Inflammation NF-kB

Inflammation & NF-kB

 

Wow, what a mess the world has become regarding the massive problems associated with Inflammation, now including a viral infection that causes inflammation of the airways. It is of concern that virtually all the globes financial focus and support hinges around the new patentable technologies that attempt to address ever-evolving and mutating components that now appear to be here to stay in some form or another, regardless of using this year’s vaccine or next year’s variant. We believe that it is essential to focus on inflammation in its entirety regardless of the cause be it acute or chronic, and irrespective of current challenges or variants that may emerge into the future.

 

 

 

 

Ultimately, we seem to be experiencing a pandemic of inflammation which is already rampant throughout the world due to an ever-increasing accumulation of reasons be it food stuffs, farming methods, contaminants or environment. Virtually all the consequences thereafter result from not managing that inflammation, which is primarily a biological function that is powered by ATP. The amount of ATP the mitochondria are able to produce directly effects the amount of work that can be done in a cell and ultimately enabling cellular homeostasis.

 

 

Inflammation

 
Inflammation is a generic response of body tissue following damage to our cells or detecting a potentially damaging effector. Both infectious and non-infectious agents, cell damage and Endoplasmic Reticulum Stress (Unfolded Protein Response), can activate an inflammatory response. The response is generic because it doesn’t distinguish between the possible causes. The inflammatory response removes harmful stimuli and initiates a healing process that involves both the immune and circulatory systems however, it is a balancing act that can go awry. Molecular hydrogen acts against inflammation by reducing pro-inflammatory cytokine production, decreasing oxidative stress, and increasing the production of ATP.

 

Inflammation as a response to injury

 

 

First, blood vessels adjacent to the injury undergo vasoconstriction (they somehow close up). Thus at a local level blood flow is reduced, as if the body is trying to limit potential blood loss at the injury site before further evaluation is conducted. Indeed, this first response is only transient and soon after, the blood vessel vasodilates, increasing blood flow to the area. This increase allows more nutrients and more oxygen to be brought to the injured area to facilitate repairs. It also brings more immune cells to the site. These cells will participate and promote the inflammatory response that is taking place around the injury.

 

 

Immune Cells and Inflammation

 

 

The cellular aspect of inflammation is complex, and research reveals more of this complexity every day. The main cellular effectors of the immune response are lymphocytes (T-cells, B-cells, and NK cells), neutrophils, and monocytes/macrophages. NK cells, neutrophils, and monocytes/macrophages drive the innate immune response. It doesn’t require any “learning.” Whereas the adaptative immune response caused by T-cells and B-cells does require “learning.” While B-cells are also responsible for anti-body production, all these cellular effectors of immunity produce molecules that modulate the immune response called cytokines. The primary producers of cytokines are lymphocytes and macrophages. Some are proinflammatory, others are anti-inflammatory, but generalising a particular cytokine’s effect is virtually impossible due to the system’s complexity. Regardless, deregulated cytokine secretion is implicated in several disease states ranging from chronic inflammation to allergy. The activation of immune cells sees the metabolic pathway used to produce energy change in these cells from the more energy-efficient oxidative phosphorylation, to the more rapid but less energy-efficient glycolysis. This switch also occurs in anoxic conditions and rapidly multiplying cells such as cancer cells. The reverse metabolic change is fundamental to the resolution of inflammation. This change in the metabolic pathway is central to the development of inflammation.

 

Macrophages develop from monocytes1 and are at the forefront of the initial immune response. Macrophages are said to have two extreme phenotypes, M1 and M2. The M1 phenotype promote inflammation and directly deals with intruders by phagocytose while the M2 phenotype macrophage contribute to tissue repair and promote healing. Although there are M1-like and M2-like macrophages, there are also intermediate versions, and macrophages can transition from one type to the other1. That said, they have both a critical role to play during inflammation.

 

 

Following the detection of a potential intruder, inflammatory responses can be triggered by macrophages, including other monocyte-derived phagocytic cells and MAST cells. Mast cells are long-lived tissue-resident cells involved in the host defense against parasitic infection and allergic reactions. Once activated, they can release a wide variety of inflammatory mediators such as histamine, Interleukin 4 (IL-4), Interleukin 6 (IL-6), Interleukin 13 (IL-13), Tumor Necrosis Factor Alpha (TNF-alpha), and various other molecules. The latter will increase blood vessels’ permeability and attract more immune cells to the site, including M1 macrophage. Resident macrophages are generally of the M2-like type2. Their response to injury or pathogen detection will be to attract and facilitate the migration of M1 types to the site from the blood vessels. Although M2 macrophages are associated with healing and tissue repair, M1 macrophages produce proinflammatory cytokines, phagocyte bacteria, release Nitric Oxide (NO) and reactive oxygen species (ROS) to kill bacteria and viruses. The latter is essential ROS induce oxidative stress both for the intruders and the body cells. Generally, eucaryote cells such as mammalian cells are better at resisting oxidative stress than, let’s say, bacteria. Nonetheless, there are limits, and if the balance between M1 and M2 macrophage work is not maintained, things will go awry. Generally, it is not the case, and the body will get rid of the problem and heal. However, repair means energy, and the energy centres of eucaryote cells are the mitochondria.

 

Mitochondria use oxygen to produce energy for the cell; the more energy is needed, the more ROS are produced by the mitochondria, especially if insufficient oxygen is available, increasing the cell’s exposure to oxidative stress. ROS production is further exacerbated by the metabolic switch from oxidative phosphorylation to glycolysis. Interestingly, there is an interconnection between Reactive Oxygen Species production and inflammation. ROS can trigger inflammation3, and inflammation results in ROS production, enabling the possibility of self-sustaining inflammation. The nuclear factor kappa B protein family lay at the heart of inflammation control.

The Heart of Inflammation – NF-κB

NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) is a protein complex that controls transcription of DNA, cytokine production, and cell survival. NF-κB is found in almost all animal cell types (including insects and cnidarian)4. NF-κB family members include NF-kB1, NF-kB2, RelA, RelB, and c-Rel in mammalian4. It is involved in cellular responses to stimuli such as stress, cytokines, free radicals, heavy metals, ultraviolet irradiation, oxidised LDL, and bacterial or viral antigens. In addition, NF-κB plays a crucial role in regulating the immune response to infection. Thus, incorrect regulation of NF-κB is linked to cancer5, inflammatory and autoimmune diseases6,7, septic shock7, viral infection8, and improper immune development.
 

NF-κB protein complexes are sequestered in the cell cytoplasm until activated. Once activated, they translocate in the cell’s nucleus and modify gene expression9 (note that NF-κB has also been found in the Mitochondria, however little is known about its action there10). NF-κB complexes can be activated through the canonical or classical pathway, the noncanonical pathway, and the atypical pathway (though DNA damage). Each pathway involving different/same members of the NF-kB family. Indeed, the formal separation in pathways is for practical reasons. For example, the activation of the noncanonical pathway can lead to the activation of some or all of the classical pathways. Nonetheless, oxidative stress can modulate NF-kB pathways11, sometimes activating, sometimes inhibiting NF-kB at a high or specific gene level, as a function of molecular context and cell types. NF-κB is a central mediator of inflammation, promoting cytokine expression, and its deregulation can lead to cytokine storms6.  

Molecular Hydrogen modulates the action of NF-kB, decreasing inflammation and preventing apoptosis in normal cells while promoting it in cancerous cells. Molecular hydrogen also reverses the energy metabolic pathway switch back from glycolysis to oxidative phosphorilation29 promoting the resolution of the inflammation.

NF-kB and Inflammatory Diseases


NF-kB dysregulation in association with elevated levels of ROS is implicated in the pathogenesis of many inflammatory diseases6 such as type I and II diabetes, Chronic obstructive pulmonary disease, asthma, Rheumatoid arthritis, Inflammatory bowel disease, multiple sclerosis, atherosclerosis, and Ankylosing Spondylitis6,12,13. But also in the parthenogenesis of a lot of infectious diseases. In this regards, COVID 19 need a special mention. Recent work8 has shown that the ORF3a, M, ORF7a, and N proteins of SARS‑CoV‑2 virus were NF‑κB activators. For example, the ORF7a protein induced the NF‑κB dependent proinflammatory cytokines including IL‑1α, IL‑1β, IL‑6, IL‑8, IL‑10, TNF‑α. Davies et al (2021)14 have also noted that NF-κB is elevated in a dose-dependent manner in response to covid.

Molecular Hydrogen has already proved beneficial to sufferers of acute exacerbation of chronic obstructive pulmonary disease15, diabetes Mellitus type I16 (animal model) and type II17,18, rheumatoid arthritis19,20, atherosclerosis21,22, asthma23, and an animal model of multiple sclerosis or Inflammatory bowel disease23 through its potent anti-oxidant effect and well documented dampening effect on NF-κB 24.

Given this and given the positive outcome of clinical trial on COVID 19 patient25, Oxy-hydrogen inhalation appear to provide with a high potential therapeutical adjuvant for the treatment and prevention of severe COVID-19.

The Beneficial Action of Molecular Hydrogen


Molecular Hydrogen has proven to act as a powerful antioxidant in biological systems both directly and indirectly.

  • Directly, by scavenging (neutralising) some ROS, such as the infamous hydroxyl radical26
  • Indirectly, by boosting the production of enzyme parts of the natural antioxidant system such as catalase and superoxide dismutase.

Molecular Hydrogen also appears to potentiate the normal functioning of the mitochondria by restoring the use of oxidative phosphorylation, but the action of Molecular Hydrogen concerning inflammation doesn’t stop there.

Molecular Hydrogen also seems to decrease the relative number of M1 compared to the healing-promoting M2 macrophage27,28, probably by reversing the metabolic switch or promoting its reversal. In doing so, Molecular Hydrogen avoids an imbalance between M1 and M2 macrophages, helping to bring about healing by decreasing inflammation.

Finally, Molecular Hydrogen seems to be able to downregulate NF-κB 24 providing us with a new therapeutic route as an adjuvant treatment for all inflammatory dis-ease, and within the current pandemic could help mitigate the risk of cytokine storm associated with severe COVID25.

Reference:

  1. Italiani, P. & Boraschi, D. From Monocytes to M1/M2 Macrophages: Phenotypical vs. Functional Differentiation. Front Immunol 5, 514 (2014).
  2. Davies, L. C., Jenkins, S. J., Allen, J. E. & Taylor, P. R. Tissue-resident macrophages. Nat Immunol 14, 986–995 (2013).
  3. Hu, D. et al. Alleviation of the chronic stress response attributed to the antioxidant and anti-inflammatory effects of electrolyzed hydrogen water. Biochem Bioph Res Co 535, 1–5 (2021).
  4. Gilmore, T. D. The Rel/NF-κB signal transduction pathway: introduction. Oncogene 18, 6842–6844 (1999).
  5. Pires, B. R. B., Silva, R. C. M. C., Ferreira, G. M. & Abdelhay, E. NF-kappaB: Two Sides of the Same Coin. Genes-basel 9, 24 (2018).
  6. Liu, T., Zhang, L., Joo, D. & Sun, S.-C. NF-κB signaling in inflammation. Signal Transduct Target Ther 2, 17023 (2017).
  7. Liu, S. F. & Malik, A. B. NF-κB activation as a pathological mechanism of septic shock and inflammation. Am J Physiol-lung C 290, L622–L645 (2006).
  8. Su, C.-M., Wang, L. & Yoo, D. Activation of NF-κB and induction of proinflammatory cytokine expressions mediated by ORF7a protein of SARS-CoV-2. Sci Rep-uk 11, 13464 (2021).
  9. Courtois, G. & Gilmore, T. D. Mutations in the NF-κB signaling pathway: implications for human disease. Oncogene 25, 6831–6843 (2006).
  10. Albensi, B. C. What Is Nuclear Factor Kappa B (NF-κB) Doing in and to the Mitochondrion? Frontiers Cell Dev Biology 7, 154 (2019).
  11. Oliveira-Marques, V., Marinho, H. S., Cyrne, L. & Antunes, F. Role of Hydrogen Peroxide in NF-κB Activation: From Inducer to Modulator. Antioxid Redox Sign 11, 2223–2243 (2009).
  12. Ye, G. et al. Oxidative stress-mediated mitochondrial dysfunction facilitates mesenchymal stem cell senescence in ankylosing spondylitis. Cell Death Dis 11, 775 (2020).
  13. Yu, H.-C. et al. Down-Regulation of LOC645166 in T Cells of Ankylosing Spondylitis Patients Promotes the NF-κB Signaling via Decreasingly Blocking Recruitment of the IKK Complex to K63-Linked Polyubiquitin Chains. Front Immunol 12, 591706 (2021).
  14. Davies, D. A., Adlimoghaddam, A. & Albensi, B. C. The Effect of COVID-19 on NF-κB and Neurological Manifestations of Disease. Mol Neurobiol 58, 4178–4187 (2021).
  15. Zheng, Z.-G. et al. Hydrogen/oxygen therapy for the treatment of an acute exacerbation of chronic obstructive pulmonary disease: results of a multicenter, randomized, double-blind, parallel-group controlled trial. Respir Res 22, 149 (2021).
  16. Amitani, H. et al. Hydrogen Improves Glycemic Control in Type1 Diabetic Animal Model by Promoting Glucose Uptake into Skeletal Muscle. Plos One 8, e53913 (2013).
  17. Zheng, M. et al. The protective effect of hydrogen-rich water on rats with type 2 diabetes mellitus. Mol Cell Biochem 1–9 (2021) doi:10.1007/s11010-021-04145-x.
  18. Kajiyama, S. et al. Supplementation of hydrogen-rich water improves lipid and glucose metabolism in patients with type 2 diabetes or impaired glucose tolerance. Nutr Res 28, 137–143 (2008).
  19. Ishibashi, T. et al. Therapeutic efficacy of infused molecular hydrogen in saline on rheumatoid arthritis: A randomized, double-blind, placebo-controlled pilot study. Int Immunopharmacol 21, 468–473 (2014).
  20. Ishibashi, T. Molecular Hydrogen: New Antioxidant and Anti-inflammatory Therapy for Rheumatoid Arthritis and Related Diseases. Curr Pharm Design 19, 6375–6381 (2013).
  21. Ohsawa, I., Nishimaki, K., Yamagata, K., Ishikawa, M. & Ohta, S. Consumption of hydrogen water prevents atherosclerosis in apolipoprotein E knockout mice. Biochem Bioph Res Co 377, 1195–1198 (2008).
  22. Qin, S. Role of Hydrogen in Atherosclerotic Disease: From Bench to Bedside. Curr Pharm Design 27, 713–722 (2021).
  23. Shen, N.-Y. et al. Hydrogen-rich water protects against inflammatory bowel disease in mice by inhibiting endoplasmic reticulum stress and promoting heme oxygenase-1 expression. World J Gastroentero 23, 1375–1386 (2017).
  24. Sim, M. et al. Hydrogen-rich water reduces inflammatory responses and prevents apoptosis of peripheral blood cells in healthy adults: a randomized, double-blind, controlled trial. Sci Rep-uk 10, 12130 (2020).
  25. Guan, W.-J. et al. Hydrogen/oxygen mixed gas inhalation improves disease severity and dyspnea in patients with Coronavirus disease 2019 in a recent multicenter, open-label clinical trial. J Thorac Dis 12, 3448–3452 (2020).
  26. Ohsawa, I. et al. Hydrogen acts as a therapeutic antioxidant by selectively reducing cytotoxic oxygen radicals. Nat Med 13, 688–694 (2007).
  27. Chen, M. et al. Hydrogen protects lung from hypoxia/re-oxygenation injury by reducing hydroxyl radical production and inhibiting inflammatory responses. Sci Rep-uk 8, 8004 (2018).
  28. Ning, K., Liu, W.-W., Huang, J.-L., Lu, H.-T. & Sun, X.-J. Effects of hydrogen on polarization of macrophages and microglia in a stroke model. Medical Gas Res 8, 154 (2018).

29. Niu, Y. et al. Hydrogen Attenuates Allergic Inflammation by Reversing Energy Metabolic Pathway Switch. Sci Rep-uk 10, 1962 (2020).

NEXT