Protein disulfide isomerase A3 might be involved in the regulation of 24-dehydrocholesterol reductase via vitamin D equilibrium in primary cortical neurons

YAVUZ U., ALAYLIOĞLU M., Sengul B., Karras S. N., GEZEN AK D., DURSUN E.

IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL, cilt.57, ss.704-714, 2021 (SCI-Expanded, Scopus) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 57
  • Basım Tarihi: 2021
  • Doi Numarası: 10.1007/s11626-021-00602-5
  • Dergi Adı: IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, BIOSIS, Biotechnology Research Abstracts, CAB Abstracts, EMBASE, MEDLINE, Veterinary Science Database
  • Sayfa Sayıları: ss.704-714
  • Anahtar Kelimeler: Vitamin D, Neurodegeneration, 24-dehydrocholesterol reductase, Protein disulfide isomerase A3, Primary neuronal culture, AMYLOID-BETA 1-42, CENTRAL-NERVOUS-SYSTEM, ALZHEIMERS-DISEASE, CHOLESTEROL-BIOSYNTHESIS, D DEFICIENCY, 1,25-DIHYDROXYVITAMIN D-3, OXIDATIVE STRESS, IN-VITRO, EXPRESSION, RECEPTOR
  • İstanbul Üniversitesi-Cerrahpaşa Adresli: Evet

Özet

Vitamin D is a secosteroid hormone mediating its functions via vitamin D receptor (VDR) and an endoplasmic reticulum chaperone, protein disulfide isomerase A3 (PDIA3). From a physiological perspective, there is also a well-established association of cholesterol and vitamin D synthesis, since both share a common metabolic substrate, 7 dehydrocholesterol (7-DHC). Yet, the potential basic pathways, of the biological interplay of DHCR24 and vitamin D equilibrium, on neuronal level, are yet to be determined. In this study, we aimed to investigate the relation between vitamin D pathways and DHCR24 in primary cortical neuron cultures. The neocortex of Sprague-Dawley rat embryos (E16) was used for the preparation of primary cortical neuron cultures. DHCR24 mRNA and protein expression levels were determined by qRT-PCR, Western blotting, and immunofluorescent labeling in 1,25-dihydroxyvitamin D3-treated or VDR/PDIA3-silenced primary cortical neurons. The mRNA expression of DHCR24 was significantly decreased in the cortical neurons treated with 10(-8)M 1,25-dihydroxyvitamin D-3 (p<0.001). In parallel with the mRNA results, DHCR24 protein expression in cortical neurons treated with 10(-8)M 1,25-dihydroxyvitamin D-3 was also significantly lower than untreated neurons (p<0.05). These data were also confirmed with immunofluorescent labeling and fluorescence intensity measurements of DHCR24 (p<0.001). Finally, DHCR24 mRNA expression level was significantly increased in PDIA3 siRNA-treated neurons (p<0.05). Similar to the mRNA results, the DHCR24 protein expression of PDIA3 siRNA-treated neurons was also statistically higher than the other groups (p<0.05). Results of this mechanistic experimental basic study demonstrate that DHCR24 mRNA expression and protein concentrations attenuated in response to vitamin D treatment. Furthermore, we observed that PDIA3 might be involved in this modulatory effect. Our findings indicate a complex interaction of DHCR24 and vitamin D equilibrium, through the involvement of PDIA3 and vitamin D in the modulation of cholesterol metabolism in neuronal cells, requiring future studies on the field.