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Published on: April 12, 2015
MicroRNA-128-3p Affects Neuronal Apoptosis and Neurobehavior in Cerebral Palsy Rats by Targeting E3 Ubiquitin-Linking
Xiaoqi Nie1, Rui Cheng1, Pengfei Hao1,2
1Department of Neurosurgery, Shanxi Provincial People's Hospital, No. 29, Shuangta East Street, Yingze District, Taiyuan, 030012, Shanxi, China.
Abstract:
This study was dedicated to investigating the effects of microRNA-128-3p (miR-128-3p) on neuronal apoptosis and neurobehavior in cerebral palsy (CP) rats via the Smurf2/YY1 axis.In vivo modeling of hypoxic-ischemic (HI) CP was established in neonatal rats. Neurobehavioral tests (geotaxis reflex, cliff avoidance reaction, and grip test) were measured after HI induction. The HI-induced neurological injury was evaluated by HE staining, Nissl staining, TUNEL staining, immunohistochemical staining, and RT-qPCR. The expression of miR-128-3p, Smurf2, and YY1 was determined by RT-qPCR and western blot techniques. Moreover, primary cortical neurons were used to establish the oxygen and glucose deprivation (OGD) model in vitro, cell viability was detected by CCK-8 assay, neuronal apoptosis was assessed by flow cytometry and western blot, and the underlying mechanism between miR-128-3p, Smurf2 and YY1 was verified by bioinformatics analysis, dual luciferase reporter assay, RIP, Co-IP, ubiquitination assay, western blot, and RT-qPCR.In vivo, miR-128-3p and YY1 expression was elevated, and Smurf2 expression was decreased in brain tissues of hypoxic-ischemic CP rats. Downregulation of miR-128-3p or overexpression of Smurf2 improved neurobehavioral performance, reduced neuronal apoptosis, and elevated Nestin and NGF expression in hypoxic-ischemic CP rats, and downregulation of Smurf2 reversed the effects of downregulation of miR-128-3p on neurobehavioral performance, neuronal apoptosis, and Nestin and NGF expression in hypoxic-ischemic CP rats, while overexpression of YY1 reversed the effects of Smurf2 on neurobehavioral performance, neuronal apoptosis, and Nestin and NGF expression in hypoxic-ischemic CP rats. In vitro, downregulation of miR-128-3p effectively promoted the neuronal survival, reduced the apoptosis rate, and decreased caspase3 protein expression after OGD, and overexpression of YY1 reversed the ameliorative effect of downregulation of miR-128-3p on OGD-induced neuronal injury. miR-128-3p targeted to suppress Smurf2 to lower YY1 ubiquitination degradation and decrease its expression.Inhibition of miR-128-3p improves neuronal apoptosis and neurobehavioral changes in hypoxic-ischemic CP rats by promoting Smurf2 to promote YY1 ubiquitination degradation and reduce YY1 expression.
Insights
Inhibition of microRNA-128-3p (miR-128-3p) improves neurobehavior and reduces neuronal apoptosis in cerebral palsy rats. This occurs by enhancing Smurf2, which promotes YY1 degradation, thereby mitigating neurological injury.
Area of Science:
- Neuroscience
- Molecular Biology
- Genetics
Background:
- Cerebral palsy (CP) is a debilitating neurological disorder often resulting from hypoxic-ischemic (HI) injury.
- Understanding the molecular mechanisms underlying neuronal apoptosis and neurobehavioral deficits in CP is crucial for developing effective treatments.
Purpose of the Study:
- To investigate the role of microRNA-128-3p (miR-128-3p) in neuronal apoptosis and neurobehavioral outcomes in a rat model of HI-induced CP.
- To elucidate the molecular pathway involving Smurf2 and YY1 that mediates the effects of miR-128-3p in CP.
Main Methods:
- Establishment of an in vivo HI model in neonatal rats and an in vitro oxygen-glucose deprivation (OGD) model using primary cortical neurons.
- Assessment of neurobehavioral performance, neurological injury (histopathology, apoptosis assays), and expression levels of miR-128-3p, Smurf2, and YY1 using molecular and biochemical techniques.
- Mechanistic investigations including bioinformatics analysis, dual luciferase reporter assays, RIP, Co-IP, and ubiquitination assays to confirm the interactions between miR-128-3p, Smurf2, and YY1.
Main Results:
- In HI-CP rats, miR-128-3p and YY1 expression were elevated, while Smurf2 expression was decreased.
- Downregulation of miR-128-3p or overexpression of Smurf2 ameliorated neurobehavioral deficits, reduced neuronal apoptosis, and increased Nestin and NGF expression in HI-CP rats.
- miR-128-3p directly targets Smurf2, suppressing its expression by reducing YY1 ubiquitination and degradation. Overexpression of YY1 reversed the protective effects of miR-128-3p downregulation.
Conclusions:
- miR-128-3p plays a detrimental role in HI-induced CP by suppressing Smurf2 and consequently increasing YY1 expression, leading to enhanced neuronal apoptosis and impaired neurobehavior.
- Inhibition of miR-128-3p represents a potential therapeutic strategy for CP by restoring the Smurf2/YY1 balance, reducing neuronal injury, and improving neurological function.
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