Related Experiment Video
Updated: Sep 30, 2025
Monitoring Astrocyte Reactivity and Proliferation in Vitro Under Ischemic-Like Conditions
Published on: October 21, 2017
Propofol Rescued Astrocytes from LPS-induced Inflammatory Response via Blocking LncRNA-MEG3/NF-κB Axis
Fan Zhang1, Zhihua Wang2, Bei Sun1
1Department of Anesthesiology, Xiangya Hospital of Central South University, Changsha, 410008, Hunan Province, China.
Objective:
Evidences demonstrate that propofol attenuates neuro-inflammation following brain ischemia. Moreover, LncRNA-MEG3 has been identified as an independent prognostic marker for ischemic stroke patients, and found to correlate to cerebral ischemia in animal models. Therefore, the current study explored the role of propofol in lipopolysaccharide (LPS)-mediated inflammation in cultured astrocytes, along with the molecular mechanism involved in LncRNAMEG3/ NF-κB axis.
Methods:
The primary cultured astrocytes isolated from rats were used to establish an inflammatory model, which were treated with LPS. Propofol was administrated to the primary cultured astrocytes during LPS treatment. The effects of propofol on pro-inflammatory cytokines and the LncRNAMEG3/ NF-κB pathway were detected by ELISA, qRT-PCR and Western Blot assay, respectively. Then, dual-luciferase assay, chromatin immunoprecipitation and RNA immunoprecipitation were used to determine the interaction between LncRNA-MEG3 and NF-κB.
Results:
Our study found propofol to significantly reduce LncRNA-MEG3 expression, which was elevated in LPS-stimulated astrocytes. Moreover, both propofol and LncRNA-MEG3 knockdown remarkably alleviated LPS-induced cytotoxicity by suppressing expressions and release of proinflammatory cytokines. Loss of LncRNA-MEG3 notably suppressed the NF-κB activity and its phosphorylated activation. Additionally, it was also observed that LncRNA-MEG3 could bind nuclear p65/p50, and promote the binding of NF-κB to IL-6 and TNF-α promoters in the nucleus, subsequently stimulating the production of inflammatory cytokines in LPS-treated astrocytes. Furthermore, a specific inhibitor of NF-κB, PDTC, rescued astrocytes from LPS exposure without affecting the LncRNA-MEG3 expression.
Conclusion:
These findings demonstrate that LncRNA-MEG3 acts as a positive regulator of NF-κB, mediating the neuroprotection of propofol in LPS-triggered astrocytes injury.
Insights
Propofol reduces neuroinflammation by downregulating LncRNA-MEG3, which positively regulates the NF-κB pathway. This mechanism protects astrocytes from lipopolysaccharide (LPS)-induced injury.
Area of Science:
- Neuroscience
- Molecular Biology
- Pharmacology
Background:
- Propofol is known to reduce neuroinflammation after brain ischemia.
- Long non-coding RNA-MEG3 (LncRNA-MEG3) is a prognostic marker for ischemic stroke and correlates with cerebral ischemia.
- The molecular mechanisms underlying propofol's neuroprotective effects, particularly involving LncRNA-MEG3 and the NF-κB pathway, require further elucidation.
Purpose of the Study:
- To investigate the role of propofol in lipopolysaccharide (LPS)-induced inflammation in cultured astrocytes.
- To explore the molecular mechanism involving the LncRNA-MEG3/NF-κB axis in propofol's anti-inflammatory effects.
Main Methods:
- Primary rat astrocytes were stimulated with LPS to establish an inflammatory model.
- Propofol was administered, and its effects on pro-inflammatory cytokines and the LncRNA-MEG3/NF-κB pathway were assessed using ELISA, qRT-PCR, and Western Blot.
- Interactions between LncRNA-MEG3 and NF-κB were determined via dual-luciferase, chromatin immunoprecipitation, and RNA immunoprecipitation assays.
Main Results:
- Propofol significantly reduced elevated LncRNA-MEG3 expression in LPS-stimulated astrocytes.
- Both propofol treatment and LncRNA-MEG3 knockdown alleviated LPS-induced cytotoxicity by suppressing pro-inflammatory cytokine release.
- LncRNA-MEG3 knockdown suppressed NF-κB activation, and LncRNA-MEG3 was found to bind NF-κB components, promoting inflammatory cytokine production.
Conclusions:
- LncRNA-MEG3 acts as a positive regulator of the NF-κB pathway.
- Propofol exerts neuroprotection in LPS-triggered astrocyte injury by modulating the LncRNA-MEG3/NF-κB axis.
