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Updated: Dec 30, 2025
Isolation of Murine Peritoneal Macrophages to Carry Out Gene Expression Analysis Upon Toll-like Receptors Stimulation
Published on: April 29, 2015
Analysis of microRNA expression profiles in porcine PBMCs after LPS stimulation
Jing Zhang1, Xin Xu1, Xingfa Huang2
1Hubei Key Laboratory of Animal Nutrition and Feed Science, Hubei Collaborative Innovation Center for Animal Nutrition and Feed Safety, Wuhan Polytechnic University, PR China.
Abstract:
In the present study, we used microRNA (miRNA) sequencing to discover and explore the expression profiles of known and novel miRNAs in 1000 ng/ml LPS stimulated for 8 h vis-à-vis non-stimulated (i.e. control) PBMCs isolated from the blood of healthy pigs. A total of 291 known miRNAs were bio-computationally identified in porcine PBMCs, and 228 novel miRNAs (not enlisted in the swine mirBase) were identified. Among these miRNAs, ssc-miR-148a-3p, ssc-let-7g, ssc-let-7f, 3_8760, ssc-miR-26a, ssc-miR-451, ssc-miR-21, ssc-miR-30d, ssc-miR-99a and ssc-miR-103 were the top 10 most abundant miRNAs in porcine PBMCs. Through miRNA differential analysis combined with quantitative PCR, we found the expressions of ssc-miR-122, ssc-miR-129b, ssc-miR-17-5p and ssc-miR-152 were significantly changed in porcine PBMCs after LPS stimulation. Furthermore, targets prediction and function analysis indicated a significant enrichment in gene ontology functional categories related to diseases, immunity and inflammation. In conclusion, this study on profiling of miRNAs expressed in LPS-stimulated PBMCs provides an important reference point for future studies on regulatory roles of miRNAs in porcine immune system.
Insights
This study profiled microRNAs (miRNAs) in pig immune cells, identifying known and novel miRNAs. LPS stimulation significantly altered specific miRNA expressions, offering insights into swine immunity regulation.
Area of Science:
- Veterinary Immunology
- Molecular Biology
- Genomics
Background:
- MicroRNAs (miRNAs) play crucial roles in regulating gene expression.
- Understanding miRNA profiles in pigs is vital for advancing swine health and disease management.
- Lipopolysaccharide (LPS) stimulation is a common method to mimic bacterial infection and study immune responses.
Purpose of the Study:
- To discover and characterize known and novel microRNAs (miRNAs) in porcine peripheral blood mononuclear cells (PBMCs).
- To investigate the differential expression of miRNAs in PBMCs upon lipopolysaccharide (LPS) stimulation.
- To predict the functional roles of identified miRNAs in the context of swine immunity and inflammation.
Main Methods:
- MicroRNA (miRNA) sequencing was performed on LPS-stimulated and non-stimulated porcine PBMCs.
- Bio-computational analysis was used to identify known and novel miRNAs.
- Differential expression analysis and quantitative PCR (qPCR) were employed to validate miRNA expression changes.
- Target prediction and gene ontology (GO) analysis were conducted to infer miRNA functions.
Main Results:
- A comprehensive profile of 291 known and 228 novel miRNAs in porcine PBMCs was established.
- The top 10 most abundant miRNAs were identified, including ssc-miR-148a-3p and ssc-let-7 family members.
- LPS stimulation significantly altered the expression of specific miRNAs, such as ssc-miR-122, ssc-miR-129b, ssc-miR-17-5p, and ssc-miR-152.
- Functional analysis revealed enrichment in pathways related to disease, immunity, and inflammation.
Conclusions:
- This study provides a valuable miRNA expression profile for porcine PBMCs, particularly under LPS stimulation.
- The identified differentially expressed miRNAs are potential regulators of the immune response in pigs.
- These findings serve as a foundation for future research into miRNA-mediated regulation of the porcine immune system.
