Article : Google Scholar : PubMed/NCBI
2	Cai XH, Li XC, Jin SW, Liang DS, Wen ZW, Cao HC, Mei HF, Wu Y, Lin ZD and Wang LX: Endoplasmic reticulum stress plays critical role in brain damage after chronic intermittent hypoxia in growing rats. Exp Neurol. 257:148–156. 2014. View Article : Google Scholar : PubMed/NCBI
3	Xin Q, Ji B, Cheng B, Wang C, Liu H, Chen X, Chen J and Bai B: Endoplasmic reticulum stress in cerebral ischemia. Neurochem Int. 68:18–27. 2014. View Article : Google Scholar : PubMed/NCBI
4	Qiu B, Hu S, Liu L, Chen M, Wang L, Zeng X and Zhu S: CART attenuates endoplasmic reticulum stress response induced by cerebral ischemia and reperfusion through upregulating BDNF synthesis and secretion. Biochem Biophys Res Commun. 436:655–659. 2013. View Article : Google Scholar : PubMed/NCBI
5	Zhang H, Park JH, Maharjan S, Park JA, Choi KS, Park H, Jeong Y, Ahn JH, Kim IH, Lee JC, et al: Sac-1004, a vascular leakage blocker, reduces cerebral ischemia-reperfusion injury by suppressing blood-brain barrier disruption and inflammation. J Neuroinflammation. 14:1222017. View Article : Google Scholar : PubMed/NCBI
6	Kwon SK, Ahn M, Song HJ, Kang SK, Jung SB, Harsha N, Jee S, Moon JY, Suh KS, Lee SD, et al: Nafamostat mesilate attenuates transient focal ischemia/reperfusion-induced brain injury via the inhibition of endoplasmic reticulum stress. Brain Res. 1627:12–20. 2015. View Article : Google Scholar : PubMed/NCBI
7	Min HM, Wang Y, Ren DY, Cheng X, Li J, Jiang XQ, Min LQ and Bao CF: Protective effect of 2-deoxy-D-glucose on the brain tissue in rat cerebral ischemia-reperfusion models by inhibiting Caspase-apoptotic pathway. Histol Histopathol. 32:57–67. 2017.PubMed/NCBI
8	Nakka VP, Gusain A and Raghubir R: Endoplasmic reticulum stress plays critical role in brain damage after cerebral ischemia/reperfusion in rats. Neurotox Res. 17:189–202. 2010. View Article : Google Scholar : PubMed/NCBI
9	Sp N, Kang DY, Joung YH, Park JH, Kim WS, Lee HK, Song KD, Park YM and Yang YM: Nobiletin inhibits angiogenesis by regulating Src/FAK/STAT3-mediated signaling through PXN in ER⁺ breast cancer cells. Int J Mol Sci. 18:E9352017. View Article : Google Scholar : PubMed/NCBI
10	Wu Y, Zhang W, Li M, Cao D, Yang X and Gong J: Nobiletin ameliorates ischemia-reperfusion injury by suppressing the function of Kupffer cells after liver transplantation in rats. Biomed Pharmacother. 89:732–741. 2017. View Article : Google Scholar : PubMed/NCBI
11	Zhang N, Yang Z, Xiang SZ, Jin YG, Wei WY, Bian ZY, Deng W and Tang QZ: Nobiletin attenuates cardiac dysfunction, oxidative stress, and inflammatory in streptozotocin: Induced diabetic cardiomyopathy. Mol Cell Biochem. 417:87–96. 2016. View Article : Google Scholar : PubMed/NCBI
12	Yasuda N, Ishii T, Oyama D, Fukuta T, Agato Y, Sato A, Shimizu K, Asai T, Asakawa T, Kan T, et al: Neuroprotective effect of nobiletin on cerebral ischemia-reperfusion injury in transient middle cerebral artery-occluded rats. Brain Res. 1559:46–54. 2014. View Article : Google Scholar : PubMed/NCBI
13	Zhang N, Wei WY, Yang Z, Che Y, Jin YG, Liao HH, Wang SS, Deng W and Tang QZ: Nobiletin, a polymethoxy flavonoid, protects against cardiac hypertrophy induced by pressure-overload via inhibition of NAPDH oxidases and endoplasmic reticulum stress. Cell Physiol Biochem. 42:1313–1325. 2017. View Article : Google Scholar : PubMed/NCBI
14	Shi MD, Liao YC, Shih YW and Tsai LY: Nobiletin attenuates metastasis via both ERK and PI3K/Akt pathways in HGF-treated liver cancer HepG2 cells. Phytomedicine. 20:743–752. 2013. View Article : Google Scholar : PubMed/NCBI
15	Lee YC, Cheng TH, Lee JS, Chen JH, Liao YC, Fong Y, Wu CH and Shih YW: Nobiletin, a citrus flavonoid, suppresses invasion and migration involving FAK/PI3K/Akt and small GTPase signals in human gastric adenocarcinoma AGS cells. Mol Cell Biochem. 347:103–115. 2011. View Article : Google Scholar : PubMed/NCBI
16	Zhang JF, Zhang L, Shi LL, Zhao ZH, Xu H, Liang F, Li HB, Zhao Y, Xu X, Yang K and Tian YF: Parthenolide attenuates cerebral ischemia/reperfusion injury via Akt/GSK-3β pathway in PC12 cells. Biomed Pharmacother. 89:1159–1165. 2017. View Article : Google Scholar : PubMed/NCBI
17	Mo ZT, Li WN, Zhai YR and Gao SY: The effects of icariin on the expression of HIF-1α, HSP-60 and HSP-70 in PC12 cells suffered from oxygen-glucose deprivation-induced injury. Pharm Biol. 55:848–852. 2017. View Article : Google Scholar : PubMed/NCBI
18	Liu X, Zhu X, Chen M, Ge Q, Shen Y and Pan S: Resveratrol protects PC12 cells against OGD/R-induced apoptosis via the mitochondrial-mediated signaling pathway. Acta Biochim Biophys Sin (Shanghai). 48:342–353. 2016. View Article : Google Scholar : PubMed/NCBI
19	Wang ZG, Cheng Y, Yu XC, Ye LB, Xia QH, Johnson NR, Wei X, Chen DQ, Cao G, Fu XB, et al: bFGF protects against blood-brain barrier damage through junction protein regulation via PI3K-Akt-Rac1 pathway following traumatic brain injury. Mol Neurobiol. 53:7298–7311. 2016. View Article : Google Scholar : PubMed/NCBI
20	Chen L, Ren Z, Wei X, Wang S, Wang Y, Cheng Y, Gao H and Liu H: Losartan protects against cerebral ischemia/reperfusion-induced apoptosis through β-arrestin1-mediated phosphorylation of Akt. Eur J Pharmacol. 815:98–108. 2017. View Article : Google Scholar : PubMed/NCBI
21	Zhang Q, An R, Tian X, Yang M, Li M, Lou J, Xu L and Dong Z: β-caryophyllene pretreatment alleviates focal cerebral ischemia-reperfusion injury by activating PI3K/Akt signaling pathway. Neurochem Res. 42:1459–1469. 2017. View Article : Google Scholar : PubMed/NCBI
22	Wang Z, Wang Y, Ye J, Lu X, Cheng Y, Xiang L, Chen L, Feng W, Shi H, Yu X, et al: bFGF attenuates endoplasmic reticulum stress and mitochondrial injury on myocardial ischaemia/reperfusion via activation of PI3K/Akt/ERK1/2 pathway. J Cell Mol Med. 19:595–607. 2015. View Article : Google Scholar : PubMed/NCBI
23	Yung HW, Korolchuk S, Tolkovsky AM, Charnock-Jones DS and Burton GJ: Endoplasmic reticulum stress exacerbates ischemia-reperfusion-induced apoptosis through attenuation of Akt protein synthesis in human choriocarcinoma cells. FASEB J. 21:872–884. 2007. View Article : Google Scholar : PubMed/NCBI
24	Yuan Y, Guo Q, Ye Z, Pingping X, Wang N and Song Z: Ischemic postconditioning protects brain from ischemia/reperfusion injury by attenuating endoplasmic reticulum stress-induced apoptosis through PI3K-Akt pathway. Brain Res. 1367:85–93. 2011. View Article : Google Scholar : PubMed/NCBI