VITAMIN D AND MAGNESIUM

Magnesium supplementation is recommended when taking vitamin D supplements. Magnesium helps activate vitamin D, which in turn helps regulate calcium and phosphate homeostasis to influence the growth and maintenance of bones. All the enzymes that metabolize vitamin D seem to require magnesium, which acts as a cofactor in the enzymatic reactions in the liver and kidneys. The dose of magnesium should be in the range of 250–500 mg/d, along with twice that dose of calcium. (Grant, 2020).

Reference:

(Grant, 2020). Grant WB, Lahore H, McDonnell SL, et al. Evidence that Vitamin D Supplementation Could Reduce Risk of Influenza and COVID-19 Infections and Deaths. Nutrients. 2020;12(4):988. Published 2020 Apr 2. doi:10.3390/nu12040988 https://pubmed.ncbi.nlm.nih.gov/32252338/

VITAMIN D AND GLUTATHIONE

Cells usually maintain a reducing environment and oxidative stress occurs when the cellular levels of ROS outbalance the antioxidants. If ROS levels become too high, the redox environment is driven out of homeostasis, resulting in the oxidation of proteins, DNA, and other cellular components. Mechanisms to maintain redox status, detoxify the ROS, and balance the thiol‐disulfide ratio are often triggered by the oxidation of thiol‐based redox switches. These pathways are often mediated by redox‐sensitive transcription factors, such as proteins with cysteine residues, which in conjunction with low‐molecular‐weight (LMW) thiols are scavengers of ROS such as H2O2. One of the most abundant LMW thiols is glutathione (GSH), a tripeptide (γ‐glutamyl cysteinyl glycine) that functions as a major endogenous antioxidant. GSH is the main cofactor for several enzymes that are responsible for detoxifying ROS. Homeostasis of the cellular redox environment is also maintained through the action of various ROS‐scavenging enzymes, including catalase, superoxide dismutase (SOD), glucose 6‐phosphate dehydrogenase, glutathione reductase (GR), and glutathione peroxidase. The regulation and function of these enzymes are dependent on cellular conditions and the physiological concentration of vitamin D.

Cytoplasmic conditions are more reduced and regulated, with only~10% of protein cysteines existing as disulfide bonds, compared to the cell surface environment. The extracellular environment lacks an effective redox‐homeostasis system, and therefore antioxidant enzymes and LMW thiols are crucial in maintaining the extracellular redox environment. Thiol‐disulfide balance affects binding of the SARS‐CoV‐2 virus with the host cell receptor; when the ACE2 receptor and S protein are in the reduced state, binding is thermodynamically unfavorable. Regulation of oxidative stress has a direct impact on COVID‐19 infection. This review on vitamin D elaborated on its role in maintaining the cellular redox status by regulating the expression of antioxidant enzymes and LMW thiols like GSH. In addition, it was observed that vitamin D affects the immune system; it modulates the innate and adaptive immune responses. Taken together, this review has provided a molecular‐level understanding of the role of this essential molecule in reducing the risks of viral infections including the COVID‐19. (Abdrabbo, 2021).

Reference:

(Abdrabbo, 2021).  Abdrabbo M, Birch CM, Brandt M, et al. Vitamin D and COVID-19: A review on the role of vitamin D in preventing and reducing the severity of COVID-19 infection. Protein Sci. 2021;30(11):2206-2220. doi:10.1002/pro.4190 Vitamin D and COVID‐19: A review on the role of vitamin D in preventing and reducing the severity of COVID‐19 infection – PMC (nih.gov)

VITAMIN D, VITAMIN K, AND MAGNESIUM

The COVID-19 pandemic is a current pandemic of high international interest, caused by the coronavirus strain SARS‑CoV‑2. Up to date, there is no treatment to decrease the virus-caused infection and mortality rates. More and more voices are being raised supporting the supplementation of Vitamin D3 to counter the pandemic outbreak with the correlated mortality rates as well as economic and social consequences. In a recently published review article, Sharma et al. (2020) have critically discussed the association of vitamin D with viral infections. A recent clinical study from Iran (n = 611) stated that there were no COVID-19 deaths in a hospital if serum 25(OH)D concentrations were higher than 41 ng/mL and patients were younger than 80. Russian hospitals observed that the likelihood to have severe COVID-19 increases by the factor of 5 if vitamin D is deficient. Similar observations have been made that there is a significant reduction in oxygen support for older clients when providing them with a relatively low daily dose of 1000 IU D3 OD, 150 mg magnesium OD, and 500 μg B12 OD upon admission.

Oral supplementation of D3 is the easiest means to prevent deficiencies. A frequent argument against supplementation of vitamin D3 is that an increased intake could lead to a vitamin D toxicity, also called hypervitaminosis D. This again can cause hypercalcemia, which is the buildup of calcium in the blood leading to vascular calcification, osteoporosis, and kidney stones. However, it has been reported that the reason for hypercalcemia rather lays in a vitamin K2 deficiency, as K2 activates the bone gamma-carboxyglutamic acid-containing protein (osteocalcin) through carboxylation. Activated osteocalcin deposits calcium in the bones, whereas non-activated osteocalcin inhibits calcium absorption by the bones. As the osteocalcin synthesis rate is increased by higher 25(OH)D serum levels, K2 is required as a natural antagonist.

Given that vitamin D3 is an immunoregulating hormone and can be considered safe when supplementing it together with K2, supplementation of magnesium (200–250 mg/day) should also be considered, as all enzymes that metabolize vitamin D3 seem to require magnesium. (Goddek, 2020).

Reference:

(Goddek, 2020). Goddek S. Vitamin D3 and K2 and their potential contribution to reducing the COVID-19 mortality rate. Int J Infect Dis. 2020 Oct;99:286-290. doi: 10.1016/j.ijid.2020.07.080. Epub 2020 Aug 6. PMID: 32768697; PMCID: PMC7406600. https://pubmed.ncbi.nlm.nih.gov/32768697/

QUERCETIN AS A ZINC IONOPHORE

Quercetin, a bioflavonoid polyphenol, has been shown to act as a zinc ionophore, enhancing entrance of zinc into cells to inhibit viral intracellular replication. It is also believed to block viruses from entering cells in the first place. An Oak Ridge National Labs/University of Tennessee study of many FDA-approved compounds presented supercomputer modeling results for inhibition by them of SARS-CoV-2 viral S-spike binding to cells. The study ranked quercetin as fifth out of 20 top performers. [33] Studies have shown quercetin also exhibiting anti-inflammatory properties, which could help mitigate the inflammatory response of cytokine and/or bradykinin storms provoked by COVID-19. A wide range of anti-viral/immunity benefits of quercetin have been identified, as well as other health benefits that may address some comorbidities of COVID-19 and some of its sequelae. (Margolin, 2021).

Reference:

(Margolin, 2021). Margolin L, Luchins J, Margolin D, Margolin M, Lefkowitz S. 20-Week Study of Clinical Outcomes of Over-the-Counter COVID-19 Prophylaxis and Treatment. J Evid Based Integr Med. 2021;26:2515690X211026193. doi:10.1177/2515690X211026193 https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8264737/

BROMELAIN, QUERCETIN, AND CURCUMIN

Due to its proteolytic action, bromelain is absorbed directly when administered orally, while it substantially promotes the absorption of curcumin and Quercetin enhancing its bioavailability and making this a perfect combination of immune-boosting nutraceuticals with synergistic anti-inflammatory and anticoagulant actions. (Panagiotis, et al., 2022).

Reference:

(Panagiotis, 2022). Kritis P, Karampela I, Kokoris S, Dalamaga M. The combination of bromelain and curcumin as an immune-boosting nutraceutical in the prevention of severe COVID-19. Metabol Open. 2020 Dec;8:100066. doi: 10.1016/j.metop.2020.100066. Epub 2020 Nov 13. PMID: 33205039; PMCID: PMC7661945.

BROMELAIN AND N-ACETYLCYSTEINE

The Combination of Bromelain and Acetylcysteine (BromAc) Synergistically Inactivates SARS-CoV-2. There is currently no suitable therapeutic treatment for early SARS-CoV-2 aimed at preventing disease progression. BromAc is under clinical development by the authors for mucinous cancers due to its ability to alter complex glycoprotein structures. The potential of BromAc on SARS-CoV-2 spike and envelope proteins stabilized by disulfide bonds was examined and found to induce the unfolding of recombinant spike and envelope proteins by reducing disulfide stabilizer bridges. BromAc also showed an inhibitory effecton wild-type and spike mutant SARS-CoV-2 by inactivation of its replication capacity in vitro. Hence, BromAc may be an effective therapeutic agent for early SARS-CoV-2infection, despite mutations, and even have potential as a prophylactic in people at high risk of infection.

Reference:

(Mekkawy, et.al., 2021). Research Gate: The Combination of Bromelain and Acetylcysteine (BromAc) Synergistically Inactivates SARS-CoV-2. Retrieved on01/17/23 online from https://www.researchgate.net/publication/349850528_The_Combination_of_Bromelain_and_Acetylcysteine_BromAc_Synergistically_Inactivates_SARS-CoV-2

GLUTATHIONE AND N-ACETYLCYSTEINE

Viral spike proteins play important roles in the viral entry process, facilitating attachment to cellular receptors and fusion of the viral envelope with the cell membrane. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein binds to the cellular receptor angiotensin converting enzyme-2 (ACE2) via its receptor-binding domain (RBD). The cysteine residue at position 488, consisting of a disulfide bridge with cysteine 480 is located in an important structural loop at ACE2-binding surface of RBD, and is highly conserved among SARS-related coronaviruses. We showed that the substitution of Cys-488 with alanine impaired pseudotyped SARS-CoV-2 infection, syncytium formation, and cell-cell fusion triggered by SARS-CoV-2 spike expression. Consistently, in vitro binding of RBD and ACE2, spike-mediated cell-cell fusion, and pseudotyped viral infection of VeroE6/TMPRSS2 cells were inhibited by the thiol-reactive compounds N-acetylcysteine (NAC) and a reduced form of glutathione (GSH). Furthermore, we demonstrated that the activity of variant spikes from the SARS-CoV-2 alpha and delta strains were also suppressed by NAC and GSH. Taken together, these data indicate that Cys-488 in spike RBD is required for SARS-CoV-2 spike functions and infectivity and could be a target of anti-SARS-CoV-2 therapeutics. (Murae, et al., 2022).

The antiviral activity of glutathione was demonstrated in a study of De Flora who showed that a 6-month preventive administration of N-acetylcysteine (NAC, glutathione precursor) significantly reduced the incidence of clinically apparent influenza and influenza-like episodes, especially in elderly high-risk individuals. In addition, pathophysiological conditions such as lung cell injury and inflammation in patients with severe ARDS were identified as the targets of NAC treatment. In particular, the deficiency of reduced glutathione in the alveolar fluid in acute respiratory distress syndrome (ARDS) patients was found to enhance lung cell injury by ROS/oxidative stress and inflammation, and this damage could be effectively prevented and treated by the administration of NAC. (Polonikov, 2020).

Systemic bioavailability of orally consumed glutathione is poor because the tripeptide is the substrate of proteases (peptidases) of the alimentary canal, and due to the absence of a specific carrier of glutathione at the level of cell membrane. (Allen, 2011), (Witschi, 1992).

Reference:

(Murae, et al., 2022). Biochemical and Biophysical Research Communications: The function of SARS-CoV-2 spike protein is impaired by disulfide-bond disruption with mutation at cysteine-488 and by thiol-reactive N-acetyl-cysteine and glutathione. Retrieved on 060722 online from https://www.sciencedirect.com/science/article/pii/S0006291X22001383?via%3Dihub#!

(Polonikov, 2020). Polonikov A. Endogenous Deficiency of Glutathione as the Most Likely Cause of Serious Manifestations and Death in COVID-19 Patients. ACS Infect Dis. 2020;6(7):1558-1562. doi:10.1021/acsinfecdis.0c00288

(Allen, 2011). Allen J, Bradley RD (September 2011). “Effects of oral glutathione supplementation on systemic oxidative stress biomarkers in human volunteers”. Journal of Alternative and Complementary Medicine. 17 (9): 827–33. doi:10.1089/acm.2010.0716. PMC 3162377. PMID 21875351.

(Witschi, 1992). Witschi A, Reddy S, Stofer B, Lauterburg BH (1992). “The systemic availability of oral glutathione”. European Journal of Clinical Pharmacology. 43 (6): 667–9. doi:10.1007/bf02284971. PMID 1362956. S2CID 27606314.

CURCUMIN AND PIPERINE

Orally administered curcumin with piperine could play a multifaceted role in the treatment of COVID-19. The anti-inflammatory and anti-thrombotic properties of curcumin could expedite the recovery of COVID-19 patients, and its antiviral, antibacterial, and antifungal properties could prevent superadded or secondary infections. Our results suggest that the use of orally administered curcumin with piperine as adjuvant therapy in COVID-19 treatment could substantially reduce morbidity and mortality, reduces treatment costs, and decrease logistical burden healthcare systems. Dose-escalating studies have indicated the safety of curcumin over 3 months. Hence, Curcumin can be a safe and natural therapeutic option to prevent Post-Covid thromboembolic events (Pawar, 2021).

Reference:

(Pawar, 2021). Oral Curcumin With Piperine as Adjuvant Therapy for the Treatment of COVID-19: A Randomized Clinical Trial. Retrieved on 061022 online from https://doi.org/10.3389/fphar.2021.669362.

CURCUMIN AND BROMELAIN

The coronavirus disease 2019 (COVID-19) pandemic is still ongoing, while no treatment has been proven effective. COVID-19 pathophysiology involves the activation of three main pathways: the inflammatory, the coagulation and the bradykinin cascades. Here, we highlight for the first time the joint potential therapeutic role of bromelain and curcumin, two well-known nutraceuticals, in the prevention of severe COVID-19. Bromelain (a cysteine protease isolated from the pineapple stem) and curcumin (a natural phenol found in turmeric) exert important immunomodulatory actions interfering in the crucial steps of COVID-19 pathophysiology. Their anti-inflammatory properties include inhibition of transcription factors and subsequent downregulation of proinflammatory mediators. They also present fibrinolytic and anticoagulant properties. Additionally, bromelain inhibits cyclooxygenase and modulates prostaglandins and thromboxane, affecting both inflammation and coagulation, and hydrolyzes bradykinin. Interestingly, curcumin has been shown in silico studies to prevent entry of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) into cells as well as viral replication, while a recent experimental study has demonstrated that bromelain may also inhibit viral entry into cells. Notably, bromelain substantially increases the absorption of curcumin after oral administration. To the best of our knowledge, this is the first report highlighting the significance of bromelain and, most importantly, the potential preventive value of the synergistic effects of bromelain and curcumin against severe COVID-19.

Clinical studies have demonstrated multiple beneficial effects of bromelain in trauma, ischemic injury, hypertension, atherosclerosis, inflammatory bowel disease, arthritis, and sinusitis as well as antibacterial and antifungal properties. Interestingly, a recent experimental study demonstrated that bromelain inhibits infection of VeroE6 cells by SARS-CoV-2 through blocking the virus binding and entry into cells via downregulation of ACE-2 and TMPRSS2 expression, and cleavage of the SARS-CoV-2 spike protein, presenting a novel promising therapeutic option that warrants further investigation. Due to its proteolytic action, bromelain is absorbed directly when administered orally, while it substantially promotes the absorption of curcumin enhancing its bioavailability and making this a perfect combination of immune-boosting nutraceuticals with synergistic anti-inflammatory and anticoagulant actions. (Kritis, 2020).

Reference:

(Kritis, 2020). Kritis P, Karampela I, Kokoris S, Dalamaga M. The combination of bromelain and curcumin as an immune-boosting nutraceutical in the prevention of severe COVID-19. Metabol Open. 2020;8:100066. doi:10.1016/j.metop.2020.100066

NIGELLA SATIVA AS A ZINC IONOPHORE

Having a range of bioactive components such as thymoquinone and nigellimine, black seed might offer a number of benefits to treat COVID-19 such as (i) blocking the entry of the virus into pneumocytes and (ii) providing ionophore for enhanced uptake of Zn2+ which in turn can enhance host immune response against SARS-CoV-2 as well as inhibit its replication by blocking the viral RdRp. (Rahman, 2020).

Reference:

(Rahman, 2020). Rahman MT. Potential benefits of combination of Nigella sativa and Zn supplements to treat COVID-19. J Herb Med. 2020;23:100382. doi:10.1016/j.hermed.2020.100382. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7313527/

GLUTATHIONE AND ZINC

To improve your glutathione, you need zinc. Another mechanism by which zinc acts as an antioxidant is by affecting the expression of glutamate-cysteine ligase, which is the rate-limiting enzyme of glutathione de novo synthesis. This has a two-fold effect of zinc to neutralize free radicals directly by glutathione or indirectly as a glutathione peroxidase cofactor. Ha showed that the administration of 100–150 mM of zinc in the cultured human retinal pigment epithelial cell line ARPE-19 cells upregulates the mRNA levels of glutamate-cysteine ligase via an nuclear factor erythroid 2 (NFE2)-related factor 2 (Nrf2)-dependent pathway. In this way, zinc modulates the total cellular glutathione concentration. (Marreiro, 2017).

Reference:

(Marreiro, 2017). Marreiro DD, Cruz KJ, Morais JB, Beserra JB, Severo JS, de Oliveira AR. Zinc and Oxidative Stress: Current Mechanisms. Antioxidants (Basel). 2017;6(2):24. Published 2017 Mar 29. doi:10.3390/antiox6020024

GLUTATHIONE AND SELENIUM

Selenium, a nutritionally essential micronutrient, is known for its antioxidant role in mitigating the effects caused by oxidative stress. Being an important component of various antioxidant selenoprotein enzymes like glutathione peroxidase (GPx) and thioredoxin reductase (TrxR), selenium plays an important role in combating oxidative stress caused due to excessive generation of ROS. The antioxidant enzymes like TrxR, apart from maintaining the redox status, also play a crucial role in regulating activities such as cell proliferation, cell death, and immune response activation. Further, selenoprotein P was found to enhance the activity and expression of glutathione peroxidase in the endothelial cell, thereby protecting the endothelial cells from oxidative damage.

Up-regulation of Nrf2 signaling by selenium has been shown to attenuate lung injury induced by sepsis. Activation of the Nrf2 pathway leads to the enhancement of glutathione synthesis and downregulation of the Nf-kβ pathway culminating in decreased lung injury. Conversely, the Nrf2 knockdown experiment had shown inhibition of glutathione synthesis and downregulation of the NF-κB pathway. NF-κB transcription factor is the final common step of the regulation for the expression of inflammatory cytokines. The modulation of its activity has been the target for many anti-inflammatory drugs. The effect of selenium on Nrf2 and NF-κB signaling culminates in the regulation of inflammatory cytokines. Selenium supplementations have been found to decrease the production of inflammatory cytokines like IL-6 in cell line studies, animal models, and human studies. (Tomo, 2021).

Reference:

(Tomo, 2021). Tomo S, Saikiran G, Banerjee M, Paul S. Selenium to selenoproteins – role in COVID-19. EXCLI J. 2021;20:781-791. Published 2021 Apr 16. doi:10.17179/excli2021-3530

GLUTATHIONE AND ASTAXANTHIN

Glutathione and Astaxanthin work synergistically and collectively to form a barrier against pathogens. In this study investigated the effects of 6 mg/day of astaxanthin supplementation on markers of oxidative stress and substrate metabolism during a graded exercise test in active young men. A double-blind, randomized, counterbalanced, cross-over design was used. Fourteen men (age = 23 ± 2 years) supplemented with 6 mg/day of astaxanthin and a placebo for 4 weeks, with a 1 week washout period between treatments. Following each supplementation period, a fasting blood sample was obtained to measure markers of oxidative stress: glutathione, hydrogen peroxide, advanced oxidation protein products, and malondialdehyde. Participants also completed a graded exercise test after each treatment to determine substrate utilization during exercise at increasing levels of intensity. Glutathione was ∼7% higher following astaxanthin compared with placebo (1,233 ± 133 vs. 1,156 ± 185 μM, respectively; p = .02, d = 0.48). Astaxanthin supplementation of 6 mg/day for 4 weeks increased whole blood levels of the antioxidant glutathione in active young men but did not affect oxidative stress markers or substrate utilization during exercise. Astaxanthin appears to be an effective agent to increase endogenous antioxidant status (McAllister, 2022).

Reference:

(McAllister, 2022). McAllister MJ, Mettler JA, Patek K, Butawan M, Bloomer RJ. Astaxanthin Supplementation Increases Glutathione Concentrations but Does Not Impact Fat Oxidation During Exercise in Active Young Men. Int J Sport Nutr Exerc Metab. 2022 Jan 1;32(1):8-15. doi: 10.1123/ijsnem.2021-0138. Epub 2021 Oct 5. PMID: 34611051. https://pubmed.ncbi.nlm.nih.gov/34611051/