Metagenomic DNA profile of gut microbioma in patients with rosacea

Introduction and Aim: The introduction of the new generation of sequence analysis systems allows for better understanding of the importance of the microbiome, as it is possible to detect almost all of the intestinal microbiome. In the studies, intestinal microbiome is associated with many diseases such as diabetes, hypertension and depression. This advanced technology, which is also used for other body areas, has also been used to demonstrate the pathogenesis of some dermatological diseases such as acne vulgaris, psoriasis and atopic dermatitis. Dermatological disease mechanisms are not only affected by skin microbiome but also by intestinal microbiome. The pathophysiology of rosacea disease, a chronic inflammatory dermatosis, has not been fully elucidated and its diagnosis can be made clinically. The aim of this study is; To determine the relationship between rosacea disease and intestinal microbiome, to shed light on the pathogenesis of rosacea and to help the new diagnosis and treatment approaches. Material and Method: 20 volunteers with rosacea with no known systemic and / or dermatological disease, aged between 18-49 years, who had not used antibiotics, proton pump inhibitors and / or probiotics, prebiotics for at least 4 weeks or less, and 10 healthy volunteers with the same conditions was included in the study. A mini questionnaire including questions not only about drug use but also about diet, smoking and alcohol use was applied to the volunteers. Fresh stool samples were taken from patients in samples vessels under sterile hospital conditions. All samples were applied Bristol stool scoring. After the pH measurements of the collected stool samples (Merck, NJ, USA), approximately 1 g stool samples taken from the patient and control groups were placed in the stabilization buffer (DNA / RNA Shield ™ Collection Tube, Zymo Research, CA, USA) at -20°C for a few days. Nucleic acid isolation (ZymoBIOMICS DNA Mini kit, Zymo Research, CA, USA) was then performed. Bacterial 16S ribosomal RNA (rRNA) gene target sequencing was performed using the 341f (CCTACGGGNGGCWGCAG) and 805r (GACTACHVGGGTATCTAATCC), targeting the V3-V4 region of the 16S rRNA gene of the universal bacteria 16S primers. The final library was created with Illumina MiSeq. Taxonomical maps were prepared by hierarchical clustering of taxa and samples by making appropriate amplicon array definitions and regulations. After bioinformatics, statistical analyzes were performed (LefSe). With the default settings (p <0.05 and LDA effect size> 2) taxa which are significantly different were determined between groups. Results: Patients with rosacea were found to have higher stool pH averages and Bristol scores than healthy volunteers in the control group. According to the results of 16S rRNA sequence analysis, in patients with rosacea, there was low alpha diversity and there was difference in beta diversity compared to healthy control group. According to the results of metagenomic DNA analysis; in rosacea patients; Lachnospira, Lachnoclostridium, Roseburia and Roseburia intestinalis were found at a higher rate, in healthy volunteers Prevotellaceae, Prevotella copri, Coriobacteriaceae, Desulfovibrio, Ruminococcaceae, Ruminococcus, Clostridiales, Clostridiales Family 13, Selenomonadales, Succinivibrio, Hungatella, Mollicutes, Butyricimonas virosa, Alisitipes, Oscillospira and Erysipelotrichaceae, factors on the basis of genus, species, family and order, were found to be statistically higher. Conclusion: There was a relationship between rosacea disease and intestinal dysbiosis. In this study, in rosacea patients according to healthy group; Lachnospira, Lachnoclostridium, Roseburia and Roseburia intestinalis at a higher rate. However, in the literature there is a need for more studies with more and more sampling in order to make sense of the microorganisms that stand out because there are very few studies done with rosacea and intestinal microbiome (only one study has been achieved). Keywords: Rosacea, gut microbiome, 16S rRNA, Roseburia, next generation sequencing