Lipids from stool
In this study, we investigated how positively and negatively-charged lipid extract from feces of standard chow-fed mice (NL), by a simple extraction method, impact cell functioning. MTT assays for assessing cell metabolic activity indicated that NL differently affects tumor-derived cell lines, reducing the viability of Hela and HepG2 cells but it does not affect PC3 cells. Viabilities of 3T3-L1 fibroblasts and MSCs were not affected by NL. Additionally, NL displayed partial PPAR γ agonist action on reporter gene assays and on differentiation of 3T3-L1 fibroblasts to adipocytes. Transcriptome analyses showed that NL suppresses inflammatory response via toll-like receptor 4 (TLR4) signaling and affects PPAR target gene expression. Lipidomic analyses revealed significant differences of positively and negatively-charged lipid content of CD and NL, with NL containing less cancer correlated lipids and more apoptosis and necroptosis lipids than chow diet. These findings indicate that NL affect important aspects of cellular function and, therefore, may be considered as a promising therapeutic alternative to fecal transplant (FT).
Steps to reproduce
Lipids extraction from NL and chow diet (CD) were performed with methanol and chloroform. Cellular viability was tested in different cell lines as HeLa, HepG2, mesenchymal stem cells (MSC), PC3 and 3T3-L1 cells. To observe proliferator-activated receptor (PPAR) γ activity, gene reporter assay and 3T3-L1 differentiation into adipocytes, including RT-qPCR for differentiation genes. RNA-seq, were carried out to identify the genes affected by NL and a lipidomic were performed to show differences between extracts.