Oxidative stress and inflammation in ethiology of acquired metabolic diseases
Oxidative stress is the imbalance between the production and accumulation of reactive oxygen species (ROS) in cells and tissues, and the body’s natural ability to remove excessive free radicals (1). Sustained overproduction of ROS over an extended period of time may lead to damage to the structure and function of cells, also increasing the risk of mutagenesis. Additionally, an excess of reactive oxygen species may initiate the inflammatory process, manifested in the synthesis and secretion of pro-inflammatory cytokines (2).
Closely related: oxidative stress and inflammation, are an important component in the etiology of a number of diseases, such as obesity, insulin resistance, type 2 diabetes, or cardiovascular diseases.
Obesity and its comorbid and interrelated relationships: increased blood pressure, atherogenic dyslipidaemia, and pre-diabetes / diabetes mellitus are key components of the metabolic syndrome (3). The hepatic manifestation of MS is non-alcoholic fatty liver disease (NAFLD). Just like metabolic syndrome, this systemic disorder poses a significant risk of cardiovascular disease (4).
The modification of eating habits plays a huge role in the treatment of these metabolic disorders.
The results of a number of studies indicate that an effective form of prevention and / or therapy of the above-mentioned diseases and their components may be the increase in dietary intake of polyphenols (2).
Antioxidant and anti-innflammatory effects of ARONVIT® – Own research
Products obtained from aronia berry undoubtedly fit into the increasingly popular ‘nutraceutical approach’ (7) to the problem of preventing the aforementioned metabolic disorders.
The aronia berry is distinguished by its high content of polyphenols. As shown by the results of a number of studies, it exceeds the total polyphenol content in other berries, including in raspberry, blackberry, red and blackcurrant (8), blueberry, cranberry and lingonberry (10).
Qualitative (LC-MS) and quantitative (HPLC) analysis of three batches of ARONVIT® extract (9) showed that the polyphenols contained in it include compounds belonging to anthocyanins, flavonols, flavanols, flavan-3-ols and phenolic acids (Figure 1). The dominant group of phenolic compounds identified in our extract are anthocyanins. This is confirmed by the results of published scientific reports (10, 11), which also indicate the quantitative advantage of anthocyanins among phenolic compounds identified in aroniaberry and in its preparations. The result of the qualitative and quantitative identification of anthocyanins in Aronvit® is shown in Figure 2.
Figure 1. Polyphenol structure in ARONVIT®
The determination was carried out by HPLC. The averaged results of analyzes of three separate batches of the extract were presented (Kucharska 2022).
Figure 2. Structure of anthocyanins in ARONVIT®
Based on determinations by HPLC. Averaged results of analyzes of three separate batches of the extract were presented (Kucharska 2022)
The total content of phenolic compounds in 3 series of ARONVIT® extract, determined by the method with the Folin-Ciocalteu reagent, calculated as gallic acid, was 63,458 mg GAE / 100 g DM of the extract (9). This naturally translated into the above-average high antioxidant activity of our extract (Table 1), also confirmed by its ability to significantly reduce lipid peroxidation in mouse RAW 264.7 macrophages (Figure 3).
Figure 3. Effect of chokeberry extract (ARONVIT®) on MDA (malonic aldehyde) levels in LPS-stimulated RAW 264.7 mouse macrophage cells. Statistical significance of P <0.05 compared to the positive control.
- Negative control 2. Positive control 3. Aronvit 0.5 mcg / ml 4. Aronvit 50 mcg / ml 5. Aronvit 500 mcg / ml
In the same model of mice RAW 264.7 cells, cultured for 24h in the presence of various concentrations of ARONVIT® (range 0.5-500 µg / ml), the anti-inflammatory effect of the extract was also demonstrated. Inflammation was induced by the addition of E. coli lipopolysaccharide (LPS).
Figure 4. Effect of ARONVIT® extract on TNF-α and IL-1β levels in LPS-stimulated RAW264.7 macrophage cells. 1. Negative control 2. Positive control 3. Aronvit 0.5 mcg / ml 4. Aronvit 50 mcg / ml 5. Aronvit 500 mcg / ml
The change in the amount of interleukins was significantly different at p <0.05 compared to the positive control. Along with the increase in the concentration of aronia extract, the expression of the tested cytokines, both TNF-α and IL-1β, decreased, with a significant decrease for TNF-α. The conducted studies showed that ARONVIT® aronia extract inhibited pro-inflammatory mediators in a concentration-dependent manner within the optimal dose range. High antioxidant activity and anti-inflammatory effect of polyphenols contained in the aronia extract
are important factors contributing to its pro-health properties (12), including proven beneficial effects in the prevention and / or treatment of metabolic disorders.
Aronia extracts in the prevention and / or therapy of acquired metabolic diseases – state of knowledge
The effectiveness of aronia extracts in the prevention and / or treatment of obesity is well known. In one
of experiments (13) it was shown that phenolic components isolated from A. melanocarpa extract inhibited the process of differentiation of pre-adipocytes of the 3T3-L1 line by inhibiting the expression of PPARγ receptor genes as well as C / EBPα and SREBP1c proteins. As a consequence, when this extract was administered to obese laboratory mice they developed inhibition of obesity and hyperlipidemia, leading to increased glucose tolerance and insulin sensitivity.
Aronia can also be used in the prevention and / or treatment of NAFLD. In mice on a high-fat diet, a significant ability of dry aronia berry extract to inhibit the de novo lipogenesis (↓ TG, ↓ FAS) process in hepatocytes by reducing the mRNA expression of the PPARγ2 receptor gene, lipoprotein lipase (LPL) and aP2 protein. In addition, supplementation with aronia berry extract had a positive effect on the process of restoring redox balance in hepatocytes (↑ SOD activity and ↑ antioxidant potential), also protecting them against damage (↓ ALT and ↓ AST) (5).
The ability of A. melanocarpa extract to lower blood pressure and reduce total cholesterol, confirmed in the latest meta-analysis (14), should be considered extremely important. Many patients are especially hoped for the possibility of supplementing with aronia berry extract as a safe, alternative option in the treatment of dyslipidemia. This is important because the basic therapeutic option, i.e. the use of statins (15), in many patients (especially with hypercholesterolaemia) does not optimize the level of LDL cholesterol. It is also associated with the occurrence of side effects (e.g. muscle pain or life-threatening rhabdomyolysis), prompting the abandonment of long-term pharmacotherapy (16).
The beneficial effects of aronia berries and its preparations on other risk factors for cardiovascular diseases have been described in detail in numerous reviews. Therefore, only the results of selected experimental studies are presented below, perfectly documenting the multidirectional, beneficial effects of polyphenols contained in aronia extracts (17).
The polyphenols contained in aronia extracts are attributed, demonstrated in animal studies, significant effectiveness in increasing the activity of paroxonase-1 (PON1) (18) – an enzyme hydrolysing homocysteine thiolactone, protecting HDL and LDL fractions against oxidation (factor prevention of atherosclerosis), decomposing hydrogen peroxide and oxidized forms of lipids, and initiating the removal of cholesterol from macrophages (19). PON1 activity decrease is mentioned among the risk factors for ischemic heart disease (20, 21).
The following factors predispose to the development of atherosclerosis and coronary artery disease in people with type 2 diabetes: increased concentration of fibrinogen in the blood plasma, related hypercoagulability and chronic inflammation (22). The ability of aronia extract to reduce fibrinogen synthesis was demonstrated in an animal model of induced diabetes. Additionally, the extract showed immunomodulatory effects, manifested in the stimulation of TNF-α and INF-γ secretion by peripheral blood mononuclear cells (PBMC) (23). In another study, methanol aronia extract significantly inhibited glucose absorption in the Caco-2 cell line (↓ 57.1%). On this basis, it was concluded that the observed hypoglycemic effect of the extract may be the result of the inhibition of intestinal transporters of the studied monosaccharide (24).
A number of published studies also indicate the antiplatelet effect of polyphenols present in A. melanocarpa (17). In an in vitro study conducted by Olas (25), the significant ability of A. melanocarpa berry extract to inhibit the activation of platelets, manifested in the inhibition of their adhesion to collagen and reduction of their aggregation, was demonstrated. Additionally, as a strong antioxidant – the extract inhibited the production of the superoxide radical O2- ●, especially in unstimulated platelets. In another experiment, it was additionally shown that aronia extract inhibited the amidolytic activity of two serine proteases: thrombin and plasmin (26).
The impaired normal secretion of nitric oxide (NO) by endothelial cells is also associated with an increased risk of developing cardiovascular diseases. A relatively low concentration of aronia extract significantly induced the synthesis of nitric oxide (NO) and the phosphorylation process of endothelial NO synthase (eNOS) in the bovine coronary artery BCAEC endothelial cell line (27). The ability of aronia extract to induce endothelial-dependent relaxation of blood vessels was also demonstrated in an experiment carried out on isolated porcine coronary arteries. Additionally, at concentrations too low to cause a direct vasorelaxant effect, A. melanocarpa extracts protected the coronary arteries against loss of relaxation following exposure to ROS (28).
- Aleksandrova K i in. Dietary patterns and biomarkers of oxidative stress and inflammation: A systematic review of observational and intervention studies. Redox Biol. 2021 Jun;42:101869. doi: 10.1016/j.redox.2021.101869.
- Hussain T i in. Oxidative Stress and Inflammation: What Polyphenols Can Do for Us? Oxid Med Cell Longev. 2016;2016:7432797. doi: 10.1155/2016/7432797.
- Furman D i in. Chronic inflammation in the etiology of disease across the life span. Nat Med. 2019 Dec;25(12):1822-1832. doi: 10.1038/s41591-019-0675-0.
- Rochlani Y i in. Metabolic syndrome: pathophysiology, management, and modulation by natural compounds. Ther Adv Cardiovasc Dis. 2017 Aug;11(8):215-225. doi: 10.1177/1753944717711379.
- Park CH i in. Aronia melanocarpa Extract Ameliorates Hepatic Lipid Metabolism through PPARγ2 Downregulation. PLoS One. 2017 Jan 12;12(1):e0169685. doi: 10.1371/journal.pone.0169685.
- Byrne CD, Targher G. NAFLD: a multisystem disease. J Hepatol. 2015 Apr;62(1 Suppl):S47-64. doi: 10.1016/j.jhep.2014.12.012.
- Penson PE, Banach M. Natural compounds as anti-atherogenic agents: Clinical evidence for improved cardiovascular outcomes. Atherosclerosis. 2021 Jan;316:58-65. doi: 10.1016/j.atherosclerosis.2020.11.015.
- Benevenuti S. i in. Polyphenols, Anthocyanins, Ascorbic Acid, and Radical Scavenging Activity of Rubus, Ribes, and Aronia. Journal of Food Science 2004, 69: FCT164-FCT169. https://doi.org/10.1111/j.1365-2621.2004.tb13352.x
- Kucharska A. Analiza jakościowa i ilościowa związków polifenolowych w ekstrakcie suchym z owocu aronii Aronvit. Katedra Technologii Owoców, Warzyw i Nutraceutyków Roślinnych Uniwersytetu Przyrodniczego we Wrocławiu. Marzec 2022. Dokument wewnętrzny.
- Zheng W, Wang SY. Oxygen radical absorbing capacity of phenolics in blueberries, cranberries, chokeberries, and lingonberries. J Agric Food Chem. 2003 Jan 15;51(2):502-9. doi: 10.1021/jf020728u.
- Jakobek L. i in. Anthocyanin content and antioxidant activity of various red fruit juices. Deutsche Lebensm-Rundsch. 2007;103(2):58-64
- Banach M i in. Evaluation of Antioxidant and Anti-Inflammatory Activity of Anthocyanin-Rich Water-Soluble Aronia Dry Extracts. Molecules. 2020 Sep 4;25(18):4055. doi: 10.3390/molecules25184055.
- Kim NH i in. Chokeberry Extract and Its Active Polyphenols Suppress Adipogenesis in 3T3-L1 Adipocytes and Modulates Fat Accumulation and Insulin Resistance in Diet-Induced Obese Mice. Nutrients. 2018 Nov 12;10(11):1734. doi: 10.3390/nu10111734.
- Hawkins J i in. Daily supplementation with aronia melanocarpa (chokeberry) reduces blood pressure and cholesterol: a meta analysis of controlled clinical trials. J Diet Suppl. 2021;18(5):517-530. doi: 10.1080/19390211.2020.1800887.
- Grundy SM i in. AHA/ACC/AACVPR/AAPA/ABC/ACPM/ADA/AGS/APhA/ASPC/NLA/PCNA Guideline on the Management of Blood Cholesterol: A Report of the American College of Cardiology/American Heart Association Task Force on Clinical Practice Guidelines. Circulation. 2019 Jun 18;139(25):e1082-e1143. doi: 10.1161/CIR.0000000000000625. Epub 2018 Nov 10. Erratum in: Circulation. 2019 Jun 18;139(25):e1182-e1186.
- Karr S. Epidemiology and management of hyperlipidemia. Am J Manag Care. 2017 Jun;23(9 Suppl):S139-S148.
- Borowska S, Brzóska MM. Chokeberries (Aronia melanocarpa) and Their Products as a Possible Means for the Prevention and Treatment of Noncommunicable Diseases and Unfavorable Health Effects Due to Exposure to Xenobiotics. Compr Rev Food Sci Food Saf. 2016 Nov;15(6):982-1017. doi: 10.1111/1541-4337.12221.
- Lou-Bonafonte JM i in. The Search for Dietary Supplements to Elevate or Activate Circulating Paraoxonases. Int J Mol Sci. 2017 Feb 15;18(2):416. doi: 10.3390/ijms18020416.
- Litvinov D i in. Antioxidant and anti-inflammatory role of paraoxonase 1: implication in arteriosclerosis diseases. N Am J Med Sci. 2012 Nov;4(11):523-32. doi: 10.4103/1947-2714.103310.
- Wang M i in. Quantitative assessment of the influence of paraoxonase 1 activity and coronary heart disease risk. DNA Cell Biol. 2012 Jun;31(6):975-82. doi: 10.1089/dna.2011.1478.
- Zhao Y i in. Association between PON1 activity and coronary heart disease risk: A meta-analysis based on 43 studies. Molecular Genetics and Metabolism 2012;105(1):141-148. https://doi.org/10.1016/j.ymgme.2011.09.018
- Kozek E. i in. Fibrinogen as a coronary risk factor in patients with type 2 diabetes mellitus. Diabetologia Praktyczna 2003;4(4):265–271. Paper in Polish.
- Badescu M i in. Effects of Sambucus nigra and Aronia melanocarpa extracts on immune system disorders within diabetes mellitus. Pharm Biol. 2015 Apr;53(4):533-9. doi: 10.3109/13880209.2014.931441.
- Schreck K, Melzig MF. Traditionally Used Plants in the Treatment of Diabetes Mellitus: Screening for Uptake Inhibition of Glucose and Fructose in the Caco2-Cell Model. Front Pharmacol. 2021 Aug 20;12:692566. doi: 10.3389/fphar.2021.692566.
- Olas B i in. Comparative anti-platelet and antioxidant properties of polyphenol-rich extracts from: berries of Aronia melanocarpa, seeds of grape and bark of Yucca schidigera in vitro. Platelets. 2008 Feb;19(1):70-7. doi: 10.1080/09537100701708506.
- Sikora J i in. Extract of Aronia melanocarpa-modified hemostasis: in vitro studies. Eur J Nutr. 2014 Oct;53(7):1493-502. doi: 10.1007/s00394-014-0653-8.
- Varela CE i in. Effects of a natural extract of Aronia Melanocarpa berry on endothelial cell nitric oxide production. J Food Biochem. 2016 Aug;40(4):404-410. doi: 10.1111/jfbc.12226.
- Bell DR, Gochenaur K. Direct vasoactive and vasoprotective properties of anthocyanin-rich extracts. J Appl Physiol (1985). 2006 Apr;100(4):1164-70. doi: 10.1152/japplphysiol.00626.2005.
- Pavlova V i in. Antioxidant effect of Aronia Melanocarpa extract after doxorubicin treatment. Bulgarian Journal of Agricultural Science 2014;20 (Suppl. 1): 188–192