RESUMO
Transcriptome sequencing was used to obtain the differentially expressed genes of conjugated linoleic acid (CLA) on the basal diet of Royal chickens on intramuscular fat metabolism in related signal pathways and candidate genes that may play important roles by bioinformatics analysis, which could provide a further understanding of the molecular mechanism of CLA on intramuscular fat deposition. In this study, 55-day-old healthy Royal chickens were selected as experimental animals, and 0%, 1% and 2% of CLA were added to the basic diet. The pre-feeding period was 1 week and the normal feeding period was 6 weeks. The breast muscle tissues were collected for transcriptome sequencing, and the sequencing data were subjected for differential expression analysis, such as GO function and KEGG pathway enrichment analysis of differentially expressed genes. Differentially expressed genes related to breast muscle lipid metabolism were screened out, and qRT-PCR was used to verify differentially expressed genes. The results showed that a total of 1 065 differentially expressed genes were obtained, of which 703 were up-regulated genes and 362 were down-regulated genes. GO enrichment results show that differentially expressed genes are concentrated in cellular processes, single-biological processes, biological regulation and metabolic processes in biological processes. Enrichment analysis of KEGG signaling pathway shows that differentially expressed genes were significantly enriched in focal adhesion, unsaturated fatty acid biosynthesis, fatty acid biosynthesis, and steroid biosynthesis. Totally 11 candidate genes were found mainly related to intramuscular fat metabolism, including FADS1, FADS2, ELOVL5, ACOX2, SLC27A1, FABP5, LPL, LOC107050163, ENSGALG00000030996, ENSGALG00000005043 and ENSGALG00000048882. Six genes were randomly selected for qRT-PCR verification, and their relative expression changes were consistent with the sequencing results. This study screened the differentially expressed genes related to CLA affecting breast muscle lipid metabolism in Royal chickens, and analyzed 11 genes related to fat metabolism, laying the foundation for revealing the molecular mechanism of CLA regulating intramuscular fat deposition.
RESUMO
To establish a method for the content determination of indexes for measuring aconitic compounds contained in Shenfu injection, in order to provide basis for the evaluation of the curative effect of monkshood in Shenfu injection. The sample were purified and enriched with HF-LPME. ACQUITY UPLC BEH C18 column (2.1 mm x 50 mm, 1.7 microm) was adopted and eluted with a gradient program, with acetonitrile-10 mmol x L(-1) NH4HCO3 (pH 10) as the mobile phases. The flow rate was 0.45 mL x min(-1). The content was determined with ESI and MRM. The results showed that aconitine, hypaconitine and mesaconitine showed a good linear relationship, with r > 0.999, within the range of 0.1-100 ng x L(-1). The recoveries were detected to be 100.1%, 97.4%, 97.5%, with RSD being 1.2%, 1.1%, 1.5%, respectively. This method was used to prove the safety of Shenfu injection, and provide scientific basis for correct evaluation of curative effect of monkshood, as well as a reliable, simple and practical means for quality control of monkshood-containing Chinese materia medica preparations.