ABSTRACT
In the latest research, the concept of stress is associated with the deregulation of several biological systems sensitive to stress, such as the immune system, the microbiome, the endocrine system and neuroanatomical substrates. The objective of the research was to identify the fecal microbiome in patients diagnosed with chronic stress and in healthy patients through a metabarcoding analysis. The methodology used fecal samples collected from 20 patients with stress and 20 healthy patients. For the diagnosis of stress, psychological tools previously validated by external researchers were used. For metabarcoding analysis, metagenomic DNA extraction was performed from the fecal samples. Next Generation Illumina genetic sequencing targeting the 16S rDNA gene was then performed, followed by bioinformatic analysis using QUIME II software. The results, at the psychological test level, 20 people with chronic stress were diagnosed, on the other hand, at the metabarcoding level, specifically at the Gender level, the Asteroleplasma bacteria present only in the 20 healthy patients was molecularly identified. On the other hand, the bacteria Alistipes and Bifidobacterium were identified with greater predominance in the 20 patients with stress. Concluding, the bacteria Alistipes and Bifidobacterium are candidates as possible markers of the intestinal microbiome in patients with chronic stress, and the bacteria Asteroleplasma are candidates as a bacterial marker of the intestinal microbiome in healthy people. Finally, the identification of the microbiome in patients with stress opens a new path to understanding stress and its relationship to dysregulation with the microbiome.
ABSTRACT
Hepatic fibrosis is caused by chronic injury due to toxic, infectious, or metabolic causes, and it may progress to cirrhosis and hepatocellular carcinoma. There is currently no antifibrotic therapy authorized for human use; however, there are promising studies using cell therapies. There are also no animal models that exactly reproduce human liver fibrosis that can be used to better understand the mechanisms of its regression and identify new targets for treatment and therapeutic approaches. On the other hand, mesenchymal stem cells (MSC) have experimentally demonstrated fibrosis regression effects, but it is necessary to have an animal model of advanced liver fibrosis to evaluate the effect of these cells. The aim of this work was to establish a protocol for the induction of advanced liver fibrosis in rats using thioacetamide (TAA), which will allow us to perform trials using MSC as a possible therapy for fibrosis regression. For this purpose, we selected 24 female rats and grouped them into three experimental groups: the control group (G-I) without treatment and groups II (G-II) and III (G-III) that received TAA by intraperitoneal injection for 24 weeks. Then, 1 × 106/kg adipose mesenchymal stem cells (ASCs) were infused intravenously. Groups G-I and G-II were sacrificed 7 days after the last dose of ASC, and G-III was sacrificed 8 weeks after the last ASC infusion, all with xylazine/ketamine (40 mg/kg). The protocol used in this work established a model of advanced hepatic fibrosis as corroborated by METAVIR tests of the histological lesions; by the high levels of the markers α-SMA, CD68, and collagen type I; by functional alterations due to elevated markers of the hepatic lesions; and by alterations of the leukocytes, lymphocytes, and platelets. Finally, transplanted cells in the fibrous liver were detected. We conclude that TAA applied using the protocol introduced in this study induces a good model of advanced liver fibrosis in rats.
