Mechanism of Anti-tumor Effects of Viola Medicinal Materials Based on Network Pharmacology and Molecular Docking

[Objectives]To explore the relationship between anti-tumor components,targets,and pathways involved in Viola medicinal materials,study its main active components,and evaluate its inhibitory activity.[Methods]Through network pharmacological analysis,molecular docking simulation experiments and in vitro experiments,the main components and corresponding targets of Viola were screened out,and their anti-tumor signaling pathways were confirmed.MTT colorimetric assay was used to investigate the inhibitory effect of different extraction layers of Viola on the growth of tumor cells.[Results]18 anti-tumor active components such as 2α,19α-Dihydroxyursolic acid,Corlumine,Madolin U,Trifolirhizin and etc.,and 52 action targets such as PTGS2,PTGS1,P2RX7,MMP1,and GABRB3,and anti-tumor signaling pathways were confirmed.The results of molecular docking showed that all the selected Viola compounds had good binding activity.The results of MTT colorimetric assay showed that the petroleum ether layer and n-butanol layer had a good inhibitory effect on the growth of tumor cell lines.[Conclusions]Viola medicinal materials have the potential of anti-tumor,triterpenoids and flavonoids may be the main active components,and the petroleum ether layer and n-butanol layer have better inhibitory effect on the growth of tumor cells.

Transcriptome profiling and RXR gene family identification reveals the molecular mechanism of rapid aging after spawning of cuttlefish Sepiella japonica

Sepiella japonica is a worldwide marine cuttlefish species of high economic value.S.japonica routinely modifying behaviors in reproductive life,such as rapid aging until death after spawning,has been recognized in artificial breeding.However,reproductive behavior at the level of genes is rarely reported,thus,the research on the genetic basis of behavior,reproduction,and artificial breeding was limited.We applied RNA-seq in different stages of reproduction to investigate the reason of rapid aging after spawning,pre-maturity,pre-spawning after maturity,and post-spawning.The retinoid X receptor(RXR)gene family in S.japonica was identified,and 1343–1452 differentially expressed genes(DEGs)in all 3 stages of reproductive life were identified from pairwise m RNA comparisons.Furthermore,through the GO term and KEGG analysis,S.japonica could handle neuronal development and network formation before maturity and have a functional degradation of neural communication,signal transduction,vision,and gene expression after spawning.Eight Sj RXRαs have been identified and they played different roles in growth development or reproduction.Therefore,the regulation of several channels and receptors is the intrinsic molecular mechanism of rapid aging after spawning in S.japonica.This study revealed the survival strategy and provided fundamental data on the level of genes for understanding the reproductive behavior and the reproduction of S.japonica.

Structural Characterization of Chloroplast Genome in Alpinia japonica(Thunb.)Miq.,a Medicinal Plant of the Genus Alpinia

The analysis of chloroplast gene characteristics in Alpinia japonica(Thunb.)Miq.is of great significance for developing relevant genetic resources.The high-throughput sequencing and bioinformatic research were performed to analyze the chloroplast genome characteristics of A.japonica.The total chloroplast genome length of A.japonica was 161,906 bp,with a typical circular tetrameric structure.And 133 genes were annotated,comprising 87 protein-coding,38 tRNA,and 8 rRNA genes.Furthermore,22 genes contained two copies,and 18 genes owned introns.Repeat sequence analysis showed that it contains 321 simple sequence repeats(SSRs)and 37 long segment repeats.Compared with the chloroplast genomes of eight representative plants in the genus Alpinia,the gene structure,type,and quantity were relatively conservative.Rps12 was the highest variation site in the entire chloroplast gene.A phylogenetic tree showed that the genus Alpinia was the most closely related to the genus Amomum.Meanwhile,A.japonica is the most closely related to Alpinia chinensis belonging to the genus Alpinia.Overall,the chloroplast genome of a new species was reported in the genus Alpinia,and a basis was provided for the utilization of Alpinia plants as a medical resource.

槐树(Sophora japonica L.)种质资源现状及研究进展

对槐树种质资源在我国的分布和现状,以及山东、山西、河北和河南等地的资源调查收集情况进行了综述;从表型、细胞和生理生化、分子水平3个方面,对槐树种质资源评价的研究进展进行了归纳;以选择育种、人工杂交授粉和现代生物技术育种为例,对种质资源创新利用情况进行了总结,并对工作中存在的问题和发展前景进行了分析。

Chemical constituents and pharmacological effects of Lonicerae japonicae

Lonicera japonica,also known as honeysuckle,is an evergreen shrub in the family of Syzygium.By consulting Sciencedirect databases and Web of Science databases,79 related articles were found,of which 22 were related to chemical composition and pharmacological activity.These articles show that L.japonica has a wide range of pharmacological activities,including antiviral,anti-tumor,and antioxidant effects.These activities have important applications in the pharmaceutical,food,and fragrance industries.This review focuses on the chemical composition and pharmacological effects of L.japonicae,which is of great significance to the development of new drugs and therapeutic methods.

Study on Skin Care Efficacy of the Active Ingredients in Camellia Japonica Flowers at High and Low Altitudes

The active ingredients of Camellia japonica flowers(CJF)at high and low altitudes,as well as their skin care efficacy were compared.The dried red CJF at high and low altitudes were ultrasonically extracted with 70%ethanol,and these extracts were concentrated and then diluted to a constant volume.The content of total flavonoids,total polyphenols and total proteins was tested and analyzed.In addition,DPPH free radical scavenging,inhibition of the formation of advanced glycation end products,and inhibitory activity against elastase was measured to compare their skin care efficacy in anti-oxidation,anti-glycation,anti-wrinkling and firming.The results showed that,based on the dry weight of CJF,the content of total flavonoids and total proteins of the CJF at high altitude was approximately 1.5 times of the CJF at low altitude,while the content of total polyphenols was approximately 2.4 times that of the CJF at low altitude.The skin care efficacy of CJF at high altitude was significantly better than that of the CJF at low altitude.This work could provide theoretical basis for the selection and application of Camellia japonica flowers in the field of cosmetics.

曼氏无针乌贼(Sepiella japonica)在不同盐度胁迫下的生理生化和组织学响应特征分析

盐度是影响海洋生物代谢和渗透调节最重要的生态因子之一,它的变化会影响头足类动物的能量收支、胚胎发育、个体生长、免疫能力、繁殖能力等多个方面,还会直接影响血清渗透压、离子浓度、抗氧化能力和游离氨基酸含量。为探究盐度胁迫对成体曼氏无针乌贼(Sepiella japonica)生理生化和组织学的影响,分别利用高盐(32)和低盐(18)对乌贼进行胁迫处理。结果显示,低盐胁迫6、12和24 h后,乌贼血淋巴中的Na^(+)、Cl^(-)、K^(+)、Ca^(2+)浓度相较于正常盐度组均显著性下降。高盐胁迫12和24 h后,血淋巴中的Na+浓度相较于正常盐度组显著性上升;Ca^(2+)浓度在12 h后显著性上升。肝脏中SOD活力和CAT的表达量在盐度胁迫6 h后显著下降。鳃中Na^(+)/K^(+)-ATP酶α亚基和Ca^(2+)-ATP酶的表达量在低盐胁迫24 h后显著下降。此外,H.E染色显示,低盐胁迫会造成乌贼鳃和肾组织核质比降低以及肝脏的病理性损伤,而在高盐胁迫后无变化。研究结果从生理生化水平和组织学水平解析曼氏无针乌贼成体在不同盐度胁迫下的响应特征,为曼氏无针乌贼和头足类的盐度响应研究提供了形态学支撑和理论依据,也对乌贼养殖过程中应对盐度波动具有一定的参考价值。

不同强度光强胁迫对海带(Saccharina japonica)幼苗光合生理的影响

海带(Saccharinajaponica)是我国藻类生产的主要品种之一,其栽培面积和产量均居世界首位。2021~2022年产季,山东荣成海带主产区先后暴发了大规模病烂灾害,给当地水产经济造成了巨大损失。引起病烂的原因可能来自多方面,其中藻体所受光照过强及水体营养盐水平较低可能起到了一定作用,为此,基于海带幼苗的最小饱和光强、烟台高新区牟平自然海区海水氮磷浓度及不同海水深度下的光照强度测定结果,研究了梯度强光在不同海水营养盐水平下对海带生理的影响,以期为“烂菜”现象的病因分析提供一定的线索。结果显示,海带幼苗在700~900μE/(m^(2) s)强光胁迫3 d后, Fv/Fm可恢复至对照水平,光合色素合成活跃,营养盐含量较高的海水更有助于PSII的修复,但1 300~1 500μE/(m^(2) s)的强光辐射导致PSII不能恢复,岩藻黄素、叶绿素a及β-胡萝卜素含量显著降低。整个胁迫实验中,藻体总抗氧化能力(T-AOC)及其他抗氧化酶比活力大体上在强光胁迫的起始至第3天上调,而在胁迫第5天时又出现下降趋势,且在营养盐含量较高的处理组中,各样本活性均高于天然海水处理组样本。抗氧化酶基因的表达也呈现类似的变化趋势。在低于900μE/(m^(2) s)的光照条件下,抗氧化酶活性可以得到较好的维持,而海水中相对较为丰富的营养盐则有利于藻体抗氧化能力的维持。研究结果为荣成海带病烂暴发的原因分析提供了一定的借鉴。

中国近海洄游日本鳗鲡(Anguilla japonica)玻璃鳗肠道微生物菌群分析

为研究长江口日本鳗鲡玻璃鳗肠道菌群的结构与多样性特征,初步探究野生鳗鲡仔鱼的营养需求,分别采集浙江台州市、舟山市以及江苏盐城市邻近海域的玻璃鳗样品,利用16S rDNA扩增子测序技术,分析三个采样点玻璃鳗肠道优势菌群及营养代谢相关的功能差异。结果显示,三个采样点的玻璃鳗肠道菌群均以变形菌门(Proteobacteria,68.7%~82.0%)和放线菌门(Actinobacteria,4.2%~19.0%)为主要优势菌门。在属分类水平上,不动杆菌属(Acinetobacter,12.4%~17.3%)和假单胞菌属(Pseudomonas,8.07%~13.03%)丰度较高,属固定优势菌属。此外,每个采样点样本中都含有丰富的独特细菌群,包括台州市的气单胞菌属(Aeromonas)、舟山市的内生单胞属(Endozoicomonas)和丙酸杆菌属(Cutibacterium)以及盐城市的嗜冷杆菌属(Psychrobacter)和稳杆菌属(Empedobacter),推测是受采样点不同水环境因素影响。不动杆菌、假单胞菌、内生单胞菌等菌属在多种鱼类肠道中有着消化蛋白质的功能,气单胞菌虽属于纤维素降解菌,也可分泌胞外蛋白酶。此外,通过对玻璃鳗肠道菌群进行功能预测发现,三个采样点玻璃鳗菌群功能均以氨基酸的转运和代谢占比最高,推测玻璃鳗在洄游过程中可能摄取较多的蛋白质成分作为其饵料。初步探明了我国近海野生玻璃鳗肠道微生物菌群的多样性组成,可为研究野生鳗鲡仔鱼摄食习惯及食物组成提供依据,同时为日本鳗鲡人工繁殖仔稚鱼的饵料设计提供参考。

Effects of Different Arbuscular Mycorrhizal Fungi on Physiology of Viola prionantha under Salt Stress

Arbuscular mycorrhizal(AM)fungi distribute widely in natural habits and play a variety of ecological functions.In order to test the physiological response to salt stress mediated by different AM fungi,Viola prionantha was selected as the host,the dominant AM fungus in the rhizosphere of V.philippica growing in Songnen saline-alkali grassland,Rhizophagus irregularis,and their mixtures were used as inoculants,and NaCl stress was applied after the roots were colonized.The results showed that V.philippica could be colonized by AM fungi in the field and the colonization rate ranged from 73.33%to 96.67%,and Claroideoglomus etunicatum was identified as the dominant AM fungi species in the rhizosphere of V.philippica by morphology combined with sequencing for AM fungal AML1/AML2 target.Inoculation with both the species resulted in the formation of mycorrhizal symbiosis(the colonization rate was more than 70%)and AM fungi significantly enhanced plants’tolerance to salt stress of varying magnitude.Higher activity of antioxidant enzymes and augmented levels of proline and other osmoregulators were observed in AM plants.The content of MDA in CK was higher than that in the inoculations with the stress of 100,200,and 250 mM.All indices except soluble protein content and MDA content were significantly correlated with AM fungal colonization indices.The analysis for different AM fungal effects showed that the mixtures and R.irregularis worked even better than C.etunicatum.These results will provide theoretical support for the exploration and screening of salt-tolerant AM fungi species and also for the application of AM-ornamental plants in saline-alkali urban greening.

Screening hepatoprotective effective components of Lonicerae japonica Flos based on the spectrum-effect relationship and its mechanism exploring

Lonicerae japonicae Flos(LF)is a kind of healthcare food with hepatoprotective function.This study was designed to explore the spectrum-effect relationships between UPLC fingerprints and the hepatoprotective effects of LF.Fingerprints of ten batches of LF were established by UPLC-PDA.The inhibitory levels of AST and ALT were used as pharmacological indexes,and secoxyloganin,isochlorogenic acid A and isochlorogenic acid C were screened as hepatoprotective active compounds by grey relational analysis(GRA)and partial least squares regression analysis(PLSR).Caspase-3 was obtained by network pharmacology as a key target of hepatoprotective active compounds.Molecular docking is used to explore the interaction between small molecules and proteins.This work provided a general model of the combination of UPLC-PDA and hepatoprotective effect to study the spectrum-effect relationship of LF,which can be used to considerable methods and insight for the fundamental research of the material basis of similar healthcare food.

Selection and Validation of Reference Genes for Normalization of RT-qPCR Analysis in Developing or Abiotic-Stressed Tissues of Loquat (Eriobotrya japonica)

Loquat(Eriobotrya japonica Lindl.)is a subtropical evergreen fruit tree that produces fruits with abundant nutrients and medicinal components.Confirming suitable reference genes for a set of loquat samples before qRT-PCR experiments is essential for the accurate quantification of gene expression.In this study,eight candidate reference genes were selected from our previously published RNA-seq data,and primers for each candidate reference gene were designed and evaluated.The Cq values of the candidate reference genes were calculated by RT-qPCR in 31 different loquat samples,including 12 subgroups of developing or abiotic-stressed tissues.Different combinations of stable reference genes were screened according to a comprehensive rank,which was synthesized from the results of four algorithms,including the geNorm,NormFinder,BestKeeper andΔCt methods.The screened reference genes were verified by normalizing EjLGA1 in each subgroup.The obtained suitable combinations of reference genes for accurate normalization were GAPDH,EF1αand ACT for floral development;GAPDH,UBCE and ACT for fruit setting;EF1α,GAPDH and eIF2B for fruit ripening;ACT,EF1αand UBCE for leaves under heat stress;eIF2B,UBCE and EF1αfor leaves under freezing stress;EF1α,TUA and UBCE for leaves under salt stress;ACT,EF1αand eIF2B for immature pulp under freezing stress;ACT,UBCE and eIF2B for immature seeds under freezing stress;EF1α,eIF2B and UBCE for both immature pulp and seeds under freezing stress;UBCE,TUB and TUA for red-fleshed fruits under cold-storage stress;eIF2B,RPS3 and TUB for white-fleshed fruits under coldstorage stress;and eIF2B,UBCE and RPS3 for both red-and white-fleshed fruits under cold-storage stress.This study obtained different combinations of stable reference genes for accurate normalization in twelve subgroups of developing or abiotic-stressed tissues in loquat.To our knowledge,this is the first report to obtain stable reference genes for normalizing gene expression of abiotic-stressed tissues in E.japonica.The use of the three most stable reference genes could increase the reliability of future quantification experiments.

Grain yield,nitrogen use efficiency and physiological performance of indica/japonica hybrid rice in response to various nitrogen rates

Utilizing the heterosis of indica/japonica hybrid rice(IJHR)is an effective way to further increase rice grain yield.Rational application of nitrogen(N)fertilizer plays a very important role in using the heterosis of IJHR to achieve its great yield potential.However,the responses of the grain yield and N utilization of IJHR to N application rates and the underlying physiological mechanism remain elusive.The purpose of this study was to clarify these issues.Three rice cultivars currently used in rice production,an IJHR cultivar Yongyou 2640(YY2640),a japonica cultivar Lianjing 7(LJ-7)and an indica cultivar Yangdao 6(YD-6),were grown in the field with six N rates(0,100,200,300,400,and 500 kg ha^(-1))in 2018 and 2019.The results showed that with the increase in N application rates,the grain yield of each test cultivar increased at first and then decreased,and the highest grain yield was at the N rate of 400 kg ha^(-1)for YY2640,with a grain yield of 13.4 t ha^(-1),and at 300 kg ha^(-1)for LJ-7 and YD-6,with grain yields of 9.4–10.6 t ha^(-1).The grain yield and N use efficiency(NUE)of YY2640 were higher than those of LJ-7 or YD-6 at the same N rate,especially at the higher N rates.When compared with LJ-7 or YD-6,YY2640 exhibited better physiological traits,including greater root oxidation activity and leaf photosynthetic rate,higher cytokinin content in the roots and leaves,and more remobilization of assimilates from the stem to the grain during grain filling.The results suggest that IJHR could attain both higher grain yield and higher NUE than inbred rice at either low or high N application rates.Improved shoot and root traits of the IJHR contribute to its higher grain yield and NUE,and a higher content of cytokinins in the IJHR plants plays a vital role in their responses to N application rates and also benefits other physiological processes.

不同施肥处理对紫金牛(Ardisia japonica)生长和生理特性的影响

氮、磷、钾施肥对紫金牛(Ardisia japonica)的生长及生理特性产生明显影响,但是最佳施肥比例缺少研究报道。论文根据不同施肥处理对紫金牛的生长及生理的影响,筛选出适宜的施肥组合,为紫金牛苗木的施肥管理提供指导。以三年生紫金牛苗木为试验材料,采用三因素四水平的正交试验设计方法。结果表明:不同施肥处理能显著促进紫金牛苗木的生长,提高碳氮代谢产物的含量和叶绿素的含量,显著提升净光合速率。氮肥对紫金牛茎和叶的生物量、可溶性蛋白含量、叶绿素含量、净光合速率的影响达到显著性水平(P<0.05,下同);磷肥对紫金牛叶绿素含量的影响达到显著性水平;钾肥对紫金牛可溶性糖含量的影响达到显著水平。综合分析紫金牛生长指标及生理指标得出最适合的施肥量为尿素1.0 g·株^(-1)、过磷酸钙1.0 g·株^(-1)、硫酸钾1.5 g·株^(-1),能提高紫金牛的观赏性和加速其生长。

日本结缕草(Zoysia japonica)酵母杂交cDNA文库的构建及分析

酵母双杂交和单杂交技术是生物大分子互作和调控研究中的高效分子生物学技术,是蛋白质互作或蛋白质与DNA互作筛选的手段。本研究旨在构建高容量的日本结缕草(Zoysia japonica)酵母文库,以便后续深入挖掘日本结缕草相关基因,进行基因功能及基因网络研究。本研究应用不同组织来源以及不同生长时期的日本结缕草,通过SMART技术构建了高容量的酵母杂交cDNA文库。质量检测结果显示:次级文库库容为4.0×10^(6)cfu·mL^(-1),总克隆数为1.6×10^(7),重组率>95%。最终获得的Y187酵母文库工作液细胞密度为3×10^(7)cells·mL^(-1),插入片段具有良好的多态性。以日本结缕草类胡萝卜素裂解双加氧酶ZjCCD7作为诱饵,从构建的酵母文库中筛选到一个与ZjCCD7互作的蛋白ZjatpG,证明该文库质量较高,符合酵母杂交筛选试验标准,可适用于调控和互作蛋白筛选。本研究为筛选日本结缕草的互作蛋白以及基因功能研究提供了基础。

Anti-inflammatory maistemonine-class alkaloids of Stemona japonica

Three hitherto undescribed Stemona alkaloids,named stemajapines A-C(1-3),along with six known alkaloids(4-9),were isolated and identified from the roots of Stemona japonica(Blume)Miq.(Stemonaceae).Their structures were established by the analysis of the mass data,NMR spectra,and computational chemistry.Stemjapines A and B were degraded maistemonines without spiro-lactone ring and skeletal methyl from maistemonine.Concurrence of alkaloids 1 and 2 revealed an undescribed way to form diverse Stemona alkaloids.Bioassay results disclosed the anti-inflammatory natural constituents stemjapines A and C with IC_(50) values of 19.7 and 13.8μM,respectively,compared to positive control dexamethasone with 11.7μM.The findings may point out a new direction of Stemona alkaloids inaddition to its traditional antitussive and insecticide activities.

Identification and Characterization of Long Non-Coding RNAs Involved in Sex-Related Gene Regulation in Kelp Saccharina japonica

Long non-coding RNAs(lncRNAs)regulate a variety of biological processes,including sexual reproduction and differentiation.Saccharina japonica,a commercially important brown alga in China,shows remarkable sexual dimorphism in haploid gametophytes.The sex of Saccharina japonica gametophytes is determined by UV sexual system.However,no results have been reported on the lncRNAs involved in the sex-related gene regulation of S.japonica.This study identified a number of lncRNAs and assessed their expression levels in male and female gametophytes.Among them,a total of 405 lncRNAs and 211 mRNAs showed differential expressions.Furthermore,the functions of target genes of differentially expressed lncRNAs(DELs)differed from those of differentially expressed genes(DEGs),suggesting that lncRNA may interact with other functional proteins,in addition to DEGs,to involve sex regulation in S.japonica.There were 32 and 90 potential cis-regulatory and trans-regulatory interactions between DELsDEGs,respectively.Five of these lncRNAs(LNC_002974,LNC_021059,LNC_038466,LNC_051584,and LNC_027400)interacted with putative male sex determination region(SDR)genes,suggesting that they act as regulators in gametophytes'sex regulation potentially.Findings from this study contribute to our understanding of the roles of lncRNAs in sex differentiation and lay the foundation for functional studies of candidate lncRNAs in the future.

Regulation of Reproduction in Delayed Gametophyte of Saccharina japonica (Laminariales, Phaeophyceae): Effects of Light Intensity, Quality and Photoperiod

Saccharina japonica gametophytes can survive a long period under unfavorable environmental conditions,while they also delay in growth and/or reproduction.Although the reproduction in delayed gametophyte of S.japonica was known to be strongly influenced by light intensity,light quality,and photoperiod,no previous studies have evaluated their interactive effects on gametogenesis.To evaluate these effects,we used an orthogonal experiment to expose delayed gametophytes of S.japonica to different light intensities,light qualities,and photoperiods for 12 days.The results showed that changes in light intensity rather than light quality and photoperiod significantly affected the relative growth rates of the delayed gametophytes.Blue light had the greatest promotion on reproduction rate.The optimal light conditions in the early vegetative growth phase in gametogenesis induction for the delayed gametophytes were at 60–80μmol photons m^(−2) s^(−1) with daylength of 12 or 16 hours under white or blue light.When the delayed gametophytes were maintained in a constant light condition from delayed state to gametogenesis,the beneficial photoperiods for vegetative growth and reproductive rate were both 16L(16 hours of light):8D(8 hours of dark).However,when the delayed S.japonica gametophytes achieve the optimal growth state during the first 6 days and then they were cultured at different light conditions for the following 6 days,the reproduction rate increased as the daylength decreased and attained a peak value in group of 8L:16D photoperiod,indicating that photoperiod adjustment at the transition period is crucial in the gametogenesis induction process of delayed gametophyte of S.japonica.

Study on Tissue Culture of Japanese Honeysuckle ( Lonicera japonica )

[Objectives]This study was conducted to increase the reproduction coefficient of Japanese honeysuckle(Lonicera japonica)to keep the character of optimal benign.[Methods]The young leaves of medicinal Japanese honeysuckle were selected as explants,and MS was used as the basic culture medium.Suitable culture concentrations and conditions were screened through different concentration gradients of growth regulators and cytokinin.[Results]As the concentration of 6-BA in the culture medium increased,the browning rate increased,and the browning phenomenon occurred earlier.On the contrary,a lower concentration of 6-BA was suitable for the differentiation and growth of young leaves,and the browning response was slow.However,if the cultivation time was too long and the materials were not transferred in a timely manner,browning would also occur.The optimal combination of levels was obtained through a 3×3 orthogonal experiment(three parallel groups for each of 6-BA and NAA).The culture conditions included a constant temperature of 26℃and light intensity of 1200 lx.The optimal medium for inducing callus proliferation was MS+6-BA 0.5 mg/L+NAA 0.5 mg/L;and the optimal medium for inducing bud differentiation was MS+6-BA 1.0 mg/L+NAA 0.1 mg/L.[Conclusions]This study provides a theoretical basis for accelerating the development of the honeysuckle industry.

Chemical components and protective effects of Atractylodes japonica Koidz.ex Kitam against acetic acid-induced gastric ulcer in rats

BACKGROUND Atractylodes japonica Koidz.ex Kitam.(A.japonica,Chinese name:Guan-Cangzhu,Japanese name:Byaku-jutsu),a perennial herb,which is mainly distributed in northeast area of China,it’s often used to treat digestive system diseases such as gastric ulcer(GU).However,the mechanism of its potential protective effects against GU remains unclear.AIM To investigate the protective effects of A.japonica on acetic acid-induced GU rats.METHODS The chemical constituents of A.japonica were determined by ultra performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS)analysis.The rat model of GU was simulated by acetic acid method.The pathological changes of gastric tissues were evaluated by hematoxylin-eosin stain,the levels of epidermal growth factor(EGF),EGF receptor(EGFR),nuclear factor kappa-B(NF-κB),interleukin-1β(IL-1β),IL-10,Na^(+)-K^(+)-ATPase(NKA)in serum and gastric tissues were determined by enzyme-linked immunosorbent assay,and the mRNA expressions of EGFR,NF-κBp65,IkappaBalpha(IκBα)and Zonula Occludens-1(ZO-1)in gastric tissues were determined by real-time reverse transcription polymerase chain reaction,and the efficacy was observed.Then,plasma metabolomic analysis was performed by UPLC-MS/MS to screen the specific potential biomarkers,metabolic pathways and to explore the possible mechani-sms.RESULTS 48 chemical constituents were identified.Many of them have strong pharmacological activity,the results also revealed that A.japonica significantly improved the pathological damage of gastric tissues,increased the expression levels of IL-10,IκBαrelated to anti-inflammatory factors,decreased the expression levels of IL-1β,NF-κB,NF-κBp65,related to proinflammatory factors,restored the levels of factors about EGF,EGFR,ZO-1 associated with ulcer healing and the levels of factors about NKA associated with energy metabolism.Metabolomic analysis identified 10 potential differential metabolites and enriched 7 related metabolic pathways.CONCLUSION These findings contribute to the understanding of the potential mechanism of A.japonica to improve acetic acidinduced GU,and will be of great importance for the development and clinical application of natural drugs related to A.japonica.