![]() Amplicon sequence variants (ASVs) were aligned using the mafft plugin in QIIME2. Taxonomic assignment of 16S rRNA sequences was performed using a Silva 138 99% taxonomy classifier. The paired-end sequences were de-noised and merged using the DADA2 plugin within QIIME2. Then, the Quantitative Insights Into Microbial Ecology 2 version 2022.8 (QIIME2) pipeline was used to analyze the sequence reads. Then, paired-end 2 × 250 bp sequencing was performed by the Illumina NovaSeq 6000 platform. The bacterial 16S rRNA genes V4 region was amplified by polymerase chain reaction (PCR) using 515F (5’- GTGCCAGCMGCCGCGGTAA-3’) and 806R (5’- GGACTACHVGGGTWTCTAAT-3’) primers. Aberrant Crypt Foci Analysisīacterial genomic DNA was extracted from the feces of each group of rats using the TIANamp Stool DNA Kit (Tiangen, Beijing, China). At the end of the experiments, all rats were sacrificed, and colons were collected for ACF evaluation ( Figure 1A). Fecal specimens were naturally collected (week 14, non-invasive, and sampled repeatedly) and subjected to intestinal microbiota studies. The serum was subjected for the examination of pro-inflammatory cytokine concentration (IL-6, IL-1β, TNF-α) by ELISA kit (Thermo Fisher Scientific, Waltham, MA, USA ) following manufacturer’s instructions protocol. Blood was collected from the lateral tail vein of the rats in week 3 (after DMH administration), week 5 (after DSS-induced inflammation), and week 10 (after five weeks of PCE intervention). In week 5, rats were orally fed with 10% DMSO (group 2) and 0.1 or 1 g/kg body weight/day of PCE (groups 3 or 4, respectively) until week 15. At week 3, rats were then given 1% dextran sulfate sodium (DSS) daily instead of drinking water for one week. injected with 40 mg/kg body weight of dimethylhydrazine (DMH) at week 1 and week 2. In positive control (group 2) and experimental groups (groups 3 and 4), rats were s.c. In control groups (groups 1 and 5), rats received a subcutaneous ( s.c.) injection of 0.9% normal saline once a week for two weeks and were fed with 10% DMSO (group 1 negative control) or 1 g/kg body weight of PCE (group 5 PCE control) in weeks five to 15. However, the mechanisms of PCE on the microbiota, which are related to inflammation and inflammatory-induced colon cancer progression, need to be further investigated.Īfter 1 week of acclimatization, rats were randomly separated into five groups of six rats with comparable average weights (111.87 ± 2.09 g). Moreover, consumption of PCE could alter rat microbiota, which might be related to health benefits. These results demonstrate that the active components in PF seed residue showed a preventive effect on the aberrant colonic epithelial cell progression by modulating inflammatory microenvironments from the infiltrated macrophage or inflammatory response of aberrant cells. Additionally, PCE could either modulate the inflammation induced in murine macrophage cells by bacterial toxins or suppress the proliferation of cancer cell lines, which was induced by the inflammatory process. PCE at high dose exhibited a reduction in aberrant crypt foci (ACF) number (66.46%) and decreased pro-inflammatory cytokines compared to the DMH + DSS group ( p < 0.01). PCE 0.1 and 1 g/kg body weight were administered by oral gavage to rats after receiving dimethylhydrazine (DMH) with one week of dextran sulfate sodium (DSS) supplementation. This study aimed to investigate the chemoprotective action of PF seed residue crude ethanolic extract (PCE) on the inflammatory-induced promotion stage of rat colon carcinogenesis and cell culture models. Perilla frutescens (PF) seed residue is a waste from perilla oil production that still contains nutrients and phytochemicals.
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |