Title:Overexpression of MiR-188-5p Downregulates IL6ST/STAT3/ NLRP3 Pathway to Ameliorate Neuron Injury in Oxygen-glucose Deprivation/Reoxygenation
Volume: 21
Issue: 3
Author(s): Yujie Hu, Ganlan Wang and Guoshuai Yang*
Affiliation:
- Department of Neurology, Central South University Xiangya School of Medicine Affiliated Haikou Hospital, Haikou,
China
Keywords:
MiR-188-5p, inflammation, IL6ST/STAT3, oxygen-glucose deprivation/reoxygenation, cerebral ischemiareperfusion, ameliorate neuron injury.
Abstract:
Background: CI/R, characterized by ischemic injury following abrupt reestablishment
of blood flow, can cause oxidative stress, mitochondrial dysfunction, and apoptosis.
We used oxygen-glucose deprivation/reoxygenation (OGD/R) induced injury in HT22
and primary mouse cortical neurons (MCN) as a model for CI/R.
Objective: This study investigates the role of miR-188-5p in hippocampal neuron cell injury
associated with Cerebral Ischemia-Reperfusion (CI/R).
Methods: HT22 and MCN cells were induced by OGD/R to construct an in vitro model of
CI/R. Cell apoptosis and proliferation were assessed using flow cytometry and the Cell
Counting Kit-8 (CCK8). ELISA was conducted to measure the levels of IL-1β, IL-6, and
TNF-α. Moreover, the interaction between miR-188-5p and IL6ST was investigated using
dual luciferase assay, the expression of miR-188-5p, Bax, cleaved-caspase3, IL-6, Bcl-2,
IL-1β, TNF-α, IL6ST, NFκB, NLRP3 and STAT3 was evaluated using RT-qPCR or Western
blot, and immunofluorescence was used to analyze the co-expression of p-STAT3 and
NLRP3 in neuronal cells.
Results: OGD/R reduced proliferation and miR-188-5p levels and increased IL6ST expression,
inflammation, and apoptosis in HT22 and MCN cells. Moreover, miR-188-5p was
found to bind to IL6ST. Mimics of miR-188-5p reduced apoptosis, lowered the expression
of cleaved-caspase3 and Bax proteins, and elevated Bcl-2 protein expression in cells treated
with OGD/R. Overexpression of miR-188-5p decreased the levels of NLRP3 and p-STAT3
in the OGD/R group. Furthermore, the overexpression of miR-188-5p reduced IL6ST, p-
NFκB/NFκB, p-STAT3/STAT3, and NLRP3 proteins in OGD/R, and these effects could be
reversed by IL6ST overexpression.
Conclusion: Mimics of miR-188-5p were found to inhibit inflammation and the
STAT3/NLRP3 pathway via IL6ST, thereby ameliorating injury in HT22 and MCN cells
treated with OGD/R in the context of CI/R.
