In order to investigate the significance of the pairing of Radix Bupleurum(RB)and Radix Paeoniae Alba(RP),the optimal extraction conditions were determined by single-factor investigation,and the volatile components of RB,RP and RB-RP were determined by headspace solid-phase microextraction(HS-SPME)coupled with gas chromatography-mass spec-trometry(GC-MS),and the percentage content of each component was determined by area normalization and subjected to principal component analysis(PCA).The results showed that there were ten volatile components shared by RB,RP and RB-RP,but the contents of the shared components were different.The PCA scores of the three pairs were significantly different,with the highest PCA score for the volatile components of the pair.Most of the volatile components of the pair of RB-RP came from RB,and the combination of RB-RP produced new active ingredients,and from the volatility point of view,the combina-tion of RB-RP has certain potency and toxicity reduction effects.
In order to promote the efficient enrichment of huperzine A,a new co-culture system(CCS),utilizing the symbiotic relationship between Huperzia serrata and endophytic fungi,was proposed.The optimal culture conditions of the CCS were ob-tained based on the cell survival rate,callus induction rate and multiplication rate.Huperzine A and ergosterol were applied as indicators to control the proportion of botanical cells and endophytic fungi.The growth balance between botanical cells and en-dophytic fungi was maintained for the optimal time of subculturing.The results showed that the best culture conditions for CCS was:100 Lux Luminous level,95％humidity,MS liquid medium,20.0 g/L sucrose,5.0 mg/L 2,4-dichlorophenoxyacetic acid and 1.0 mg/L kinetin.The 4th day was the optimal subculturing time with the highest concentration of huperzine A,which increased by 49.34％compared with the huperzine A level on the 1stday.The growth of endophytic fungal had a posi-tive effect on the enrichment of huperzine A when the concentration ratio of huperzine A to ergosterol was kept between 1.5 and 2.2.This study provides a new proposal to increase the production of huperzine A with high efficiency and low cost,which can effectively solve the problem of insufficient drug source and has a good application prospect.
This study aimed to investigate the variety and distribution of 7H-azulene[1,2,3-i,j]isoquinoline-7-one(7H-AI7O)components from Acorus traditional Chinese medicine.Using liquid-liquid extraction,solvent extraction,column chro-matography separation technology and thin-layer chromatography,the enrichment fragments of 7H-AI7O components from Acori Calami Rhizome and Acori Tatarinowii Rhizome were prepared with neo-tatarine as the reference substance,and then UPLC-Q-TOF/MS2 was used to analyze the composition of 7H-AI7O.Identified by thin-layer chromatography,among 10 bat-ches of Acorus traditional Chinese medicine,only batches S2,S4 and S5 of Acori Calami Rhizome and S6 of Acori Tatarinowii Rhizome contained 7H-AI7O components.According to the analysis of the mass spectrometry fragmentation rule of neo-tata-rine and target compounds,five 7H-AI7O compounds were detected in batch S2,two 7H-AI7O compounds were detected in batch S4,and only neo-tatarine was detected in batch S5 and S6.Although the number of test samples is limited,according to the analysis of the current results,there are 7H-AI7O components in both Acori Calami Rhizome and Acori Tatarinowii Rhizo-me,the distribution of the origin of Acori Calami Rhizome containing 7H-AI7O compounds is wider than that of Acori Tatari-nowii Rhizome,and types of 7H-AI7O components in Acori Calami Rhizome were more than in Acori Tatarinowii Rhizome.This study provides experimental basis for multiple plant sources,trace component detection,and related mechanisms of action of 7H-AI7O components.
To study the diversity and difference of rhizosphere microorganisms of Dendrobium nobile with different epiphytic substrates under the same cultivation environment,in order to clarify the influence of substrates on its rhizosphere microorgan-isms.The 16S and ITS sequencing of rhizosphere microorganisms of D.nobile in five epiphytic substrates,including stone and wood,using high flux sequencing technology.The diversity and difference of D.nobile rhizosphere microorganisms were com-pared by combining OTU analysis,Alpha diversity analysis and principal component analysis.A total of 56 phyla,112 classes,275 orders,377 families,and 621 genera of bacteria have been identified;Fungi:13 phyla,47 classes,123 orders,273 fami-lies,503 species.The dominant genus of D.nobile fungi in the stone matrix is unidentified_Cantharellales_Sp bacteria;the dominant bacteria are Sphingomonas,Fecal bacilli,and unidentified_Chloroplast;The dominant fungus genus of D.nobile in wood matrix is unidentified_Cantharellales_Sp bacteria,Entoloma,the dominant bacteria is Faecalibacterium.The relative a-bundance of D.nobile in the stone stroma was significantly different from that in the wood stroma,and the dominant genuses were different.The function prediction analysis showed that the epiphytic matrix was closely related to the function of D.nobile rhizosphere microbial community.The difference of rhizosphere microorganisms in different epiphytic substrates of D.nobile may affect the composition of its mycorrhizal fungi,thereby affecting the formation of quality differences in D.nobile.This study provides an experimental basis for the correlation of"substrate-rhizosphere microorganisms-mycorrhizal fungi"of D.nobile,its growth and development,and the accumulation of secondary metabolites,as well as the basis for"the Dendrobium grown on the stone has the superior quality"of D.nobile and the reasonable selection of cultivation substrate of D.nobile.
This study aims to study the constituents from the pericarp of Tamarindus indica Linn.and their antioxidant activi-ty.Eighteen compounds were isolated from the acetone extract by chromatographic separation technology.On the basis of NMR spectroscopic data,they were identifeid as luteolin(1),7,3',4'-trihydroxyflavone(2),apigenin(3),7,4'-dihydroxyflavone(4),geraldone(5),astragalin(6),quercetin(7),kaempferol(8),acacetin(9),eriodictyol(10),naringenin(11),butin(12),dihydrokaempferol(13),3',5,5',7-tetrahydroxyflavanone(14),5,6,7,4'-tetrahydroxyflavanone(15),3,3',5,5',7-pentahydroxyflavanone(16),catechin(17),epicatechin(18).Ten of them(2,4-6,9,11,13-16)were isolated from this ge-nus for the first time.By ORAC assay in vitro,it was found that four extracts and nine compounds(1,2,7-10,12,13,17)ex-hibited antioxidant activity.
Four natural products were isolated from the fermentation extract of the marine fungus Aspergillus jensenii SS5 using various column chromatographic separation techniques,and their structures were identified as epigriseofulvin(1),sterigmato-cystin(2),brevianamide M(3),and meleagrin(4)by NMR,HR-ESI-MS and X single-crystal diffraction.Compounds 1-3 were obtained for the first time,and all the compound were tested for in vitro cytotoxicity by SRB assay.The results of cytotox-ic activity test showed that compound 4 had inhibitory effect on human lung cancer A549 and human liver cancer Bel-7402,and compound 2 had inhibitory effect on A549.The study of the chemical composition of the marine fungus A.jensenii SS5 has some guiding significance,and two compounds were found to have good cytotoxic activity,which provides some theoretical ba-sis for the development of active natural products of marine fungi of the genus Aspergillus.
Lonicerae Flos is a valuable medicinal resource,but the chemical composition and biological activity of its oil have not been studied in depth.In this study,the Lonicerae Flos oil was prepared by supercritical CO2 extraction method.A system-atic characterization of chemical components was performed based on gas chromatography-mass spectrometry(GC-MS)and ultra-high performance liquid chromatography-high resolution mass spectrometry(UHPLC-QE-MS).Its biological activities such as anti-inflammatory and antioxidant activities in vitro were also studied.The GC-MS results based on non-methylation and methyl esterification methods indicated a total of 66 chemicals and the main compounds were fatty acids,sterols,and al-kanes.UHPLC-QE-MS identified 83 and 71 compounds in positive and negative ion modes,respectively,mainly including ter-penoids and flavonoids.Moreover,antioxidant activity analysis indicated that the ability of Lonicerae Flos oil to scavenge ABTS free radical has a good dose-dependent,and the IC50 of Lonicerae Flos oil involved in the DPPH free radical scavenging assay was 3.49 mg/mL.Meanwhile,The anti-inflammatory test showed that oil can significantly inhibit the expression of IL-6(P＜0.05)and NO(P＜0.05)levels in RAW 264.7 cells induced by LPS.These results show that the Lonicerae Flos oil is a natural active source with good anti-inflammatory and antioxidant activities,and it has good development and application value in Food and medicine.
To explore the effect of atractylenolide Ⅱ(AT-Ⅱ)on the polarization of M2 macrophages and the migration of A549 cells,and to analyze its potential mechanism.THP-1 cells were induced into M0 macrophages using PMA,and then IL-4 and IL-13 were added to continue inducing M0 macrophages into M2 macrophages.M2 macrophages were co-cultured with A549 cells in Transwell chambers and treated with 0,2.5,5 μmol/L AT-Ⅱ.MTT assay was used to determine the effect of AT-Ⅱ on the viability of A549 cells under co-culture condition;Real-time PCR was used to detect the mRNA expression level of Arg-1;ELISA was used to detect TNF-α,IL-1β content in co-culture system;WB was used to detect the protein expression levels of TLR4,p-p65,NF-κB(p65),PDL1;Scratch experiment was used to detect the migration ability of A549 cells.The results showed that under co-culture condition,compared with the control group,the activity of A549 cells in the experimental group decreased,2.5 μmol/L group(P＞0.05),5 μmol/L group(P＜0.01);The mRNA expression level of M2 macro-phage specific gene Arg-1 was significantly reduced(P＜0.01);The content of M1 macrophage associated inflammatory fac-tor TNF-α,IL-1β increased,but did not constitute a significant difference(P＞0.05);The expression levels of TLR4,p-p65 and NF-κB(p65)proteins related to the TLR4/NF-κB signaling pathway decreased in A549 cells(P＜0.01);The expres-sion of PDL1 protein in A549 cells significantly decreased(P＜0.01),and the ability of A549 cells migration decreased.These results indicate that AT-Ⅱ can reverse the polarization of M2 macrophages,inhibit NF-κB signaling pathway in A549 cells and reduce the expression level of PDL1,thus inhibiting the migration of lung cancer cells.
The pathogenesis of atherosclerosis(AS)is complex and is the pathological basis of various cardiovascular disea-ses.Autophagy is a catabolic mechanism of cells,and the dysfunction of autophagy is closely related to the occurrence and de-velopment of many human diseases.A growing body of research has revealed the potential of natural products,the collective name for the chemical components found in nature,to treat human diseases.Here,the recent studies on the improvement of AS by natural products based on autophagy are reviewed.
Mume Flos is the dried flower bud of Prunus mume(Sieb.)Sieb.et Zucc.in the Rosaceae family,and the Prunus mume"Lve"is the main species used in clinical medicine.In this study,the chemical composition of Mume Flos was deter-mined by ultra-high performance liquid chromatography with quadrupole electrostatic field orbitrap mass spectrometry(UPLC-QE-MS)at different developmental periods.A total of 328 compounds were detected,including 67 flavonoids,50 phenylpro-panoids,104 terpenoids,26 phenols,24 alkaloids,11 organic acids and their derivatives,8 amino acid derivatives and 38 oth-ers.The semi-quantitative results showed a gradual decrease in the content of the main chemical constituents in Mume Flos with the developmental period.The content of the total flavonoid and phenylpropanoid was determined by UV-Vis spectropho-tometry in Mume Flos samples at different developmental periods.The content of eight indicator components(neochlorogenic acid,chlorogenic acid,caffeic acid,rutin,hyperoside,isoquercitrin,isochlorogenic acid B and quercitrin)at different develop-mental periods was determined by UPLC,the results of the analysis showed significant differences in the content of compo-nents between the bud period(HL)and the full-opening period(QK)and that chlorogenic acid could be used as a marker of differential quality between the different developmental periods.The results of the DPPH radical scavenging assay on the antioxidant activity of Mume Flos at different developmental periods showed that the antioxidant capacity was closely related to the content of its components and gradually decreased with the development of flowers.The results of this paper can provide analytical methods and data support for the dynamics of the chemical constituents and the determination of the optimum har-vesting period of Mume Flos.
This study aimed to investigate the chemical constituents and antibacterial mechanism of Bletilla striata(Thunb.)Reichb.f.against Staphylococcus aureus.A comprehensive chromatographic separation was conducted to isolate and purify ac-tive compounds from B.striata,then all the isolated compounds were identified by NMR spectra.The inhibitory effect of the i-solated compounds on S.aureus was evaluated via broth microdilution.The antibacterial mechanism of the active ingredients in B.striata was explored from several aspects including the growth curves of S.aureus,extracellular alkaline phosphatase con-tent,DNA leakage,cell wall and cell membrane integrity.Ten compounds were isolated and identified as shancidin(1),coe-lonin(2),lusianthridin(3),orchinol(4),isoshancidin(5),blestriarene A(6),(E)-3-(4-hydroxyphenyl)acrylic acid(7),3-phenylpropanoic acid(8),ethyl 2-(4-hydroxyphenyl)acetate(9),ethyl 2-(4-ethylphenyl)acetate(10).Besides,compounds 7-10 were isolated from B.striata for the first time.Furthermore,compound 6 exerted an excellent bacterial inhibi-tory effect with a minimum inhibitory concentration of 10 μg/mL(positive control amoxicillin 0.8 μg/mL).The antibacterial mechanism results indicated that compound 6 disrupted the bacterial membrane structure,leading to alkaline phosphatase and DNA leakage,which resulted in bacterial death.Compound 6 may exert antibacterial activity by destroying cell wall and cell membrane.
Network pharmacology,molecular docking and animal experimental studies were used to investigate the mechanism of dihydromyricetin(DHM)in improving renal fibrosis in type 2 diabetic db/db mice.Initially,a 10-week DHM intervention was observed to ameliorate renal fibrosis in db/db mice.The chemical structure and targets of DHM were further obtained by TCMSP and PharmMapper,and the disease targets were retrieved by DisGeNET database.Venn analysis was performed on DHM targets and disease targets,and the intersection targets were uploaded to the String database to construct the PPI net-work.The'drug-target-disease'network was constructed by Cytoscape software.The GO enrichment analysis and KEGG en-richment analysis of the intersection target genes were performed by the David database.At the same time,the top 10 intersec-tion targets of PPI network were visualized,and the top 5 intersection targets were subjected to molecular docking with DHM using PDB database,Pymol software and AutoDock Tools software.The first core target AKT and related signaling pathways were verified by Western blot.Animal experiments showed that DHM intervention could reduce the body weight of db/db mice and improve the levels of blood glucose,creatinine,urea nitrogen and urine protein in db/db mice.HE,Masson and PAS stai-ning showed that renal fibrosis of db/db mice was alleviated.In addition,a total of 37 intersections of drugs and disease tar-gets were obtained by network pharmacology,300 GO-related items and 108 related pathways were obtained by enrichment a-nalysis.Molecular docking results showed that DHM could spontaneously bind to key targets.Western blot analysis results showed that DHM intervention could reduce the protein expression of Notch1,NICD,Hes1 and Hey1 in the kidney of db/db mice,up-regulate the level of PTEN protein,inhibit the phosphorylation of AKT,thus improve renal fibrosis.Taken together,DHM may alleviate renal fibrosis in db/db mice by regulating Notch/PTEN/AKT pathway.
A novel fibrinolytic enzyme was isolated and purified from the fruiting bodies of Lyophyllum decastes.The N-termi-nal amino acid sequence of the enzyme was determined by Edman degradation method.The fruiting bodies of Lyophyllum de-castes was dried,crushed and extracted with phosphate solution to obtain crude enzyme solution.The crude enzyme solution was isolated and purified by using ammonium sulfate precipitation,Octyl-Sepharose Fast Flow hydrophobic chromatography,SP-Sepharose high performance ion chromatography and Source 15PHE hydrophobic chromatography to obtain a single compo-nent with fibrinolytic activity.The specific activity of the enzyme was 4 105.78 U/mg,the purification fold was 206.09,and the recovery rate of the activity was 30.91％.Native-PAGE and SDS-PAGE showed that the fibrinolytic enzyme reached elec-trophoretic purity and the molecular weight was 30.9 kDa.The N-terminal sequences of the fibrinolytic enzyme were deter-mined by the Edman degradation method,and the sequences was Gly-Ala-Val-Thr-Gln-Cys-Asn-Ala-Pro-Trp-Gly-Leu.By NCBI database comparison,it was found that the fibrinolytic enzyme was a novel fibrinolytic enzyme.The research provides a new idea and method for the research and development of fibrinolytic enzyme.
This study aimed to explore the key components of total flavonoids from Lamiophlomis Herba(LH)against rheuma-toid arthritis(RA),which can elucidate the active components of LH against RA.In this work,the fingerprint of 13 batches of the total flavonoids from LH was established,including eight common peaks,of which three were identified.Based on the effect of 13 batches of total flavonoids from LH on the activity of fibroblast-like synoviocytes(FLS),the spectrum-effect rela-tionship was established for the anti-RA effect of total flavonoids from LH.The results showed that luteoloside and luteolin were key components of total flavonoids from LH against RA.Then,the component knockout technique was used to investigate the anti-RA efficacy of the luteoloside component in the total flavonoids from LH,and the effects of luteoloside and luteolin on FLS cell viability were compared.The results showed that luteoloside and luteolin could significantly inhibit FLS cell viability(P＜0.01),and there was no significant difference between their inhibition rates.Because the HPLC spectrum of total flavonoids form LH showed that the content of luteoloside was higher than that of luteolin,luteoloside is a key component of total flavonoids from LH against RA.This work can provide a certain reference for the clinical application of LH in the treatment of RA.
Tryptanthrin is a natural indolequinazoline alkaloid,is one of the bioactive components of many traditional Chinese herbal medicines,such as Indigo Naturalis,Isatidis Folium and Isatidis Radix.The antibacterial and anti-inflammatory effects of tryptanthrin have been reported for a long time.Recently,some studies have found that it also has anti-tumor,anti-virus,an-ti-angiogenesis,anti-parasitic and other pharmacological activities.Coupled with the characteristics of stable structure,low tox-ic and side effects,and easy absorption by the intestinal tract,it has become a current research hotspot.This review focused on the pharmacological activities of tryptanthrin,and at the same time summarized its natural sources,synthetic methods,as well as absorption,distribution,metabolism and toxicity in vivo.It is expected to provide a more comprehensive literature informa-tion support for the further research,development and utilization of tryptanthrin.
To investigate the mechanism and experimental verification of Morinda citrifolia in the treatment of atherosclerosis(AS)via network pharmacology.CMAUP,TCMSP and SwissADME databases were used to search and screen the active in-gredients of M.citrifolia.The AS targets were obtained after setting screening conditions in GeneCards,OMIM,TTD,PharmGKB and Drugbank databases.The protein interaction network(PPI)was constructed using STRING database,and the GO function enrichment analysis and KEGG pathway analysis for intersection targets were performed using R language.The THP-1-derived macrophage model was constructed for in vitro experiment verification.The concentration of M.citrifolia was screened by CCK-8 assay.Oil red O staining and 22-NBD-Cholesterol fluorescence assay were used to detect the intracellular cholesterol in the model cells following the treatment of M.citrifolia.A total of 59 active ingredients,332 targets and 154 tar-gets associated with AS,such as PPARG,MMP9,IL-6 and CCL2,were screened from M.citrifolia.GO function enrichment a-nalysis obtained 2 844 items,and the top 10 items mainly included biological functions such as regulating cell membrane re-ceptors and nuclear receptors.KEGG pathway analysis obtained 182 items,which of the top 20 items primarily contained lipid metabolism and atherosclerosis,peroxisome proliferator-activated receptor(PPAR)signaling pathway,etc.In vitro experi-ments were performed to validate the appropriate concentration of M.citrifolia(10,20,40 μg/mL)screened by CCK-8 assay firstly.Oil red O staining and 22-NBD-Cholesterol cell fluorescence found that M.citrifolia promoted cholesterol efflux of THP-1-derived macrophages in a dose-dependent manner.Subsequently,RT-PCR and WB assays showed that M.citrifolia ac-tivated PPARγ signaling pathway and increased the expression of PPARγ and ATP-binding cassette transporter A1(ABCA1)significantly.The capability of M.citrifolia to promote cholesterol efflux in THP-1-derived macrophages was decreased dramat-ically,and the levels of PPARγ and ABCA1 were down-regulated after blunting PPARγ signaling pathway by GW9662,PPARγ inhibitor.Therefore,network pharmacoloty combined with experimental validation in vitro indicated that M.citrifolia could ameliorate atherosclerosis by promoting cholesterol efflux from THP-1-derived macrophages via PPARγ signaling path-way.
This study explored the chemical components of fruits of the Chinese medicine Amomum tsao-ko through systematic separation.Various column chromatography techniques were used to separate and purify the ethanol extract and the structure of the compounds was identified mainly by spectral analysis.The inhibitory activity of the compounds on α-glucosidase was al-so studied by a PNPG method.The 11 compounds isolated from the ethyl acetate extract of the ethanol extract of the fruits of Amomum tsao-ko were named(R)-l-(1-ethoxypropyl)-3,5-dimethoxyphenol(1),(R)-l-(3,4,5-trimethoxyphenyl)propan-1-ol(2),4-hydroxy-3-methoxypropiophenone(3),apocynin(4),4-methyl-2,6-dimethoxyphenol(5),4-hydroxy-3-methoxy-benzaldehyde(6),1-terpinen-4-ol(7),methyl(9S,10R,11E,13R,15Z)-9,10,13-trihydroxyoctadeca-11,15-dienoate(8),methyl(9S,10R,11E,13R)-9,10,13-trihydroxyoctadec-11-enoate(9),amomutsaoko A(10)and renealtin A(11).Compound 1 is a phenolic compound and first reported as a natural product,compounds 10 and 11 have stronger inhibitory ac-tivity against α-glucosidase than acarbose.
To provide reference for the formulation of quality standards of Lobeliae Chinensis Herba formula granules and re-lated preparations,the transmission law of quantity value of Lobeliae Chinensis Herba standard decoction was studied.Twenty-one batches of Lobeliae Chinensis Herba decoction pieces were made into standard decoction.Paste yield and extracts were measured.The fingerprints of Lobeliae Chinensis Herba decoction pieces and standard decoction were established.Through similarity evaluation and chemometric analysis,the change rules of common peaks from decoction pieces to standard decoction were studied.The contents of diosmin and linarin were measured and the transfer rates were calculated.The results showed that paste yield of 21 batches of Lobeliae Chinensis Herba ranged from 29.91％to 47.94％and the extracts ranged from 22.19％-36.36％.Eleven identical common peaks were identified in the fingerprints of Lobeliae Chinensis Herba decoction pieces and standard decoction,indicating that the common peaks can be transferred from the decoction pieces to the standard decoction.The overall similarity of the fingerprint of the decoction pieces was relatively high,while the similarity of the stand-ard decoction varied greatly.Principal component analysis(PCA)showed that 21 batches of decoction pieces and standard decoction were classified into one group respectively.Five chromatographic peaks with significant changes in relative peak ar-ea were found by orthogonal partial least squares discriminant analysis(OPLS-DA).The content range of diosmin in 21 bat-ches of standard decoction was 0.15-0.54 mg/g and the transfer rate range was 1.53％-4.99％.The content range of linarin was 0.06-0.15 mg/g and the transfer rate range was 4.27％-8.98％.This study can provide a certain reference for the for-mulation of quality standards of Lobeliae Chinensis Herba formula granules and related preparations.
本实验探究白光(400～700 nm)、红光(620～650 nm)和绿光(500～580 nm)对等鞭金藻(Isochrysis sp.ISO-FJ)的生长和生物活性产物积累的协同影响,以期摸索适宜的光质同时提高其生物活性产物.实验结果表明,等鞭金藻细胞在白光条件下具有最大的生物量.在生物活性产物合成方面,与白光和红光相比,采用绿光培养的等鞭金藻细胞更有利于岩藻黄素、胞外多糖和多不饱和脂肪酸的联产合成.这些结果可表明,等鞭金藻的生长和活性产物合成是受不同光质的调节,为今后定向调控等鞭金藻生长和活性产物合成提供实践依据和理论指导.
探讨辣木叶多糖(Moringa oleifera leaf polysaccharide,MOLP)对葡聚糖硫酸钠(DSS)诱导的小鼠溃疡性结肠炎(ulcerative colitis,UC)防治作用及其作用机制.50只BALB/c小鼠随机分为正常对照组(Con组)、模型对照组(DSS 组)、MOLP-L(25 mg/kg)、MOLF-M(50 mg/kg)、MOLF-H(100 mg/kg)组,通过饮用 4％DSS 诱导小鼠 UC 模型.试验期间每日记录各组小鼠体重情况和疾病活动指数(DAI)变化;ELISA法检测血清中TNF-α、IL-1β、IL-10、HMGB1和结肠组织MPO含量;HE染色观察结肠组织病理学变化;实时荧光定量PCR法检测结肠组织TNF-α、IL-1β、IL-10、HMGB1 mRNA表达水平;蛋白免疫印迹(Western blot)法检测结肠组织中TLR4、MyD88、P65、p-P65、IκBα和p-IκBα的表达;16S rRNA高通量测序探究小鼠肠道菌群的变化.结果显示,与Con组相比,DSS组小鼠一般状况下降,体重减轻,结肠长度缩短,DAI和病理切片损伤评分均显著增加(P＜0.001);TNF-α、IL-1β、HMGB1含量和MPO活性显著升高(P＜0.001),结肠组织中TLR4、MyD88、p-P65和p-IκBα的蛋白表达水平显著上调(P＜0.001).与DSS组相比,不同剂量MOLP处理组明显改善上述病变和炎性指标,显著提升了 IL-10的表达,作用结果具有剂量依赖性;并且MOLP可以改善UC小鼠肠道菌群多样性,恢复菌群平衡.研究结果提示MOLP可以通过抑制炎症反应,调节小鼠肠道菌群多样性、组成和相对丰度进而发挥防治UC的作用.
