Based On RNA-Seq To Analyze The Effect Of Alkaloids Of Sophorea Alopecuroides On Sulfur Metabolic Pathways And ROS Protection In Staphylococcus Epidermidis

Staphylococcus epidermidis is an opportunistic pathogen,once it formed the biofilm in vivo,it will become a repository of resistant gene,causing serious resistance problems.The latest research shows that traditional Chinese medicine can reverse the drug resistance,and enhance the antibacterial effect of existing antibiotics,but the mechanism of traditional Chinese medicine is very complex.Decipher the mechanism of traditional Chinese medicine in order to expand the clinical application of traditional Chinese medicine..The standard strain ATCC35984 and isolated strain S3 were used as the research targets.The expression of differentially expressed genes was analyzed by bioinformatics method after transcriptome sequencing technique.Real-time fluorescence quantitative PCR was used to verify the level of gene expression in sulfur metabolism pathway.The result showed that the biofilm formation ability decreased after using drugs for 12-18 h.The differential expression gene analysis showed that the proportion of antioxidant activity was higher,indicating that the expression of this type of gene was more.The expression of differentially expressed genes in the sulfur metabolic pathway was the highest,and all of them were down-regulated,indicating that Alkaloids of sophorea alopecuroides(TASA)can affect cysteine synthase,serine acetyltransferase,sulfite reductase and other key enzymes,and affect the whole sulfur metabolic pathway.The changes of reactive oxygen species in bacteria were detected after using drugs.The nitric oxide(NO)and enzyme activities of antioxidant system were detected by kits.The results showed that the production of ROS was stimulated by MIC and 1/2MIC,which was 2-4 times higher than that of ROS in the pre-treated cells,and the content of ROS in the cells showed a certain concentration-dependent.It can be seen that ROS-dependent antibacterial mechanism is a common antibacterial mechanism.The changes of MIC and ROS levels were detected after using drugs with sub-inhibitory concentration.Sum up the relationship between drug-induced ROS and drug resistance.Studies have found that antibiotics and traditional Chinese medicine could stimulate the ROS with the sub-inhibitory concentration.The yield of ROS decreased while the MIC increased with the increase of the times,and the MIC and ROS of the sub-inhibitory concentration of Chinese herbs were not changed in different generations.It is suggested that antibiotic-induced ROS helped to sterilization at high concentration,and ROS was conducive to bacterial resistance at low concentration,while traditional Chinese medicine was not easy to produce resistance,and could improve the sensitivity of antibiotics to bacteria.Combination of Chinese medicine and antibiotic could also effectively alleviate the bacterial resistance to antibiotic.