Protocatechuic acid attenuates β‑secretase activity and okadaic acid‑induced autophagy via the Akt/GSK‑3β/MEF2D pathway in PC12 cells

Huang,Liping; Zhong,Xiaoqin; Qin,Shaochen; Deng,Minzhen

doi:10.3892/mmr.2019.10905

Protocatechuic acid attenuates β‑secretase activity and okadaic acid‑induced autophagy via the Akt/GSK‑3β/MEF2D pathway in PC12 cells

Authors:
- Liping Huang
- Xiaoqin Zhong
- Shaochen Qin
- Minzhen Deng
View Affiliations

Affiliations: School of Chemistry and Chemical Engineering, Lingnan Normal University, Zhanjiang, Guangdong 524048, P.R. China, Department of Neurology, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou, Guangdong 510120, P.R. China, Department of Neurology, The Affiliated Hospital of Shanxi University of Chinese Medicine, Taiyuan, Shanxi 030024, P.R. China
Published online on: December 30, 2019 https://doi.org/10.3892/mmr.2019.10905
Pages: 1328-1335

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

Okadaic acid (OA) can be used to induce an Alzheimer's disease (AD) model characterized by tau hyperphosphorylation, the formation of neurofibrillary tangles formation and β‑amyloid (Aβ) deposition. Previous studies have shown that the upregulation of Beclin‑1‑dependent autophagy may contribute to the elimination of aggregated Aβ. However, the effects of protocatechuic acid (PA) on the levels of Aβ42, phosphorylated (p)‑tau and β‑secretase in OA‑induced cell injury are unclear, and little is known concerning the role of the PA signaling pathway in the regulation of autophagy. The present study aimed to determine whether PA protects cells from OA‑induced cytotoxicity via the regulation of Beclin‑1‑dependent autophagy and its regulatory signaling pathway. PC12 cells were treated with OA with or without PA for 24 h. Enzymatic assays were performed to measure p‑tau, Aβ42 and β‑secretase activity. Western blotting was performed to detect p‑Akt, p‑glycogen synthase kinase‑3β (p‑GSK‑3β), Akt, GSK‑3β, myocyte enhancer factor 2D (MEF2D) and Beclin‑1 protein expression levels. Immunofluorescence and immunocytochemistry were used to measure Beclin‑1 expression levels. The results from this study showed that PA could increase cell viability and significantly decrease the levels of Aβ42, p‑tau, β‑secretase and Beclin‑1. PA can also promote the expression of p‑Akt and MEF2D while suppressing the expression of p‑GSK‑3β. These results indicated that PA protects PC12 cells from OA‑induced cytotoxicity, and attenuates autophagy via regulation of the Akt/GSK‑3β/MEF2D pathway, therefore potentially contributing to the neuroprotective effects of PA against OA toxicity. These findings suggested that PA may have potential as a drug candidate in preventative AD therapy.

View Figures

View References

1	Reas ET, Hagler DJ Jr, White NS, Kuperman JM, Bartsch H, Cross K, Loi RQ, Balachandra AR, Meloy MJ, Wierenga CE, et al: Sensitivity of restriction spectrum imaging to memory and neuropathology in Alzheimer's disease. Alzheimers Res Ther. 9:552017. View Article : Google Scholar : PubMed/NCBI
2	Lue LF, Guerra A and Walker DG: Amyloid beta and tau as Alzheimer's disease blood biomarkers: Promise from new technologies. Neurol Ther. 6 (Suppl 1):S25–S36. 2017. View Article : Google Scholar
3	Mamada N, Tanokashira D, Ishii K, Tamaoka A and Araki W: Mitochondria are devoid of amyloid β-protein (Aβ)-producing secretases: Evidence for unlikely occurrence within mitochondria of Aβ generation from amyloid precursor protein. Biochem Biophys Res Commun. 486:321–328. 2017. View Article : Google Scholar : PubMed/NCBI
4	Ban JY, Cho SO, Jeon SY, Bae K, Song KS and Seong YH: 3,4-dihydroxybenzoic acid from Smilacis chinae rhizome protects amyloid beta protein (25–35)-induced neurotoxicity in cultured rat cortical neurons. Neurosci Lett. 420:184–188. 2007. View Article : Google Scholar : PubMed/NCBI
5	Xue XY, Zhang Z, Zhou XW, Zhou X and Luo HM: The inhibition effect of protocatechuic acid on the mRNA expression of APP on M146L cell. Zhong Yao Cai. 35:1813–1816. 2012.(In Chinese). PubMed/NCBI
6	Song Y, Cui T, Xie N, Zhang X, Qian Z and Liu J: Protocatechuic acid improves cognitive deficits and attenuates amyloid deposits, inflammatory response in aged AβPP/PS1 double transgenic mice. Int Immunopharmacol. 20:276–281. 2014. View Article : Google Scholar : PubMed/NCBI
7	Todde V, Veenhuis M and van der Klei IJ: Autophagy: Principles and significance in health and disease. Biochim Biophys Acta. 1792:3–13. 2009. View Article : Google Scholar : PubMed/NCBI
8	Qian X, Li X, Cai Q, Zhang C, Yu Q, Jiang Y, Lee JH, Hawke D, Wang Y, Xia Y, et al: Phosphoglycerate kinase 1 phosphorylates beclin1 to induce autophagy. Mol Cell. 65:917–931.e6. 2017. View Article : Google Scholar : PubMed/NCBI
9	Bustos V, Pulina MV, Bispo A, Lam A, Flajolet M, Gorelick FS and Greengard P: Phosphorylated Presenilin 1 decreases β-amyloid by facilitating autophagosome-lysosome fusion. Proc Natl Acad Sci USA. 114:7148–7153. 2017. View Article : Google Scholar : PubMed/NCBI
10	Gu X, Guo W, Zhao Y, Liu G, Wu J and Chang C: Deoxynivalenol-induced cytotoxicity and apoptosis in IPEC-J2 cells through the activation of autophagy by inhibiting PI3K-AKT-mTOR signaling pathway. ACS Omega. 4:18478–18486. 2019. View Article : Google Scholar : PubMed/NCBI
11	Yang Q, She H, Gearing M, Colla E, Lee M, Shacka JJ and Mao Z: Regulation of neuronal survival factor MEF2D by chaperone-mediated autophagy. Science. 323:124–127. 2009. View Article : Google Scholar : PubMed/NCBI
12	Xu H, Li J, Wang Z, Feng M, Shen Y, Cao S, Li T, Peng Y, Fan L, Chen J, et al: Methylene blue attenuates neuroinflammation after subarachnoid hemorrhage in rats through the Akt/GSK-3β/MEF2D signaling pathway. Brain Behav Immun. 65:125–139. 2017. View Article : Google Scholar : PubMed/NCBI
13	Song Z, Feng C, Lu Y, Gao Y, Lin Y and Dong C: Overexpression and biological function of MEF2D in human pancreatic cancer. Am J Transl Res. 9:4836–4847. 2017.PubMed/NCBI
14	Chen ZW, Liu A, Liu Q, Chen J, Li WM, Chao XJ, Yang Q, Liu PQ, Mao ZX and Pi RB: MEF2D mediates the neuroprotective effect of methylene blue against glutamate-induced oxidative damage in HT22 hippocampal cells. Mol Neurobiol. 54:2209–2222. 2017. View Article : Google Scholar : PubMed/NCBI
15	Brayton CF: Dimethyl sulfoxide (DMSO): A review. Cornell Vet. 76:61–90. 1986.PubMed/NCBI
16	Usman MS, Hussein MZ, Kura AU, Fakurazi S, Masarudin MJ and Ahmad Saad FF: Graphene oxide as a nanocarrier for a theranostics delivery system of protocatechuic acid and gadolinium/gold nanoparticles. Molecules. 23(pii): E5002018. View Article : Google Scholar : PubMed/NCBI
17	Huang L, Deng M, He Y, Lu S, Ma R and Fang Y: β-asarone and levodopa co-administration increase striatal dopamine level in 6-hydroxydopamine induced rats by modulating P-glycoprotein and tight junction proteins at the blood-brain barrier and promoting levodopa into the brain. Clin Exp Pharmacol Physiol. 43:634–643. 2016. View Article : Google Scholar : PubMed/NCBI
18	Koehler D, Shah ZA and Williams FE: The GSK3β inhibitor, TDZD-8, rescues cognition in a zebrafish model of okadaic acid-induced Alzheimer's disease. Neurochem Int. 122:31–37. 2019. View Article : Google Scholar : PubMed/NCBI
19	Theendakara V, Bredesen DE and Rao RV: Downregulation of protein phosphatase 2A by apolipoprotein E: Implications for Alzheimer's disease. Mol Cell Neurosci. 83:83–91. 2017. View Article : Google Scholar : PubMed/NCBI
20	Wang Y, Song X, Liu D, Lou YX, Luo P, Zhu T, Wang Q and Chen N: IMM-H004 reduced okadaic acid-induced neurotoxicity by inhibiting Tau pathology in vitro and in vivo. Neurotoxicology. 75:221–232. 2019. View Article : Google Scholar : PubMed/NCBI
21	Krzysztoforska K, Mirowska-Guzel D and Widy-Tyszkiewicz E: Pharmacological effects of protocatechuic acid and its therapeutic potential in neurodegenerative diseases: Review on the basis of in vitro and in vivo studies in rodents and humans. Nutr Neurosci. 22:72–82. 2019. View Article : Google Scholar : PubMed/NCBI
22	Winter AN, Brenner MC, Punessen N, Snodgrass M, Byars C, Arora Y and Linseman DA: Comparison of the neuroprotective and anti-inflammatory effects of the anthocyanin metabolites, protocatechuic acid and 4-hydroxybenzoic acid. Oxid Med Cell Longev. 2017:62970802017. View Article : Google Scholar : PubMed/NCBI
23	Adedara IA, Okpara ES, Busari EO, Omole O, Owumi SE and Farombi EO: Dietary protocatechuic acid abrogates male reproductive dysfunction in streptozotocin-induced diabetic rats via suppression of oxidative damage, inflammation and caspase-3 activity. Eur J Pharmacol. 849:30–42. 2019. View Article : Google Scholar : PubMed/NCBI
24	Tang XL, Liu JX, Dong W, Li P, Li L, Lin CR, Zheng YQ, Cong WH and Hou JC: Cardioprotective effect of protocatechuic acid on myocardial ischemia/reperfusion injury. J Pharmacol Sci. 125:176–183. 2014. View Article : Google Scholar : PubMed/NCBI
25	Yin X, Zhang X, Lv C, Li C, Yu Y, Wang X and Han F: Protocatechuic acid ameliorates neurocognitive functions impairment induced by chronic intermittent hypoxia. Sci Rep. 5:145072015. View Article : Google Scholar : PubMed/NCBI
26	Zeng Q, Siu W, Li L, Jin Y, Liang S, Cao M, Ma M and Wu Z: Autophagy in Alzheimer's disease and promising modulatory effects of herbal medicine. Exp Gerontol. 119:100–110. 2019. View Article : Google Scholar : PubMed/NCBI
27	Huang L, Lin M, Zhong X, Yang H and Deng M: Galangin decreases p-tau, Aβ42 and β-secretase levels, and suppresses autophagy in okadaic acid-induced PC12 cells via an Akt/GSK3β/mTOR signaling-dependent mechanism. Mol Med Rep. 19:1767–1774. 2019.PubMed/NCBI
28	Deng M, Huang L, Ning B, Wang N, Zhang Q, Zhu C and Fang Y: β-asarone improves learning and memory and reduces Acetyl Cholinesterase and Beta-amyloid 42 levels in APP/PS1 transgenic mice by regulating Beclin-1-dependent autophagy. Brain Res. 1652:188–194. 2016. View Article : Google Scholar : PubMed/NCBI
29	Zhang S, Gui XH, Huang LP, Deng MZ, Fang RM, Ke XH, He YP, Li L and Fang YQ: Neuroprotective effects of β-asarone against 6-hydroxy dopamine-induced parkinsonism via JNK/Bcl-2/Beclin-1 pathway. Mol Neurobiol. 53:83–94. 2016. View Article : Google Scholar : PubMed/NCBI
30	Huang L, Deng M, He Y, Lu S, Liu S and Fang Y: β-asarone increases MEF2D and TH levels and reduces α-synuclein level in 6-OHDA-induced rats via regulating the HSP70/MAPK/MEF2D/Beclin-1 pathway: Chaperone-mediated autophagy activation, macroautophagy inhibition and HSP70 up-expression. Behav Brain Res. 313:370–379. 2016. View Article : Google Scholar : PubMed/NCBI
31	Reddy PH, Yin X, Manczak M, Kumar S, Pradeepkiran JA, Vijayan M and Reddy AP: Mutant APP and amyloid beta-induced defective autophagy, mitophagy, mitochondrial structural and functional changes and synaptic damage in hippocampal neurons from Alzheimer's disease. Hum Mol Genet. 27:2502–2516. 2018. View Article : Google Scholar : PubMed/NCBI
32	Song J, Ma SJ, Luo JH, Zhang H, Wang RX, Liu H, Li L, Zhang ZG and Zhou RX: Melatonin induces the apoptosis and inhibits the proliferation of human gastric cancer cells via blockade of the AKT/MDM2 pathway. Oncol Rep. 39:1975–1983. 2018.PubMed/NCBI
33	Zhang LY, Liu ZH, Zhu Q, Wen S, Yang CX, Fu ZJ and Sun T: Resolvin D2 relieving radicular pain is associated with regulation of inflammatory mediators, Akt/GSK-3β signal pathway and GPR18. Neurochem Res. 43:2384–2392. 2018. View Article : Google Scholar : PubMed/NCBI
34	Yao Y, Wang Y, Kong L, Chen Y and Yang J: Osthole decreases tau protein phosphorylation via PI3K/AKT/GSK-3β signaling pathway in Alzheimer's disease. Life Sci. 217:16–24. 2019. View Article : Google Scholar : PubMed/NCBI
35	Hu S, Cui W, Zhang Z, Mak S, Xu D, Li G, Hu Y, Wang Y, Lee M, Tsim KW and Han Y: Indirubin-3-oxime effectively prevents 6OHDA-induced neurotoxicity in PC12 cells via activating MEF2D through the inhibition of GSK3β. J Mol Neurosci. 57:561–570. 2015. View Article : Google Scholar : PubMed/NCBI