Analysis Of Differences In Compositions Of Lonicera Japonica Thunb. And Lonicera Flos Based On Serum Chemistry And Multiomics

Lonicera japonica Thunb.,also known as Lonicera,was the dried flower bud or first bloom of the Lonicera genus of the Caprifoliaceae family.Lonicera flos included Lonicera confusa(Sweet)DC.,Lonicera hypoglauca Miq.,Lonicera macranthoides Hand.-Mazz.,and Lonicera fulvotomentosa Hsu et S.C.Cheng,the first flowering buds can be used as medicine.It was traditional Chinese medicinal material that was commonly used and in greater demand.Lonicera japonica Thunb.and Lonicera flos are widely used in medicine,food,cosmetics and health industry.Lonicera japonica Thunb.and Lonicera flos belong to the phenomenon of“one medicine with multiple bases”.Due to the same source of plant species,the difference in appearance and form of medicinal materials,and the obvious difference in sexual taste and function of the pharmacopoeia,there is no obvious difference between the Lonicera japonica Thunb.and Lonicera flos.Lonicera japonica Thunb.and Lonicera flos had mixed use in clinical application and promotion.In order to clarify their material basis and differences in drug efficacy,the following related studies had been carried out.In this study,Lonicera japonica Thunb.and Lonicera flos(Lonicera fulvotomentosa Hsu et S.C.Cheng)were used as the objects.UPLC/Q-TOF-MS analysis technology was used to analyze and identify the extracts and serum transitional components of Lonicera japonica Thunb.and Lonicera flos;Establish a lipopolysaccharide(LPS)-induced acute lung injury(ALI)disease model in rats,and pre-intragastric the water extract of Lonicera japonica Thunb.and Lonicera flos in rats,using transcriptome sequencing and microbial diversity analysis to study ALI rats.In order to reveal the differences in the efficacy components,the protection of ALI and its possible mechanism of Lonicera japonica Thunb.and Lonicera flos.The main results are as follows:1.Study on the chemical composition of Lonicera japonica Thunb.and Lonicera flosEstablished the in vitro composition analysis method of Lonicera japonica Thunb.and Lonicera flos to detect and analyze the extracts of them;There were total of 46 componentswere identified in the water extract of Lonicera japonica Thunb.and 57 components in the water extract of Lonicera flos(Lonicera fulvotomentosa Hsu et S.C.Cheng).The chemical constituents of them are mainly flavonoids,organic acids,alkaloids,iridoids,saponins and other types of compounds.In this study,a large number of glycoside compounds and iridoid derivatives were detected in Lonicera flos,and the number was greater than Lonicera japonica Thunb.,which is of great significance for the subsequent identification of Lonicera flos for pharmacology.2.Analysis of serum transitional components of Lonicera japonica Thunb.and Lonicera flosUnder the guidance of serum medicinal chemistry,the serum of rats after intragastric of Lonicera japonica Thunb.and Lonicera flos water extracts were analyzed,and the acetonitrile deproteinization method was selected as the serum treatment method.Fifteen blood transitional components were detected in the serum-containing of rat oral Lonicera japonica Thunb.extract,six prototype components and six metabolites were identified;Fifteen blood transitional components were detected in a drug-containing serum sample of oral extract of Lonicera japonicus in rats,there are twelve prototype components and one of metabolite were identified.The common components of them after absorbing were chlorogenic acid compounds.Comparing the peak areas,it was found that their content after blood infusion was different.This may be the difference between the two used in clinical drug treatment.In addition,dipsacoside B was also detected in the serum sample of Lonicera flos,which is consistent with the in vitro,but dipsacoside B was not detected in vitro and in vivo of Lonicera japonica Thunb.,indicating that this component was one of the characteristic components between Lonicera japonica Thunb.and Lonicera flos.At the same time,it may also be the fundamental difference between Lonicera japonica Thunb.and Lonicera flos for clinical treatment.3.Transcriptome analysis of the effects of Lonicera japonica Thunb.and Lonicera flos on ALI ratsThrough pathological observation,the rats in the Lonicera japonica Thunb.group and Lonicera flos group had less lung damage than the model group;ELISA measurement results showed that compared with the model group,The serum and BALF of TNF-?,IL-1?,IL-6,MDA and MPO content in lung tissue was significantly reduced in the Lonicera japonica Thunb.group and Lonicera flos group,the concentration of GSH-Px and SOD in lung tissue increased significantly;After sequencing quality control,a total of 133.68 Gb Clean Data was obtained,the Q30 base percentage of each sample was not less than 94.05%.The genes with differential expression were initially screened: 919 DEGs in Lonicera japonica Thunb.group and model group,504 DEGs in Lonicera flos and model group,800 DEGs in the Lonicera japonica Thunb.group and Lonicera flos group,of which 476 were up-regulated and 324 were down-regulated.Analysis of the KEGG pathway in DEGs through Kobas 3.0 software,mainly involved in inflammation,cancer,immune system,intestinal diseases and other related pathways.RT-q PCR verification results were consistent with RNA-seq expression patterns in lung tissues.MRNA expression of CCDN1 and KITA was down-regulated and IL-1R1,IL-6R,CXCL1 and NFKBIA was up-regulated in Lonicera japonica Thunb.,the m RNA expression of THBS1,TNFAIP3,IL-1R1,IL-6R,CXCL1 and NFKBIA was down-regulated,and CCDN1,KITA was up-regulate in Lonicera flos group,there by they can reduce the lung inflammation in ALI rats.4.Microbiome analysis of the effects of Lonicera japonica Thunb.and Lonicera flos on ALI ratsThrough microbial diversity analysis,it was found that the intestinal flora of ALI rats was dysregulated.Lactobacillus,Trichophyceae NK4A136 group,uncultivated bacteria Muribaculaceae,uncultivated bacteria Trichoderma and Escherichia-Shigella and other fungi had a high correlation with the occurrence and development of ALI.Lonicera japonica Thunb.and Lonicera flos may improve the level of inflammatory factors by increasing probiotic bacteria such as Akkermansia,Bacteroides and Bifidobacterium to affect ALI.In summary,the chemical composition of Lonicera japonica Thunb.and Lonicera flos were significantly different,but the phenolic acid components were their common main component;The transition components of Lonicera japonica Thunb.and Lonicera flos were significantly different after blood absorbed,this study provided data support for the research on the efficacy of Lonicera japonica Thunb.and Lonicera flos;It was found that Lonicera japonica Thunb.and Lonicera flos can reduce lung inflammation by regulating inflammationand repairing intestinal flora in ALI rats.The mechanism may be related to regulating TNF and NF-?B signaling pathways,increasing probiotic bacteria in the intestinal tract and reducing the abundance of harmful bacteria.The purpose of this study was to explain the differences in the substance basis and efficacy caused by the difference between the original Lonicera japonica Thunb.and Lonicera flos from the basic research data.Therefore,this study provided guidance for its clinical application and promotion.