Pomegranate peel catechins – inhibition of NET formation

We could show that the flavonoids (−)-epicatechin, (+)-catechin hydrate, and rutin trihydrate as well as vitamin C and the pharmacological substances N-acetyl-L-cysteine and 5-aminosalicylic acid inhibited PMA induced ROS production and NET formation.” (1)

Why care about NET formation? because it may be associated with worse risk of coagulation and blood clots forming in severe COVID-19 illness. Interleuken-6 (IL-6) can increase NET formation by neutraphil white blood cells attempting to ensnare pathogens and IL-6 levels tend to be elevated in mor severe COVID-19 illness.

NET-neutraphil extracellular traps: “Thrombosis [coagulation/blood clotting], sepsis, and thick mucous secretions share neutrophil extracellular traps as a common causal agent. In addition to phagocytosis and degranulation, neutrophils can kill pathogens by extruding neutrophil extracellular traps (NETs), a process termed NETosis.” (2)

Reference List

  1. Kirchner T, Hermann E, Möller S, et al. Flavonoids and 5-aminosalicylic acid inhibit the formation of neutrophil extracellular traps. Mediators Inflamm. 2013;2013:710239. doi:10.1155/2013/710239 https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3871909/
  2. Yanuck SF, Pizzorno J, Messier H, Fitzgerald KN, Evidence Supporting a Phased Immuno-physiological Approach to COVID-19 From Prevention Through Recovery. athmjournal.com/ https://athmjournal.com/covid19/research/evidence-supporting-a-phased-immuno-physiological-approach-to-covid-19-from-prevention-through-recovery/ *”interventions for one phase may not be appropriate for a different phase. The 4 phases addressed are: Prevention, Infection, Inflammation & Recovery.

Pomegranate, neuroinflammation, antimicrobial, metal chelator.

Pomegranate and its various products all have been found to have beneficial phytonutrients. The antioxidant power of the peel is strong enough to protect metal from rusting. The juice also includes polyphenol antioxidants that can enter the blood brain barrier (1, 4) and help detoxify oxidative stress chemicals within the brain (1 4), – anti-corrosive for the brain.

A metabolite (UA) of the pomegranate polyphenols was measured in brain tissue and blood plasma during an animal study. (4) “The concentration of UA in the brain was 1.68 ± 0.25 ng/g tissue and in plasma 18.75 ± 3.21 ng/mL. ” (4) It also seemed to increase activity of an anti-inflammatory enzyme (“GPx activity“) which would increase our own antioxidant production and helped reduce levels of the amyloid-β protein that is thought to be involved in Alzheimer’s dementia . (4)

  • It seems that PJ [pomegranate juice] caused down-regulation of α-synuclein protein expression since in rats treated alone with the juice, the levels of α-synuclein oligomers was decreased while the ratio of this fraction to monomeric was higher than in Control. This effect is likely to be involved in the improvement of neuronal cell survival, which was also reported for other natural preparations [42,62]. ” (4)

Pomegranate extract helped reduce/improve the balance of levels of the amyloid-β proteins involved in Alzheimer’s dementia in an animal model of the disease. (2) “The mice (age 24-30 mo) received either PE [pomegranate extract] (at 100 & 200 mg/kg) or a control solution daily for three weeks” (2)

What is standardized pomegranate extract? And how might you calculate a human dose?

It is hard to know and dose might vary with the type of pomegranate extract. Whether the seed, juice or peel, or the whole fruit is used to make the pomegranate extract might vary – read each article for details. Pomegranate Peel Extract (PPE) is used regularly in research regarding the medical uses.

  • Doses of 100 or 200 mg/kg of “a standardized pomegranate extract (PE)” was used in a study of an aged Alzheimer’s disease animal model. Ratios of the amyloid-beta proteins thought to be involved in Alzheimer’s disease were improved. (2) “Our findings support a specific anti-amyloidogenic mechanism of a pomegranate extract in this aged AD animal model. ” (2)
  • Pomegranate seeds (PGSE) were used to make the extract for an animal study of ischemic stroke damage to the brain. Doses of 100 or 200 mg/kg were found protective against oxidative damage. (1) An upper tolerance dose was trialed at 800 mg/kg but it did not cause toxic effects.
    • No toxicity was observed even with high-dose PGSE consumption (800 mg/kg, for 14 days). ” (1)

Without knowing how the extract was made, roughly an adult size amount (for a person weighing 70 kilograms/154 pounds) might be 7-14 grams per day (approximately 2-4 teaspoons).

I found 1-2 tablespoons (3-6 teaspoons) of my whole peel extract to be a beneficial amount for my symptoms. It can have diuretic side effects, use early in the day and have some extra nondiuretic fluids. Using only the inner white part of the peel can reduce the diuretic effects without losing other health benefits – the ratio of phytonutrients is slightly different for the inner and outer part of the peel, with both containing some of the hyrolyzable tannins that have healing, and diuretic effects. For directions/recipe and more information see: G13. Pomegranate Benefits and Preparation.

Pomegranate juice also was found neuroprotective in an Alzheimer’s animal model study. “Commercial 6-fold concentrated pomegranate juice (PJ) […] The total polyphenols content expressed as g of ellagic acid (EA) equivalents per L of juice was 18.90 ± 0.96 g/L. (4)

For humans eight ounces of pomegranate juice daily for four weeks was found beneficial for memory and changes in blood tests suggested it was being absorbed in an amount sufficient to help against inflammation.

In older subjects with age-associated memory complaints, who drank 8 ounces of pomegranate juice for four weeks, a significant improvement in verbal and visual memory as well as an increase in plasma Trolox-equivalent antioxidant capacity was observed. Noteworthily, individuals drinking pomegranate juice represented an increased level of a metabolite of pomegranate ellagitannins—urolithin A glucuronide—in plasma [15]. It is believed that pomegranate’s neuroprotective effects are mediated by urolithins—the colonic microbiota ellagitannins (ETs)-derived metabolites [8].” (4)

From my own use I have found about four ounces per day beneficial for mood and four ounces twice a day was helpful on a bad day. The fresh seeds, 1/3-1/2 cup also helped my anxiety symptoms in a similar way. Dried pomegranate seeds, anardana, are available in markets with foods from India, about 2-3 tablespoons might be an equivalent serving.

Pomegranate extracts may also have healing benefits when used topically, for wound care in this study:

  • Whole fruit extract was used topically, diluted to 7.5% was mentioned in an animal based study on wound healing. “…whole fruit extract of pomegranate (P. granatum Linn.) standardized with 40% ellagic acid produced by Xi’an Bio-Technology Co., Ltd.,…” (3)

Regarding CoV19 & pomegranate peel

In the case of SARS-CoV-2/COVID-19 there may be a problem with the virus suppressing the normal immune response of interleuken I and III (ref) Inflammatory interleuken 1 and 6, IL-1 and IL-6, can become elevated in patients with severe illness. Pomegranate peel or whole fruit extract has been found to have significant anti-inflammatory and immune-modulating effects. In different studies it has been shown to have benefits by inhibiting levels of IL-1, (7) , and IL-6. (5)

  • PGPE consumption […] prevent the instigation of a low-grade inflammatory status compared to OVX Control animals, through the down-regulated expression of CCL2 (0.33-fold; p < 0.05) & IL1-R1 (0.64-fold; p < 0.001) and the up-regulated expression of IL1-Rn (1.89-fold; p < 0.001)” (7)
    • IL1-Rn= “IL-1 receptor antagonist allows unopposed IL-1 activation” (6)
  • In another study, we demonstrated that PFE significantly inhibited the excessive production of IL-6 and IL-8 via suppression of the JNK-, extracellular signal-regulated kinases (ERK)- MAPKs and NF-κB-signaling events. (19)  ” (5)

*preliminary version.

Disclaimer: This information is provided for educational purposes within the guidelines of Fair Use. It is not intended to provide individual guidance. Please seek a health care provider for individualized health care guidance.

Reference List

  1. Sarkaki A, Rezaiei M, Gharib Naseri M, Rafieirad M. Improving active and passive avoidance memories deficits due to permanent cerebral ischemia by pomegranate seed extract in female rats. Malays J Med Sci. 2013;20(2):25–34. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3743996/
  2. Ahmed AH, Subaiea GM, Eid A, et al., Pomegranate extract modulates processing of amyloid-β precursor protein in an aged Alzheimer’s disease animal model. Curr Alzheimer Res. 2014;11(9):834-43. https://ncbi.nlm.nih.gov/pubmed/25274111
  3. Yuniarti WM, Primarizky H, Lukiswanto BS. The activity of pomegranate extract standardized 40% ellagic acid during the healing process of incision wounds in albino rats (Rattus norvegicus). Vet World. 2018;11(3):321–326. doi:10.14202/vetworld.2018.321-326 https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5891847/
  4. Kujawska M, Jourdes M, Kurpik M, et al. Neuroprotective Effects of Pomegranate Juice against Parkinson’s Disease and Presence of Ellagitannins-Derived Metabolite-Urolithin A-In the Brain. Int J Mol Sci. 2019;21(1):202. Published 2019 Dec 27. doi:10.3390/ijms21010202 https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6981883/Commercial 6-fold concentrated pomegranate juice (PJ) was obtained from Alter Medica (Żywiec, Poland). The product was manufactured in accordance with the principle of HACCP (hazard analysis and critical control point) and fruit ingredients are fully compliant with the Code of Practice of the European Fruit juice Association (AIJN). Since pomegranate’s ellagitannins and their hydrolysis product—ellagic acid—have been demonstrated to be precursors of potentially neuroprotective urolithins, including urolithin A [8], which we detected in this study, we identified these phenolics in the tested juice. The ellagitannin composition of PJ was as follows: Galloyl-hexoside, ellagic acid-hexoside, 3-bis-HHDP-hexoside (pedunculagin), 4-galloyl-bis-HHDP-hexoside (casuarinin), and ellagic acid (Figure S1). The total polyphenols content expressed as g of ellagic acid (EA) equivalents per L of juice was 18.90 ± 0.96 g/L. ” *so full strength juice might have a sixth or approximately 3 grams/L of ellagic acid.
  5. Rasheed Z. Intake of Pomegranate Prevents the Onset of Osteoarthritis: Molecular Evidences. Int J Health Sci (Qassim). 2016;10(2):V‐VIII. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4825888/In another study, we demonstrated that PFE significantly inhibited the excessive production of IL-6 and IL-8 via suppression of the JNK-, extracellular signal-regulated kinases (ERK)- MAPKs and NF-κB-signaling events. (19) “
  6. IL1RN interleukin 1 receptor antagonist [ Homo sapiens (human) ], Gene ID: 3557, updated on 17-Apr-2020 https://ncbi.nlm.nih.gov/gene/3557
  7. Mélanie Spilmont, Laurent Léotoing, Marie-Jeanne Davicco, et al., Pomegranate Peel Extract Prevents Bone Loss in a Preclinical Model of Osteoporosis and Stimulates Osteoblastic Differentiation in Vitro, Nutrients 2015, 7(11), 9265-9284; https://doi.org/10.3390/nu7115465 https://www.mdpi.com/2072-6643/7/11/5465/htm
  8. **helped “encephalitis lethargica symptoms during ME(ME/CFS) ” anecdotal.

Today, it is well-known that the TRPA1, TRPC5, TRPM2, TRPM7, TRPV1, TRPV3, and TRPV4 cation channels are activated by ROS and reactive nitrogen species (RNS) [2932].” (Nazıroğlu 2015)

Anemia of Inflammation, IL-6, Hepcidin, Iron and Vitamin C.

During bacterial infections iron is shifted out of hemoglobin production and into ferritin and intracellular storage. (Iron and Anemia of Chronic Inflammation, post) If amounts of the free iron overloads the cell capacity and antioxidant capacity then cell death may result and other damage. The shift of iron involves an increase in the inflammatory cytokine IL-6 (Interleucon-6) which causes an increase in hepcidin, which causes the removal of iron from hemoglobin. (5)

Having extra antioxidants in the diet can help the body cope with the sudden increase in free iron – think of rust – iron oxidizes and the body doesn’t want rust anymore than you want it on your car. Antioxidants like vitamin A and C and bioflavonoids can all help stabilize the chemically active iron. The anti-malarial herbal extract called artemisinin may help by binding some of the excess free iron stores. (4)

ACE2 receptors are likely more prevalent in the lung cells of smokers (1) and COPD patients, (3), and people living in air pollution because it has a protective effect against lung injury. It helps decrease levels of the inflammatory cytokine IL-6. (2) Which suggests that extra vitamin C and bioflavonoids, and other nutrients may also help reduce the risk of anemia of chronic inflammation and other lung and heart risks from chronic hypoxia (low oxygen levels in the blood).

The COVID-19 illness may have a risk of sudden death due to cardiovascular problems upon relapse/reinfection (6) because of the shift in iron and resulting risk of cell death due to excess free iron. The use of iron chelators or artemisinin and continuing the use of extra antioxidants such as vitamin C and bioflavonoids might help prevent excess coagulation and reverse the shift of iron atoms from hemoglobin molecules to ferritin and free iron within cells. Infectious microbes need iron too so it is a self defense strategy that may become excessive in the case of severe COVID-19 illness.

There is more about bioflavonoids, vitamin C, and coagulation risks of SARS-CoV-2 infection in this post: Bitter Taste Receptors in the Lungs – Hesperidin’s Decongestant Properties.

Reference List

  1. SJ Brake, K Barnsley, Wenying Lu, et al., Smoking Upregulates Angiotensin-Converting Enzyme-2 Receptor: A Potential Adhesion Site for Novel Coronavirus SARS-CoV-2 (Covid-19). J. Clin. Med.20209(3), 841; https://doi.org/10.3390/jcm9030841 https://www.mdpi.com/2077-0383/9/3/841/htm
  2. Lin CI, Tsai CH, Sun YL, Hsieh WY, Lin YC, Chen CY, Lin CS. Instillation of particulate matter 2.5 induced acute lung injury and attenuated the injury recovery in ACE2 knockout mice. Int J Biol Sci 2018; 14(3):253-265. doi:10.7150/ijbs.23489. Available from http://www.ijbs.com/v14p0253.htm air pollution studies – ACE2 would be protective against ACE->IL-6 “Increased ACE levels cause the activation of AT1R by Ang II, consequently inducing the expression of the inflammatory cytokines IL-6, TNF-α and TGF-β1 [17]. ACE2 counter-regulates the effects produced by Ang II by converting Ang II to Ang-(1-7), activates Mas to repress the signaling pathways of STAT3 and extracellular signal-regulated kinases (ERK) [18], and acts as an anti-inflammatory factor [19].
  3. ACE-2 Expression in the Small Airway Epithelia of Smokers and COPD Patients: Implications for COVID-19. https://erj.ersjournals.com/content/early/2020/03/26/13993003.00688-2020
  4. Inhibitory effect of novel iron chelator, 1-(N-acetyl-6-aminohexyl)-3-hydroxy-2-methylpyridin-4-one (CM1) and green tea extract on growth of Plasmodium falciparum https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4590262/Artemisinin found in the Chinese medicinal plant (Artemisia annua) binds iron to form ferric-dihydroartemisinin complex, resulting in reactive oxygen species (ROS)-mediated potent anti-malarial activity against ring and late stage of CQ-resistant P. falciparum malaria parasites [5]. “
  5. Covid-19 – Liver – iron overload – lung cascade – coagulation a forum topic on peakprosperity.com with links to research, excerpts and discussion re COVID-19 – Liver – iron overload – lung cascade – coagulation. https://www.peakprosperity.com/forum-topic/covid-19-liver-iron-overload-lung-cascade-coagulation/
  6. Jules Quartly, Exclusive: Chinese doctors say Wuhan coronavirus reinfection even deadlier. Feb 14, 2020, TaiwanNews.com, https://www.taiwannews.com.tw/en/news/3876197