Related Experiment Video
Updated: Aug 22, 2025
Protocol for the Differentiation of Human Induced Pluripotent Stem Cells into Mixed Cultures of Neurons and Glia for Neurotoxicity Testing
Published on: June 9, 2017
Blockage of KHSRP-NLRP3 by MCC950 Can Reverse the Effect of Manganese-Induced Neuroinflammation in N2a Cells and Rat
Sharad Singh1, Ibrahim Ahmed Shaikh2, Sunil S More1
1School of Basic and Applied Sciences, Dayananda Sagar University, Bangalore 560111, Karnataka, India.
Abstract:
Manganese neurotoxicity has been reported to cause a neurodegenerative disease known as parkinsonism. Previous reports have shown that the expression of the KH-type splicing regulatory protein (KHSRP), a nucleic acid-binding protein, and NLRP3 is increased upon Mn exposure. However, the relation between these two during Mn toxicity has not been fully deduced. The mouse neuroblastoma (N2a) and SD rats are treated with LPS and MnCl2 to evaluate the expression of KHSRP and NLRP3. Further, the effect of the NLRP3 inhibitor MCC950 is checked on the expression of NLRP3, KHSRP and pro-inflammatory markers (TNFα, IL-18 and IL-1β) as well as the caspase-1 enzyme. Our results demonstrated an increment in NLRP3 and KHSRP expression post-MnCl2 exposure in N2a cells and rat brain, while on the other hand with LPS exposure only NLRP3 expression levels were elevated and KHSRP was found to be unaffected. An increased expression of KHSRP, NLRP3, pro-inflammatory markers and the caspase-1 enzyme was observed to be inhibited with MCC950 treatment in MnCl2-exposed cells and rats. Manganese exposure induces NLRP3 and KHSRP expression to induce neuroinflammation, suggesting a correlation between both which functions in toxicity-related pathways. Furthermore, MCC950 treatment reversed the role of KHSRP from anti-inflammatory to pro-inflammatory.
Insights
Manganese exposure increases KH-type splicing regulatory protein (KHSRP) and NLRP3, driving neuroinflammation. Inhibiting NLRP3 with MCC950 reduces these effects, suggesting a link between KHSRP and NLRP3 in manganese toxicity.
Area of Science:
- Neuroscience
- Toxicology
- Molecular Biology
Background:
- Manganese neurotoxicity can lead to parkinsonism.
- Manganese exposure elevates KH-type splicing regulatory protein (KHSRP) and NLRP3 expression.
- The precise relationship between KHSRP and NLRP3 in manganese toxicity is not fully understood.
Purpose of the Study:
- To investigate the correlation between KHSRP and NLRP3 expression during manganese-induced neurotoxicity.
- To evaluate the therapeutic potential of the NLRP3 inhibitor MCC950 in manganese toxicity.
Main Methods:
- Mouse neuroblastoma (N2a) cells and SD rats were exposed to lipopolysaccharide (LPS) and manganese chloride (MnCl2).
- Expression levels of KHSRP, NLRP3, pro-inflammatory markers (TNFα, IL-18, IL-1β), and caspase-1 were analyzed.
- The effects of MCC950 treatment on these markers were assessed in MnCl2-exposed models.
Main Results:
- MnCl2 exposure increased both NLRP3 and KHSRP expression in N2a cells and rat brains.
- LPS exposure elevated NLRP3 but did not affect KHSRP levels.
- MCC950 treatment inhibited MnCl2-induced increases in KHSRP, NLRP3, pro-inflammatory markers, and caspase-1 activity.
- MCC950 treatment shifted KHSRP's role from anti-inflammatory to pro-inflammatory.
Conclusions:
- Manganese exposure induces neuroinflammation by upregulating both NLRP3 and KHSRP, indicating a functional correlation.
- MCC950 effectively mitigates manganese-induced neuroinflammation and associated molecular changes.
- KHSRP's inflammatory role can be modulated by NLRP3 inhibition.
