Polyhexamethylene Guanidine Phosphate Damages Tight Junctions and the F-Actin Architecture by Activating Calpain-1 via the P2RX7/Ca2+ Signaling Pathway
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemicals and Reagents
2.2. Cell Culture
2.3. Cell Viability Assay
2.4. Immunofluorescence
2.5. Determination of Intracellular Ca2+ Level
2.6. Measurement of Calpain Activity
2.7. ATP Assay
2.8. Western Blotting
2.9. Statistical Analysis
3. Results
3.1. Effect of PHMG-p on TJ Proteins and the F-Actin Architecture in BEAS-2B Cells
3.2. Role of Calpain in Impairment of TJ Proteins and F-Actin Architecture by PHMG-p
3.3. PHMG-p Induces Intracellular Ca2+ Influx via P2RX7
3.4. PHMG-p Induces Calpain-1 Activity and Calpain-1-Dependent Epithelial Barrier Dysfunction via P2RX7/Ca2+
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
AJs | adherens junctions |
DMEM/F-12 | Dulbecco’s modified Eagle’s medium/Ham’s F-12 Nutrient Mixture |
eATP | extracellular ATP |
LDH | lactate dehydrogenase |
MTT | 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide |
PHMG-p | polyhexamethylene guanidine phosphate |
P2RX7 | P2X purinoreceptor 7 |
TJs | tight junctions |
ZO | zonula occludens |
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Jin, S.W.; Lee, G.H.; Pham, H.T.; Choi, J.H.; Jeong, H.G. Polyhexamethylene Guanidine Phosphate Damages Tight Junctions and the F-Actin Architecture by Activating Calpain-1 via the P2RX7/Ca2+ Signaling Pathway. Cells 2020, 9, 59. https://doi.org/10.3390/cells9010059
Jin SW, Lee GH, Pham HT, Choi JH, Jeong HG. Polyhexamethylene Guanidine Phosphate Damages Tight Junctions and the F-Actin Architecture by Activating Calpain-1 via the P2RX7/Ca2+ Signaling Pathway. Cells. 2020; 9(1):59. https://doi.org/10.3390/cells9010059
Chicago/Turabian StyleJin, Sun Woo, Gi Ho Lee, Hoa Thi Pham, Jae Ho Choi, and Hye Gwang Jeong. 2020. "Polyhexamethylene Guanidine Phosphate Damages Tight Junctions and the F-Actin Architecture by Activating Calpain-1 via the P2RX7/Ca2+ Signaling Pathway" Cells 9, no. 1: 59. https://doi.org/10.3390/cells9010059