MicroRNA-146a-5p Negatively Regulates Pro-Inflammatory Cytokine Secretion and Cell Activation in Lipopolysaccharide Stimulated Human Hepatic Stellate Cells through Inhibition of Toll-Like Receptor 4 Signaling Pathways
Abstract
:1. Introduction
2. Results
2.1. Expression of Toll-Like Receptor 4 (TLR4), α-Smooth Muscle Actin (α-SMA) and miR-146a-5p in LX2 Cells Stimulated with Lipopolysaccharide (LPS)
2.2. Overexpression of miR-146a-5p Reduced LPS Induced Pro-Inflammatory Cytokines Secretion
2.3. miR-146a-5p Inhibited Cytokines Secretion by Down-Regulating the Downstream Genes of TLR4 Signaling Way
2.4. Knockdown of IL-1 Receptor-Associated Kinase 1 (IRAK1) and TNF Receptor Associated Factor-6 (TRAF6) Suppressed LPS Induced Pro-Inflammatory Cytokines Production
2.5. Overexpression of miR-146a-5p Attenuated LPS Induced c-Jun N-Terminal Kinase (JNK) Activation and α-SMA Expression
2.6. Knockdown of TRAF6 but Not IRAK1 Inhibited LPS Induced JNK Activation and α-SMA Expression
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. miRNA Inhibitor or Mimic Transfection
4.3. Quantitative Real-Time PCR
4.4. RNA Interference
4.5. ELISA
4.6. Western Blot
4.7 Statistical Analyses
5. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Chen, Y.; Zeng, Z.; Shen, X.; Wu, Z.; Dong, Y.; Cheng, J.C.-H. MicroRNA-146a-5p Negatively Regulates Pro-Inflammatory Cytokine Secretion and Cell Activation in Lipopolysaccharide Stimulated Human Hepatic Stellate Cells through Inhibition of Toll-Like Receptor 4 Signaling Pathways. Int. J. Mol. Sci. 2016, 17, 1076. https://doi.org/10.3390/ijms17071076
Chen Y, Zeng Z, Shen X, Wu Z, Dong Y, Cheng JC-H. MicroRNA-146a-5p Negatively Regulates Pro-Inflammatory Cytokine Secretion and Cell Activation in Lipopolysaccharide Stimulated Human Hepatic Stellate Cells through Inhibition of Toll-Like Receptor 4 Signaling Pathways. International Journal of Molecular Sciences. 2016; 17(7):1076. https://doi.org/10.3390/ijms17071076
Chicago/Turabian StyleChen, Yuhan, Zhaochong Zeng, Xiaoyun Shen, Zhifeng Wu, Yinying Dong, and Jason Chia-Hsien Cheng. 2016. "MicroRNA-146a-5p Negatively Regulates Pro-Inflammatory Cytokine Secretion and Cell Activation in Lipopolysaccharide Stimulated Human Hepatic Stellate Cells through Inhibition of Toll-Like Receptor 4 Signaling Pathways" International Journal of Molecular Sciences 17, no. 7: 1076. https://doi.org/10.3390/ijms17071076