Nitrosative Stress

This is not medical advice , I am posting this for educational and informational purposes only. This compliments the other post I did on recovering from endotoxemia.

Nitrosative Stress

Is caused by oxidative stress, inflammaton and infection. Those can also lead to Nitrosative Stress which can cause superoxide, and peroxynitrite which are very damaging to the body especially the brain and nervous system. These are a form of Reactive Nitrogen Species. (RNS) Nitric Oxide dependent reactions lead to formation of RNS through the reaction of Nitric Oxide and O2-, peroxynitrite (ONOOH) is formed and leads to oxidation, nitrosation (addition of NO), or nitration (addition of NO2) .

Nitric Oxide is formed from Nitric Oxide Synthase (NOS) which can also produce O2-. Other sources are Xanthine Oxidase, NADPH oxidase, and Myeloperoxidase. There are three types of NOS, inducible nitric oxide synthase (iNOS, NOS2), endothelium NOS (eNOS, NOS3), neuronal NOS (nNOS, NOS1) They no longer use the numerical references to NOS because it inaccurately described them.

Nitric Oxide is a signaling molecule. When functioning properly it is involved with the homeostasis of our blood pressure. Nitric Oxide is released by the endothelial of the smooth muscle to promote smooth muscle relaxation. This increases the diameter of blood vessels to prevent high blood pressure. Oxidative stress and nitrosative stress can deplete NO causing high blood pressure.

Nitric Oxide has antithrombic effects. So if NO is depleted or production is inhibited by free radicals then excessive blood clotting can occur. Impaired NO production can lead to hypertension, preclampsia, diabetes, obesity, erectile dysfunction, and migraine headaches.

NO is generated as a normal part of an immune response. White blood cells known as phocytes have both inducible and inhibitory enzymes to control NO. NO secreted as part of the immune response uses NOS free radical properties and it is toxic to bacteria, causing DNA damage and degredation of the iron sulfur center.

NO increases expression of superoxide dismutase which catalyzes superoxide anion to hydrogen peroxide. NO upregulates Heme oxygenase 1 and ferritin expression which reduce anion concentrations in blood vessels. This prevents blood vessels from leaking. Inflammation , endotoxins, farm chemicals and many other things can inhibit this. This leads to hardening of the arteries because the body will use cholesterol and calcium to plug the leaks.

NO helps convert thiol groups including cysteine residues in proteins, to form s-nitrosothiols. Thiols are organo sulfur compounds. Our bodies need sulfur for many things and all cell need it. If sulfite levels get high from BH4 , CBS issues, gut dysbiosis or candida it could interfere with the proper function of the thiol groups. S-nitrosylation is an important mechanism for dynamic, post translational regulation of most all major classes of proteins. Post translational regulation is control of gene expression.

Guanylyl known as the NO receptor. Activate guanylyl cyclase enables the production of the cyclic guanosine monophosphate (cGMP) . cGMP activates and regulates the enzyme G-kinase. G-kinase modifies proteins in cell through phosphorylation of the myosin light chain kinase which causes smooth muscle relaxation.

Chronic expression of NO can lead to cancer of the epithelium, juvenile diabetes, multiple sclerosis, arthritis, and ulcerative colitis. NO is an oxidising free radical and does cause some oxidative stress on the bodies membranes and tissues. Sustained No production can result in tissue toxicity but most damage is caused from NO causing peroxynitrite formation.

NO protects organs from ischemia, but can contribute to reperfusion injury by increasing inflammation and oxidative damage.

NO forms complexes with transitional metal ions, including those regularly found in metalloproteins. The main trap for no is oxyhemoglobin (oxygen loaded form of hemoglobin) , which binds to NO faster then oxygen. The reaction produces methaemoglobin other NO sensitive metalloproteins are NOS, Cytochrome P450, Ferritin, Ceruloplasmin, mitochondrial respiratory chain. NOS is discussed later, Cytochrome P450 is a very important for detoxing the body. Ferritin helps regulate iron homeostasis. Ceruloplasmin is a copper-carrying protein that is involved in iron metabolism. Mitochondrial respiratory chain is involved in cellular energy.

As NO rises it oxidizes hemoglobin. In healthy individuals about 1% of total hemoglobin has been oxidized mainly by NO. Excess oxidative stress causes excess NO. which can make methemoglobin higher. Methemoglobin contains Fe3 which cannot bind to oxygen. Curcumine and Lactoferrin reduces Fe3 levels in the body. Curcumin can make you anemic if taken for too long because it can also chelate the good iron from the body. Excess methemoglobin is called acquired methemoglobinemia. This can cause skin discoloration, shortness of breath, lethargy, headache, dizzyness, brain fog. Methylene blue helps with this because it is caused by sepsis but those who are G6PD deficient cannot take methylene blue it could kill them.

Some causes of high NO levels.

Prolonged or frequent immune system activation. Which is common in those with CFS, MS, and Fibromyalgia. The most common cause is viral infection. But any number of infections can cause this especially mycoplasma. So to reduce peroxynitrite levels it is important to reduce the viral load on the body, calm the immune system and reduce inflammation.

Inflammation and pro-inflammatory cytokines.

Effects of low NO levels

Low levels of NO will cause low levels of ATP which means the cells would have less energy. This also disables autophagy preventing recycling of mitochondria.

iNOS

NOS is calcium dependent. iNOS is primarily expressed in macrophages a type of white blood cell of the immune system that engulfs and digest cellular debris, foreign substances, microbes, cancer cells and any other cell that does not the type of proteins specific to healthy cells. They are found in almost all tissues in the body. They are expressed primarily after induction by cytokines(small proteins involved in cell signaling) , especially IFN-Y and lippopolysaccharide (endotoxins) . Lippopolysaccharide is abbreviated LPS. Because iNOS contributes to inflammation and destroys parasites and infections it is a major contributor in septic shock. Excessive iNOS can lead to vascular permeability.

Reactive oxygen species works together with reactive nitrogen species to damage cells. Reactive oxygen species produces hydrogen peroxide which can be very damaging to the body. Reactive nitrogen produces peroxynitrite which is also very damaging to the body. Normally the body can neutralize them but when under excess oxidative stress from emotional stress, physical stress, infection, toxins or from nutritional deficiencies the body cannot neutralize and detox the reactive nitrogen and reactive oxygen species. It is a cycle that can be difficult to break. The inflammation that is caused prevents the body from being able to detox and also the metabolic issues it causes puts an even heavier burden on the body because it produces toxins adding to the burden. This is why it is very important to address the nutrient deficiencies caused by illness or toxins and to reduce inflammation.

eNOS

Endothelial NOS (eNOS) is primarily responsible for the generation of NO in vascular endothelium. Endothelium is a monolayer of cells lining the interior surface of blood vessels. No produced by eNOS in the vascular endothelium plays a crucial role in regulating vascular tone, cellular proliferation (process that results in an increase in the number of cells), leukocyte adhesion (white blood cell), and platelet aggregation. eNOS is dependent on calcium and synthesizes NO from L-arginine and oxygen which binds to the heme group of eNOS. Heme is the part of red blood cells that contain the iron which is needed to transport oxygen. The binding of Tetrohydrobiopteran (BH4) is essential for eNOS to generate NO. In the abscense of BH4 which many toxins like glyphosate, mercury and endotoxins can cause eNOS shifts from a dimeric to a monomeric form, thus becoming uncoupled. This causes a production of superoxide anion by eNOS instead of NO. It is a highly reactive free radical that damages the cardiovascular system. This also causes phenylketonuria, ammonia build up in the body because phenylalanine can no longer be recycled. Eating things high in phenylalanine like meat or potatoes will increase the ammonia levels making a person suffering from this sicker, their urine will get dark and smell and when very bad their body odor will be strong.

eNOS causes a reduction in Cationic Amino Acid Transporter 2 (Cat2) which mediates the L-arginine transport needed for raised NO production.

When functioning properly eNOS prevents hardening of the arteries and damage of the blood vessles. It has anti-artheriosclerotic effects. Oxidative stress can cause it to become uncoupled causing many health problems because this also increase reactive oxygen species. This is the one that will cause issues throughout the body because many metabolic pathways will be effected. There are natural ACE inhibitors that will reduce the damage when this happens. Remember the goal is homeostasis. Once the issues that have caused it’s uncoupling have been addressed focus on healing. To continue to take allopathic or natural things to treat symptoms keeps you on a hamster wheel because you have to address the cause. Treating the symptoms will not resolve the problem it will only reduce the symptoms. Enos is very important to the healing of soft tissue throughout the body so it is very important for all pathways to be functioning. Through indirect processes fluoroquinolones also inhibit BH4 and interfere with eNOs. BH4 is involved in the production of serotonin, and L-Tyrosine so inhibition or uncoupling can have dramatic effects on our brains.

Neuronal NOS

Neuronal NOS which used to be known as NOS1 is constitutively(always active) expressed in the central(brain and spinal cord) and peripheral (nerves throughout the rest of the body) neurons and some other cell types.

nNOS functions include synaptic plasticity in the Central Nervous System, central regulation of blood, and vasodilation via nitrergic nerves.

Peroxynitrite

Peroxynitrite is formed when NO reacts with superoxide (O2-) which it readily combines with. The higher the levels of NO present the great the risk of it reacting with super oxide to form peroxynitrite. Peroxynitrite can cause oxidation to hemoglobin causing anemia.

When peroxynitrite is formed it depletes the bioactivity of NO which effect the bodies ability to regulate blood pressure, platelet activation, and vascular cell signaling. This can result in high blood pressure, and hardening of the arteries. When NOS enzymes have limited L-arganine or BH4 they my produce superoxide instead of NO. The can be transformed to peroxynitrite. BH4 reacts with peroxynitrite and is oxidized to biopterin. BH4 is also oxidized by neopterin which is an indication of an immune response. This causes inflammation which in turn can stimulate more peroxynitrite synthesis.

Peroxynitrite can react with other components of the cell including lipids(fats), thiols(organosulfur compounds) , amino acid residues(an amino acid contained within a peptide or protein) , DNA bases (adenine, cytosine, guanine,thymine) , and antioxidants.

Peroxynitrite can react with other molecules to form additional types of RNS including nitrogen dioxide (N2O2) as will as other chemically reactive free radicals which include peroxynitrous acid, hydroxyl radical, carbon dioxide, carbanate radical and dinitrogentrioxide.

Peroxynitrite can react directly with proteins that contain transition metal centers. Therefore it can modify proteins such as hemoglobin, myoglobin, and cytochrom C by oxidising heme into it’s corresponding ferrous forms. Two of the most common reactions is cystein oxidation and tyrosine nitration. These reactions affect protein structure and function so have the potential to cause changes in the catalytic activity of enzymes, altered skeletal organization and impaired cell signal transduction. So it can hinder the bodies ability to heal and examples of illnesses caused by changes in the catalytic activities of enzymes are illnesses such as porphyria and phenylketonuria. Also with the cells ability to communicate impaired all heck can break loose.

Both NO and peroxynitrite can oxidize the iron-sulfur protein based enzymes in the Krebs cycle (citric acid cycle). The iron-sulfur protein is known as Aconitase. Krebs cycle is involved in producing cellular energy. It is the metabolic pathway that connects carbohydrate, fat and protein metabolism. So problems with this pathway would have dramatic effects throughout the body. So we can see the importance of reducing peroxynitrite levels.

No reacts with nitrogen dioxide (NO2) presumably instead of superoxide to form Dinitrogen Trioxide (NO3) , which is a type of reactive nitrogen species. All nitrogen oxides are good oxidizing agents. This cause excess Cyclic GMP, vascular collapse and septic shock, s-nitrothiol formation on proteins (thionitrites).

Now you should be able to see why iNOS has been implicated in inflammatory bowel disorders.

GTP cyclohydrolase increases BH4, folate is needed to produce it. Do not take folic acid it is man made and can build up in the body causing health problems.

Vitamin C the real vitamin C not man made, Vitamin E and Folate not to be mistaken for Folic Acid which is man made and can build up in the body damaging the NMDA receptor and increasing the risk for cancer. These all scavenge peroxynitrite.

Hydroxycolbamin scavenges peroxynitrite

COX2 inhibitors reduce peroxynitrite. In my list of supplements I listed some things that will inhibit COX.

Getting rid of infection and reducing inflammation reduces peroxynitrite.

Reducing toxins in the body especially glyphosate and heavy metals reduces peroxynitrite.

Ellagic Acid reduces peroxynitrite.

https://www.sciencedirect.com/science/article/pii/S0308814608006195

How Nitric Oxide Species are produced.

https://www.sciencedirect.com/science/article/pii/S0005272899000286

Disregulation of NO can cause many health problems from diabetes to cancer.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3345541/

Though I do not agree with statin use because it does more damage to the body then oxidative stress does, this article has a good summary of Nitric Oxide production and the enzymes that regulate it.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3345541/

iNos has been implicated in inflammatory bowel disease.

https://gut.bmj.com/content/45/2/199

Inflammation and infection are known causes of hardening of the arteries. The blood vessels start to leak and the body uses calcium and cholesterol to seal the leaks resulting in hardening of the artieries. INOS causes leakage in blood vessels in response to inflammation and infection. This is why it is very important to address the infection and inflammation.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1572034/

Excess iNOS production can cause anemia.

http://jultika.oulu.fi/Record/isbn951-42-6851-2

eNOS issues have been linked to cardiovascular disease, erectile dysfunction, hypertension, diabetes, and obesity.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3345541/

This is cause by oxidative stress .

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3174401/

Toxins and infection can lead to excess NMDA receptor activation which can lead to nitrosative stress.

https://www.researchgate.net/profile/Zezong_Gu2/publication/42441952_Redox_Reactions_Induced_by_Nitrosative_Stress_Mediate_Protein_Misfolding_and_Mitochondrial_Dysfunction_in_Neurodegenerative_Diseases/links/575a2c2e08ae9a9c954f2c86/Redox-Reactions-Induced-by-Nitrosative-Stress-Mediate-Protein-Misfolding-and-Mitochondrial-Dysfunction-in-Neurodegenerative-Diseases.pdf?origin=publication_detail

Epselen can scavenge peroxynititrite but has not been approved for usage for anything but mental disorders.

https://www.sciencedirect.com/science/article/pii/S1054358908609862

Thymoquinone reduces oxidative stress which damages the brain.

https://www.ncbi.nlm.nih.gov/pubmed/28655600

https://www.ncbi.nlm.nih.gov/pubmed/24814387

Baicalein found in sculletaria and plantain herb protects from peroxynitrite damage.

https://www.ncbi.nlm.nih.gov/pubmed/27518088