Neural Regeneration Research ›› 2019, Vol. 14 ›› Issue (9): 1603-1609.doi: 10.4103/1673-5374.255979

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miR-15b-5p targeting amyloid precursor protein is involved in the anti-amyloid eflect of curcumin in swAPP695-HEK293 cells

Hong-Ying Liu, Xian Fu, You-Fu Li, Xian-Liang Li, Zhen-Yu Ma, Ying Zhang, Qing-Chun Gao   

  1. Department of Neurology, the Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong Province, China
  • Online:2019-09-15 Published:2019-09-15
  • Contact: Qing-Chun Gao, MD, qcgao@263.net.
  • Supported by:

    his study was supported by the Science and Technology Planning Project of Guangdong Province of China, No. 2016A020226022 (to HYL); the Medical and Health Technology Project of Guangzhou of China, No. 20161A011068 (to HYL); the Guangzhou Science Technology and Innovation Commission of China, No.201704020043 (to QCG).

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Abstract:

Curcumin exerts a neuroprotective effect on Alzheimer’s disease; however, it is not known whether microRNAs are involved in this pro¬tective effect. This study was conducted using swAPP695-HEK293 cells as an Alzheimer’s disease cell model. swAPP695-HEK293 cells were treated with 0, 0.5, 1, 2, 5, and 10 μM curcumin for 24 hours. The changes in miR-15b-5p, miR-19a-3p, miR-195-5p, miR-101-3p, miR-216b-5p, miR-16-5p and miR-185-5p expression were assessed by real-time quantitative polymerase chain reaction. The mRNA and protein levels of amyloid precursor protein, amyloid-β40 and amyloid-β42 were evaluated by quantitative real-time polymerase chain re¬action, western blot assays and enzyme-linked immunosorbent assays. swAPP695-HEK293 cells were transfected with miR-15b-5p mimic, or treated with 1 μM curcumin 24 hours before miR-15b-5p inhibitor transfection. The effects of curcumin on amyloid precursor protein, amyloid-β40 and amyloid-β42 levels were evaluated by western blot assays and enzyme-linked immunosorbent assay. Luciferase assays were used to analyze the interaction between miR-15b-5p and the 3′-untranslated region of amyloid precursor protein. The results show that amyloid precursor protein and amyloid-β expression were enhanced in swAPP695-HEK293 cells compared with HEK293 parental cells. Curcumin suppressed the expression of amyloid precursor protein and amyloid-β and up-regulated the expression of miR-15b-5p in swAPP695-HEK293 cells. In addition, we found a negative association of miR-15b-5p expression with amyloid precursor protein and amyloid-β levels in the curcumin-treated cells. Luciferase assays revealed that miR-15b-5p impaired the luciferase activity of the plasmid harboring the 3′-untranslated region of amyloid precursor protein. These findings indicate that curcumin down-regulates the expression of amyloid precursor protein and amyloid-β in swAPP695-HEK293 cells, which was partially mediated by miR-15b-5p via targeting of the 3′-untranslated region of amyloid precursor protein.

Key words: nerve regeneration, Alzheimer’s disease, natural plant drug, curcuminoids, miRNAs, amyloid precursor protein, amyloid-β, 3′-untranslated region, Luciferase assays, neurons, neural regeneration