Ophiopogon Polysaccharide Liposome Regulated the Immune Activity of Kupffer Cell through miR-4796
Abstract
:1. Introduction
2. Results
2.1. The Expression of miR-4796 after Transfected miR-4796 Mimic or Inhibitor
2.2. The Effect of OPL on the Secretion of NO and iNOS
2.3. The Effect of OPL on Cell Migration
2.4. The Effect of OPL on the Phagocytosis of KCs for FITC-Dextran by Fluorescence Staining
2.5. The Effect of OPL on the Phagocytosis of KCs for FITC-Dextran by Flow Cytometry
2.6. The Effect of OPL on the Phagocytosis of KCs for FITC-OVA
2.7. The Effect of OPL on the Cell Apoptosis
2.8. The Effect of OPL on the Expression of CD14 and MHCII
2.9. The Effect of OPL on the Secretion of ROS
2.10. The Effect of OPL on the mRNA and Protein Expression of TLR4, MyD88, IKKβ and NF-κB
3. Discussion
4. Materials and Methods
4.1. Materials
4.1.1. Cell
4.1.2. Reagent
4.2. Methods
4.2.1. The Preparation of Ophiopogon Polysaccharide Liposomes
4.2.2. Effects of miR-4796 on the Biological Function of KCs Regulated by OPL
The Expression of miR-4796 after Transfected with miR-4796 Mimic or Inhibitor by Real-Time PCR Method
miR-4796 Transfection and Treatment of OPL
Measurement of NO
Measurement of iNOS
Cell Scratch Test
Measurement of Phagocytosis of KCs on FITC-Dextran by Fluorescence Staining and Flow Cytometry
Measurement of Phagocytosis of KCs on FITC-OVA by Fluorescence Staining
Measurement of Apoptosis by Hoechst 33,258 Staining
Measuring the Expression Levels of CD14 and MHC II by Flow Cytometry
Measurement of ROS by Fluorescence Staining
4.2.3. The Function of miR-4796 in TLR4-NF-κB Signaling Pathway Regulated by OPL
Measuring the mRNA Expression of TLR4, IKKβ, MyD88 and NF-κB by Real-Time PCR
Measuring the Protein Expression of TLR4, IKKβ, MyD88 and NF-κB by Western Blot
4.2.4. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Cui, J.; Pan, X.; Duan, X.; Ke, L.; Song, X.; Zhang, W.; Ma, W.; Liu, Y.; Fan, Y. Ophiopogon Polysaccharide Liposome Regulated the Immune Activity of Kupffer Cell through miR-4796. Int. J. Mol. Sci. 2022, 23, 14659. https://doi.org/10.3390/ijms232314659
Cui J, Pan X, Duan X, Ke L, Song X, Zhang W, Ma W, Liu Y, Fan Y. Ophiopogon Polysaccharide Liposome Regulated the Immune Activity of Kupffer Cell through miR-4796. International Journal of Molecular Sciences. 2022; 23(23):14659. https://doi.org/10.3390/ijms232314659
Chicago/Turabian StyleCui, Jing, Xingxue Pan, Xueqin Duan, Liting Ke, Xiaoping Song, Weimin Zhang, Wuren Ma, Yingqiu Liu, and Yunpeng Fan. 2022. "Ophiopogon Polysaccharide Liposome Regulated the Immune Activity of Kupffer Cell through miR-4796" International Journal of Molecular Sciences 23, no. 23: 14659. https://doi.org/10.3390/ijms232314659