miR-125a-5p is involved in microglial activation by regulating HIF-1α through targeting HIF1AN
Journal Title: Journal of Air Force Medical University - Year 2023, Vol 44, Issue 4
Abstract
Objective To observe the effect of miR-125a-5p on microglial activation in BV-2 microglial inflammation model by regulating HIF-1α through targeting HIF1AN. Methods First, BV-2 microglial inflammation model was established with LPS 150 μg / L + IFN-γ 20 μg / L. BV-2 microglia were divided into control group ( Con group) and microglial inflammation model group ( LPS + IFN-γ group) . The expressions of miR-125a-5p and HIF1AN mRNA were detected by real-time quantitative PCR, and the protein expressions of iNOS and HIF-1α were detected by Western blotting. Then, by regulating the expressions of miR-125a-5p and HIF-1α, the expression level of miR-125a-5p and the mRNA expression level of HIF-1α, pro-inflammatory factors (including IL-1β and TNF-α) and M1 polarization marker iNOS were detected by fluorescence quantitative PCR. The protein expression levels of HIF1AN and HIF-1α were detected by Western blotting. Results Compared with Con group, the mRNA expressions of miR-125a-5p, iNOS, IL-1β and TNF-α increased (P < 0. 05), the mRNA expression of HIF1AN decreased ( P < 0. 05), and the protein expressions of HIF-1α and iNOS increased (P < 0. 05) in LPS + IFN-γ group. After up-regulation of miR-125a-5p expression, compared with NC agomir + LPS + IFN-γ group, the mRNA expressions of miR-125a-5p, IL-1β, TNF-α and iNOS increased (P < 0. 05), the protein expression of HIF1AN decreased (P < 0. 05), and the protein expression of HIF-1α increased (P < 0. 05) in miR agomir + LPS + IFN-γ group. After down-regulation of miR-125a-5p expression, compared with NC antagomir + LPS + IFN-γ group, the mRNA expressions of miR-125a-5p, IL-1β, TNF-α and iNOS decreased (P < 0. 05), the protein expression of HIF1AN increased (P < 0. 05), and the protein expression of HIF-1α decreased (P < 0. 05) in miR antagomir + LPS + IFN-γ group. After knockdown of HIF-1α expression, the results showed that compared with NC + LPS + IFN-γ group, the mRNA expressions of HIF-1α, IL-1β, TNF-α and iNOS decreased ( P < 0. 05 ) in HIF-1α siRNA + LPS + IFN-γ group. Conclusion miR-125a-5p is involved in BV-2 microglial activation by regulating HIF-1α through targeting HIF1AN.
Authors and Affiliations
JING Ziwei, FAN Zhao, LI Guangyao, LI Jiangjing, LYU Miaomiao, WEI Shunmin, ZHANG Xiajing, GAO Changjun,SUN Xude
Keywords
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