GSK-3β inhibition protects human nucleus pulposus cell against oxidative stress-inducing apoptosis through
Kai Zhu1,2,3, Song Guo4, Guoyi Han2
1Department of Orthopaedics, Shanghai East Hospital, Tongji University School of Medicine, No.150 Jimo Road, Shanghai, 200120, People's Republic of China.
Background:
Oxidative stress in the intervertebral disc leads to nucleus pulposus (NP) degeneration by inducing cell apoptosis. However, the molecular mechanisms underlying this process remain unclear. Increasing evidence indicates that GSK-3β is related to cell apoptosis induced by oxidative stress. In this study, we explored whether GSK-3β inhibition protects human NP cell against apoptosis under oxidative stress.
Methods And Results:
Immunofluorescence staining was used to show the expression of GSK-3β in human NP cells (NPCs). Flow cytometry, mitochondrial staining and western blot (WB) were used to detect apoptosis of treated NPCs, changes of mitochondrial membrane potential and the expression of mitochondrial apoptosis-related proteins using GSK-3β specific inhibitor SB216763. Co-Immunoprecipitation (Co-IP) was used to demonstrate the interaction between GSK-3β and Bcl-2. We delineated the protective effect of GSK-3β specific inhibitor SB216763 on human NPCs apoptosis induced by oxidative stress in vitro. Further, we showed SB216763 exert the protective effect by preservation of the mitochondrial membrane potential and inhibition of caspase 3/7 activity during oxidative injury. The detailed mechanism underlying the antiapoptotic effect of GSK-3β inhibition was also studied by analyzing mitochondrial apoptosis pathway in vitro.
Conclusions:
We concluded that the GSK-3β inhibitor SB216763 protected mitochondrial membrane potential to delay nucleus pulposus cell apoptosis by inhibiting the interaction between GSK-3β and Bcl-2 and subsequently reducing cytochrome c(Cyto-C) release and caspase-3 activation. Together, inhibition of GSK-3β using SB216763 in NPCs may be a favorable therapeutic strategy to slow intervertebral disc degeneration.
Insights
Inhibiting GSK-3β with SB216763 protects nucleus pulposus cells from oxidative stress-induced apoptosis. This strategy preserves mitochondrial function and may slow intervertebral disc degeneration.
Area of Science:
- Biochemistry
- Cell Biology
- Regenerative Medicine
Background:
- Oxidative stress contributes to nucleus pulposus (NP) degeneration by triggering cell apoptosis.
- The precise molecular mechanisms of this process are not fully understood.
- Glycogen synthase kinase-3 beta (GSK-3β) is implicated in oxidative stress-induced apoptosis.
Purpose of the Study:
- To investigate whether inhibiting GSK-3β protects human NP cells against oxidative stress-induced apoptosis.
- To elucidate the underlying molecular mechanisms of GSK-3β inhibition's protective effects.
Main Methods:
- Human NP cells (NPCs) were treated with the GSK-3β specific inhibitor SB216763.
- Apoptosis, mitochondrial membrane potential, and apoptosis-related proteins were assessed using flow cytometry and Western blot.
- Co-immunoprecipitation (Co-IP) was employed to study the interaction between GSK-3β and Bcl-2.
Main Results:
- SB216763 demonstrated a protective effect against oxidative stress-induced apoptosis in human NPCs.
- The inhibitor preserved mitochondrial membrane potential and inhibited caspase 3/7 activity.
- GSK-3β inhibition was shown to affect the mitochondrial apoptosis pathway.
Conclusions:
- GSK-3β inhibitor SB216763 protects NPCs by maintaining mitochondrial membrane potential, inhibiting GSK-3β and Bcl-2 interaction, and reducing cytochrome c release and caspase-3 activation.
- This suggests that inhibiting GSK-3β may be a viable therapeutic approach for intervertebral disc degeneration.
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