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Edible Fruits Extracts Affect Intestinal Microbiota Isolated from Patients with Noncommunicable Diseases Associated with Chronic Inflammation

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Edible Fruits Extracts Affect Intestinal Microbiota Isolated from Patients with Noncommunicable Diseases Associated with Chronic Inflammation Experimental articles UDC 579.61 https://doi.org/10.15407.biotech13.05.087 EDIBLE FRUITS EXTRACTS AFFECT INTESTINAL MICROBIOTA ISOLATED FROM PATIENTS WITH NONCOMMUNICABLE DISEASES ASSOCIATED WITH CHRONIC INFLAMMATION T. V. Meleshko1, 2 1 Uzhhorod National University, Department O. V. Pallah1, 2 of Clinical Laboratory Diagnostics and Pharmacology, R. O. Rukavchuk2 Faculty of Dentistry, Ukraine L. S. Yusko1, 2 2 Uzhhorod National University, N. V. Boyko1, 2 Research Development and Educational Centre of Molecular Microbiology and Mucosal Immunology, Ukraine E-mail: [email protected] Received 27.07.2020 Revised 08.10.2020 Accepted 31.10.2020 The aim of our study was to investigate the gut microbiota in patients with noncommunicable di seases associated with chronic inflammation, including obesity, type 2 diabetes, atherosclerosis, and cardiovas- cular disease as well as to find out potential ability of edible plants fruits extracts to inhibit the growth of selected conditionally pathogenic microorganisms. Limited clinical trial was performed and gut microbiota analysis was done using routine methods and qPCR. The antibacterial properties of edible plants fruits in relation to the selected potentially patho- genic microorganisms were studied. The composition of the intestinal microbiota of obese patients was characterized by an increase in the number of Enterococcus spp. and Lactobacillus spp. along with a decrease in the amount of Escherichia coli. Decreases in E. coli and lactobacilli were observed in patients with type 2 diabetes. In atherosclerosis, an increase in streptococci, enterococci, and enterobacteria was observed, whereas in patients with cardiovascu- lar disease there was an additional increase in staphylococci and candida along with a decrease in E. coli. Decreases in Bifidobacterium spp., Bacteroides spp., Roseburia intestinalis and Akkermansia muciniphila were observed in patients of all groups. The growth of Klebsiella spp. was inhibited by red currant (Ribes rubrum) and plum (Prunus domestica) extracts; Enterobacter spp. — cherry (Prunus avium) extract; Proteus spp. — extracts of blueberry (Vaccinium myrtillus) and dogwood (Cornus mas); Staphylococcus spp. — the extracts of black currant (Ribes nigrum), cherry (Prunus avium), plum (Prunus domestica), jostaberry (Ribes nigrum Ribes divaricatum Ribes uva-crispa), cherry plum (Prunus cerasifera) and dogwood (Cornus mas). The obtained data can be used for early diagnosis of noncommunicable diseases and for their preven- tion with the help of personalized nutrition. Key words: ob esity, type 2 diabetes mellitus, atherosclerosis, cardiovascular diseases, gut micro- biota, edible plants fruits. According to the World Health deaths in the world [1]. NCDs result from a Organization (WHO), non-communicable combined influence of genetic, physiological, diseases (NCDs) are chronic diseases that environmental, and behavioral factors [2]. are not transmitted from person to person, Studies of changes in the intestinal have a long course, and progress slowly. In microbiome and its role in the occurrence the late twentieth century, NCDs turned of NCDs have become extremely relevant in into a global epidemic and one of the greatest recent years [3]. Microbiome is part of human threats to human life and health. According physiology and is significantly involved in a to the WHO, 40 million people die annually wide range of vital physiological processes, from NCDs, which accounts for 70% of all including energy homeostasis and metabolism, 87 BIOTECHNOLOGIA ACTA, V. 13, No 5, 2020 synthesis of vitamins and other important fruits which are characterized by high nutrients, endocrine signaling, prevention biologically active compounds (BAC) of colonization by pathogens, regulation contents and ability to stimulate the growth of immune function, and metabolism of of beneficial microorganisms and inhibit xenobiotics, carcinogens, and other harmful the growth of conditionally pathogenic compounds [4]. microorganisms could be perspective Persistent low-grade inflammatory components for personalized nutrition. response underscores metabolic syndrome and Therefore, the aim of our research was to is a risk factor for cardiovascular diseases study intestinal microbiota in patients with (CVDs) [5, 6]. Inflammatory markers are NCDs related to chronic inflammation, namely associated with obesity and the risk of obesity- obesity, T2D mellitus, atherosclerosis, and related CVDs [7]. Perturbation of intestinal CVDs as well as to find out potential ability microbiota and changes in gut permeability of edible plants fruits extracts to inhibit the are triggers for the chronic inflammatory growth of selected conditionally pathogenic state [8]. “Metabolic endotoxaemia” is a term microorganisms. used to describe a link between gut bacteria, endotoxins, and their circulating levels, with Materials and Methods inflammatory-induced obesity and metabolic diseases linking it to CVDs [9]. Participants and study design Some research studies [10] demonstrated In order to study gut microbiota in patients that intestinal microbiota changes related to with NCDs related to chronic inflammation, obesity lead to threshold inflammation. In we performed a limited clinical case study, in obese people, intestinal microbiota changes which four groups were formed: 1 — patients stimulate the absorption of monosaccharides with obesity; 2 — patients with type 2 diabetes; due to the increased number of capillaries 3 — patients with atherosclerosis; 4 — patients in the small intestine epithelium [11] and with cardiovascular diseases. In order to significantly increase the ability to obtain achieve this goal, we examined 10 people from more energy from food by increasing the each group. number of microorganisms capable of The inclusive criteria for obesity were the fermenting indigestible carbohydrates in the value of the body mass index (BMI), which colon [12, 13]. Obesity is a major risk factor exceeds () 30 kg/m2 [19]; signed informed for the development of type 2 diabetes (T2D), consent to participate in the study. Exclusion which leads to the destruction of insulin criteria: smoking, alcohol or drug use, diabetes receptors and causes resistance to insulin. In mellitus, CVDs, clinically significant kidney turn, patients with diabetes also tend to suffer or liver disease (or other organs and organ from comorbidities, such as hypertension and systems), acute inflammatory diseases at dyslipidemia, which further accelerates the the time of examination, cancer; significant atherosclerotic process, and, therefore, such lifestyle changes, mainly of dietary habits and patients have an extremely high cardiovascular physical activity in the period shorter than 6 risk [14]. Atherosclerosis is a major risk months. factor for CVDs assuming accumulation of Patients with T2D were selected according cholesterol and macrophages on arteries to the criteria typical of this nosology [20]: walls, thus contributing to the formation fasting plasma glucose 6.1 mmol/l; impaired of atherosclerotic plaques [15]. Recent glucose tolerance — two hours after the oral studies suggest that intestinal microbiota dose a plasma glucose 7.8–11.1 mmol/l; disruption may also enhance development of glycated hemoglobin (HbA1c) 6.5%; signed atherosclerosis and CVDs [16, 17]. informed consent to participate in the study. In our opinion, among numerous NCDs, it Exclusionary criteria were smoking, alcohol, is necessary to distinguish a group of diseases or drug abuse; pregnancy; an unstable medical directly relating to changes in the microbiome status; significant lifestyle changes, mainly and the main trigger of which is chronic of dietary habits and physical activity in the inflammation, that is obesity, T2D mellitus, period shorter than 6 months. No participants atherosclerosis, and CVDs. had clinically significant cardiovascular, renal Nutrition is the most important factor or liver disease or a history of cancer. that regulates gut microbiota composition. Inclusion criteria for atherosclerosis were Personalized nutrition is one of the most [21]: patients with a BMI in the range of normal effective approaches for prevention and weight; low cardiovascular risk (SCORE 1%); treatment of NCDs [18]. The edible plants’ total cholesterol level below 8 mmol/l; total 88 Experimental articles triglycerides levels below 2.3 mmol/l; signed fold serial dilution of samples was performed informed consent to participate in the study. in PBS and plated correspondingly on the Exclusion criteria: patients receiving lipid- following nutrient media: Mitis Salivarius lowering therapy (statins, ezetimibe, etc.) or Agar, Bile Esculin Agar, Mannitol Salt patients who do not meet the minimum period Agar, Endo Agar, Bismuth Sulphite Agar, of 3 months of discontinuation of therapy; HiCrome Clostridial Agar, Sabouraud the lipid profile outside the inclusion criteria; Dextrose Agar, Lactobacillus MRS Agar, diabetes mellitus; SCORE > 1%; proven Bifidobacterium Agar, Bacteroides bile secondary causes of dyslipidemia; presence of esculin agar, Propionibacter Isolation Agar, manifest cardiovascular system disease in the L.D. Esculin HiVegTM Agar (manufactured form of coronary artery disease, past stroke, by HiMedia Laboratories, India), UriSelect™ TIA, MI, etc.; presence of acute diseases, 4 Medium (Bio-Rad Laboratories, Inc, USA), chronic deterioration, or presence of infection, and Blaurock semi-liquid modified hepatic which may distort the laboratory parameters; medium (manufactured
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