N-acetylcysteine Reduces Inflammasome Activation Induced by SARS-CoV-2 Proteins In Vitro
Abstract
:1. Introduction
2. Results
2.1. Inflammasome Components Are Overexpressed in Airway Tissue of COVID-19 Critically Ill Patients
2.2. The Combination of SARS-CoV-2 Proteins S, E, and M Promotes the Release of Inflammatory Proteins in Human Macrophages and Bronchial Epithelial Cells
2.3. N-acetylcysteine Inhibits SARS-CoV-2 Mosaic (S-E-M) Inflammation in a Concentration and Time-Dependent Fashion in Macrophages and Bronchial Epithelial Cells
2.4. N-acetylcysteine Inhibits the SARS-CoV-2 Mosaic (S-E-M)-Induced Inflammasome Activation and Intracellular Nf-κB and ERK1/2 Inflammatory Pathways
2.5. Effects of Mosaic (S-E-M) SARS-CoV-2 Protein on Oxidative Stress, Thiols, and Glutathione
3. Discussion
4. Materials and Methods
4.1. Patients
4.2. Real-Time qRT-PCR and Gene Silencing
4.3. Immunohistochemical and Immunofluorescence Studies
4.4. Isolation and Culture of Human Bronchial Epithelial Cells and Mononuclear Cells
4.5. In Vitro Stimulations and Conditions
4.6. Western Blot
4.7. CM-H2DCFDA Fluorescence Measurement of Reactive Oxygen Species
4.8. CM-FDA Fluorescence Measurement of Thiols
4.9. Measure of GSH
4.10. Statistical Analysis
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Variable | Control Cohort | Covid-19 (B.1) | Covid-19 (B.1.1.7 VOC) |
---|---|---|---|
Number of patients | 32 | 31 | 20 |
Age (years) [IQR] | 65.2 [58.5–74] | 67.7 [60.5–75.5] | 80 [69.7–91.7] *, # |
Male (%) | 26 (81.2) | 18 (58.1) | 9 (45) |
Exitus, n (yes %) | 19 (59.4) | 17 (54.8) | 20 (100) *, # |
28-day mortality, n (yes %) | 13 (40.6) | 4 (12.9)* | 19 (95) *, # |
Smoker (yes %) | 17 (53.1) | 11 (35.5) | 8 (40) |
Diabetes (yes %) | 6 (18.7) | 6 (19.3) | 7 (35) |
COPD (yes %) | 5 (15.6) | 0 (0) | 2 (10) |
Arterial hypertension (yes %) | 15 (46.8) | 14 (45.1) | 17 (85) *, # |
Dyslipemia (yes %) | 11 (34.4) | 11 (35.4) | 11 (55) |
BMI (kg/m2) [IQR] | 26.8 [21.4–40.9] | 32 [28.8–33.3] | 30.2 [25.7–33.7] |
Lymphocytes (10 * 9/L) [IQR] | 0.95 [0.5–1.3] | 1.05 [0.5–1.27] | 0.67 [0.3–0.8] |
Neutrophils (10 * 9/L) [IQR] | 9.4 [6.3–12.8] | 8.7 [5.9–11.2] | 14.2 [6.5–20.5] |
CRP (mg/L) [IQR] | 8.4 [0.8–15.4] | 5.8 [0.3–9.2] | 14.05 [6.7–19.8] *, # |
LDH (U/L) [IQR] | 702 [630–774] | 584.3 [496–666] | 951 [692–1111] *,# |
D-dimer (ng/mL) [IQR] | 332 [263–425] | 1593 [536–1929] | 2797 [251–1473] |
Procalcitonin (ng/mL) [IQR] | 3.49 [0.3–1.9] | 0.43 [0.07–1.25] * | 7.45 [0.08–7.6] |
pO2 (mmHg) [IQR] | 148.1 [98.1–188.5] | 138.1 [80.5–174.8] | 92.2 [53–115] *, # |
pCO2 (mmHg) [IQR] | 42.9 [37–52] | 48.9 [42.2–52.7] | 45.6 [41–52] |
PaFiO2 [IQR] | 279.8 [214–366] | 240.1 [193–258] | 220.2 [166–274] |
HCO3 (mmoL/L) [IQR] | 24.8 [20.4–27.9] | 31.6 [27.8–33.6] * | 27.9 [24.8–31.3] |
Ferritin (µg/L) [IQR] | 855 [240–1305] | 930.9 [399–1443] | 1045 [389–1448] |
ALT (U/L) [IQR] | 41.8 [10.7–60] | 73.3 [32–104] | 61.8 [23–70] |
Systemic corticosteroids, n (%) | 17 (53.1) | 25 (80.6) * | 18 (90) * |
NRLP3 mRNA Expression vs. | Spearman ρ | p Value |
---|---|---|
Age (years) | 0.4065 | 0.0092 * |
Leukocytes (109/L) | 0.2122 | 0.194 |
Lymphocytes (109/L) | −0.1716 | 0.2962 |
Neutrophils (109/L) | 0.2370 | 0.1464 |
Monocytes (109/L) | 0.04033 | 0.8074 |
CRP (mg/dL) | 0.5374 | 0.0003 * |
LDH (U/L) | 0.5193 | 0.0016 * |
Ferritin (µg/L) | 0.3262 | 0.0597 |
ALT (U/L) | −0.04604 | 0.7867 |
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Milara, J.; Martínez-Expósito, F.; Montero, P.; Roger, I.; Bayarri, M.A.; Ribera, P.; Oishi-Konari, M.N.; Alba-García, J.R.; Zapater, E.; Cortijo, J. N-acetylcysteine Reduces Inflammasome Activation Induced by SARS-CoV-2 Proteins In Vitro. Int. J. Mol. Sci. 2022, 23, 14518. https://doi.org/10.3390/ijms232314518
Milara J, Martínez-Expósito F, Montero P, Roger I, Bayarri MA, Ribera P, Oishi-Konari MN, Alba-García JR, Zapater E, Cortijo J. N-acetylcysteine Reduces Inflammasome Activation Induced by SARS-CoV-2 Proteins In Vitro. International Journal of Molecular Sciences. 2022; 23(23):14518. https://doi.org/10.3390/ijms232314518
Chicago/Turabian StyleMilara, Javier, Fernando Martínez-Expósito, Paula Montero, Inés Roger, Maria Amparo Bayarri, Pilar Ribera, Miriam Natsuki Oishi-Konari, Jose Ramón Alba-García, Enrique Zapater, and Julio Cortijo. 2022. "N-acetylcysteine Reduces Inflammasome Activation Induced by SARS-CoV-2 Proteins In Vitro" International Journal of Molecular Sciences 23, no. 23: 14518. https://doi.org/10.3390/ijms232314518