Neural Regeneration Research ›› 2022, Vol. 17 ›› Issue (1): 170-177.doi: 10.4103/1673-5374.314314

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Ghrelin alleviates 6-hydroxydopamine-induced neurotoxicity in SH-SY5Y cells

Xin He1, 2, Wei Yuan3, Chun-Qing Yang4, Lu Zhu1, Fei Liu2, Juan Feng2, Yi-Xue Xue1, *   

  1. 1Department of Neurobiology, School of Life Sciences, China Medical University, Shenyang, Liaoning Province, China; 2Department of Neurology, Shengjing Hospital of China Medical University, Shenyang, Liaoning Province, China; 3Department of Orthopedics, First Hospital of China Medical University, Shenyang, Liaoning Province, China; 4Department of Neurosurgery, Shengjing Hospital of China Medical University, Shenyang, Liaoning Province, China
  • Online:2022-01-15 Published:2021-09-22
  • Contact: Yi-Xue Xue, PhD, xyxcmu@hotmail.com or xueyixue888@163.com.
  • Supported by:
    This work was supported by the National Natural Science Foundation of China, No. 81901417 (to XH); the Natural Science Foundation Doctoral Research Initiation Plan of Liaoning Province of China, No. 2019-BS-287 (to XH); the China Postdoctoral Science Foundation, No. 2019M661173 (to XH).

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Abstract: Ghrelin is a neuropeptide that has various physiological functions and has been demonstrated to be neuroprotective in a number of neurological disease models. However, the underlying mechanisms of ghrelin in Parkinson’s disease remain largely unexplored. The current study aimed to study the effects of ghrelin in a 6-hydroxydopamine (6-OHDA)-induced Parkinson’s disease model and evaluate the potential underlying mechanisms. In the present study, we treated an SH-SY5Y cell model with 6-OHDA, and observed that pretreatment with different concentrations of ghrelin (1, 10, and 100 nM) for 30 minutes relieved the neurotoxic effects of 6-OHDA, as revealed by Cell Counting Kit-8 and Annexin V/propidium iodide (PI) apoptosis assays. Reverse transcription quantitative polymerase chain reaction and western blot assay results demonstrated that 6-OHDA treatment upregulated α-synuclein and lincRNA-p21 and downregulated TG-interacting factor 1 (TGIF1), which was predicted as a potential transcription regulator of the gene encoding α-synuclein (SNCA). Ghrelin pretreatment was able to reverse the trends caused by 6-OHDA. The Annexin V/PI apoptosis assay results revealed that inhibiting either α-synuclein or lincRNA-p21 expression with small interfering RNA (siRNA) relieved 6-OHDA-induced cell apoptosis. Furthermore, inhibiting lincRNA-p21 also partially upregulated TGIF1. By retrieving information from a bioinformatics database and performing both double luciferase and RNA immunoprecipitation assays, we found that lincRNA-p21 and TGIF1 were able to form a double-stranded RNA-binding protein Staufen homolog 1 (STAU1) binding site and further activate the STAU1-mediated mRNA decay pathway. In addition, TGIF1 was able to transcriptionally regulate α-synuclein expression by binding to the promoter of SNCA. The Annexin V/PI apoptosis assay results showed that either knockdown of TGIF1 or overexpression of lincRNA-p21 notably abolished the neuroprotective effects of ghrelin against 6-OHDA-induced neurotoxicity. Collectively, these findings suggest that ghrelin exerts neuroprotective effects against 6-OHDA-induced neurotoxicity via the lincRNA-p21/TGIF1/α-synuclein pathway. 

Key words: 6-hydroxydopamine, apoptosis, ghrelin, lincRNA-p21, neuropeptide, neurotoxicity, Parkinson’s disease, STAU1-mediated mRNA decay, TGIF1, α-synuclein