New insights into the interactions between the gut microbiota and the inflammatory response to ulcerative colitis in a mouse model of dextran sodium sulfate and possible mechanisms of action for treatment with PE&AFWE

Background:Inflammatory bowel disease(IBD),comprising Crohn's disease(CD)and ulcerative colitis(UC),is a heterogeneous state of chronic intestinal inflammation.Intestinal innate immunity,including innate immune cells,defends against pathogens and excessive entry of gut microbiota,while preserving immune tolerance to resident intestinal microbiota,and may be characterized by its capacity to produce a rapid and nonspecific reaction.The association between microbiota dysbiosis and the pathogenesis of IBD is complex and dynamic.When the intestinal ecosystem is in dysbiosis,the reduced abundance and diversity of intestinal gut microbiota make the host more vulnerable to the attack of exogenous and endogenous pathogenic gut microbiota.The aim of our study was to comprehensively assess the relationship between microbial populations within UC,the signaling pathways of pathogenic gut microbe therein and the inflammatory response,as well as to understand the effects of using PE&AFWE(poppy extract[Papaver nudicaule L.]and Artemisia frigida Willd.extract)on UC modulation.Methods:A UC mouse model was established by inducing SPF-grade C57BL/6 mice using dextrose sodium sulfate(DSS).Based on metagenomic sequencing to characterize the gut microbiome,the relationship between gut microbiota dysbiosis and gut microbiota was further studied using random forest and Bayesian network analysis methods,as well as histopathological analysis.Results:(1)We found that the 5 gut microbiota with the highest relative abundance of inflammatory bowel disease UC model gut microbiota were consistent with the top 5 ranked natural bacteria.There were three types of abundance changes in the model groups:increases(Chlamydiae/Proteobacteria and Deferribacteres),decreases(Firmicutes),and no significant changes(Bacteroidetes).The UC model group was significantly different from the control group,with 1308 differentially expressed species with abundance changes greater than or equal to 2-fold.(2)The proportion of the fecal flora in the UC group decreased by 37.5%in the Firmicutes and increased by 14.29%in the proportion of Proteobacteria compared to the control group before treatment.(3)The significantly enriched and increased signaling pathways screened were the'arachidonic acid metabolic pathway'and the'phagosomal pathway',which both showed a decreasing trend after drug administration.(4)Based on the causal relationship between different OTUs and the UC model/PE&AFWE administration,screening for directly relevant OTU networks,the UC group was found to directly affect OTU69,followed by a cascade of effects on OTU12,OTU121,OTU93,and OTU7,which may be the pathway of action that initiated the pathological changes in normal mice.(5)We identified a causal relationship between common differentially expressed OTUs and PE&AFWE and UC in the pre-and post-PE&AFWE-treated groups.Thereby,we learned that PE&AFWE can directly affect OTU90,after which it inhibits UC,inhibiting the activity of arachidonic acid metabolic pathway by affecting OTU118,which in turn inhibits the colonization of gut microbiota by OTU93 and OTU7.(6)Histopathological observation and scoring(HS)of the colon showed that there was a significant difference between the model group and the control group(p<0.001),and that there was a significant recovery in both the sulfasalazine(SASP)and the PE&AFWE groups after the administration of the drug(p<0.0001).Conclusion:We demonstrated causal effects and inflammatory metabolic pathways in gut microbiota dysbiosis and IBD,with five opportunistic pathogens directly contributing to IBD.PE&AFWE reduced the abundance of proteobacteria in the gut microbiota,and histopathology showed significant improvement.

Integrated mass spectrometry imaging reveals spatial-metabolic alteration in diabetic cardiomyopathy and the intervention effects of ferulic acid

Diabetic cardiomyopathy(DCM)is a metabolic disease and a leading cause of heart failure among people with diabetes.Mass spectrometry imaging(MSI)is a versatile technique capable of combining the molecular specificity of mass spectrometry(MS)with the spatial information of imaging.In this study,we used MSI to visualize metabolites in the rat heart with high spatial resolution and sensitivity.We optimized the air flow-assisted desorption electrospray ionization(AFADESI)-MSI platform to detect a wide range of metabolites,and then used matrix-assisted laser desorption ionization(MALDI)-MSI for increasing metabolic coverage and improving localization resolution.AFADESI-MSI detected 214 and 149 metabolites in positive and negative analyses of rat heart sections,respectively,while MALDI-MSI detected 61 metabolites in negative analysis.Our study revealed the heterogenous metabolic profile of the heart in a DCM model,with over 105 region-specific changes in the levels of a wide range of metabolite classes,including carbohydrates,amino acids,nucleotides,and their derivatives,fatty acids,glycerol phospholipids,carnitines,and metal ions.The repeated oral administration of ferulic acid during 20 weeks significantly improved most of the metabolic disorders in the DCM model.Our findings provide novel insights into the molecular mechanisms underlying DCM and the potential of ferulic acid as a therapeutic agent for treating this condition.

A review on cocrystal of active ingredients in traditional Chinese medicine

In the development of new drugs products,especially the development of traditional Chinese medicine active ingredients,solubility and oral bioavailability are the main factors which are restricting the development of new drugs,whereas the physicochemical properties of active ingredients are the key element to affecting these factors.Pharmaceutical cocrystal provides an excellent opportunity to develop new drugs with excellent physical and chemical properties such as melting point,solubility,stability and bioavailability while retaining the pharmacological properties of individuals active pharmaceutical ingredients among pharmaceutical cocrystal compounds.Traditional Chinese herbal medicine has the characteristics of multiple pathways and multiple targets,mainly because it contains many active ingredients,like cocrystals thereof with many components.The active ingredients extracted from traditional Chinese herbal medicine have a wide range of pharmacological activities,but most of the active ingredients affect the development of traditional Chinese medicine active ingredients due to their physical and chemical properties such as solubility.Traditional Chinese medicine pharmaceutical cocrystals can not only improve the physical and chemical properties of drugs without changing the internal structure of drugs,so as to provide a new scheme for the development of traditional Chinese medicine active ingredients.This paper reviews the research progress of active ingredients of traditional Chinese medicine pharmaceutical cocrystal.The preparation methods of cocrystals are summarized and the advantages of cocrystals are illustrated with examples.

Botanical,quality control,phytochemistry,pharmacology and toxicity characteristics of Corydalis bungeana Turcz.:a review

Corydalis bungeana Turcz.(CB)is a medicinal herb with significant medicinal value in traditional Chinese medicine.This paper reviews the progress of research on CB’s botany,quality control,phytochemistry,pharmacology and toxicity.The plant’s information was gathered from scientific databases such as PubMed,GeenMedical,Springer Link(https://link.springer.com),Chinese National Knowledge Infrastructure,Pharmacopoeia and Flora.Currently,137 phytochemicals have been identified and isolated from CB,including alkaloids,flavonoids,amino acids,terpenoids,coumarins and organic acids.In addition,many phytochemicals reported various antiinflammatory,antibacterial,antiviral,antitumor,analgesic,hepatoprotective,immunomodulatory,neuromodulatory,and lipid reduction activities.However,the study of its toxicity is still at the preliminary exploration stage and needs further intensive exploration.Herein,we provide an in-depth investigation of the progress of CB to elucidate the underlying mechanisms of activity of CB extracts and its major components,deliver valuable resources and information for further research and rational drug use,and explore the potential research directions and prospects of CB.

Plant distribution and pharmacological activity of flavonoids

Flavonoids are natural organic compounds that are widely found in nature, their structural types are complex, and they mainly include flavonoids, flavonols, dihydroflavonols, isoflavones, dihydroisoflavones, chalcones, orange ketones, flavanoids, anthocyanidins, and biflavonoids. This review covers the plant distribution and pharmacological activities of flavonoids. Flavonoids are mainly distributed in angiosperms and gymnosperms, and they are abundant in plants such as Rutaceae, Labiatae, Zingiberaceae, Scrophulariaceae, and Leguminosae. Because of their wide distribution and variety, researchers have found that flavonoids have diverse biological activities, mainly focusing on anti-inflammatory, antibacterial and antitumor activities. Mechanistically, the anti-inflammatory effects are mainly related to the NF-κB and MAPK (mitogen-activated protein kinase) signaling pathway and then the inhibition of the production of inflammatory cytokines and mediators. The antibacterial activity is mainly manifested as inhibitory effects on many strains, including Escherichia coli, Cryptococcus neoformans, and Pseudomonas aeruginosa, via destroying the stability of the microbial membrane, inhibiting the invasion of virulent bacteria into host cells, promoting the apoptosis of bacteria, inhibiting bacterial fatty acid synthesis, etc. The antitumor activity of flavonoids is related to their inhibition of cell proliferation and induction of apoptosis via the mitochondria-mediated, endoplasmic reticulum-mediated, and death factor and its receptor-mediated signal transduction pathways. Understanding the plant distribution and pharmacological activity of flavonoids not only reveals the importance of identifying such valuable flavonoids in another genus or family but also provides a basis for fully exploiting the therapeutic potential of flavonoids.

Ethnobotany and diversity of medicinal plants used by the Buyi in eastern Yunnan, China

The Buyi are a socio-linguistic group in Yunnan Province of southwest China that have a long history of using medicinal plants as part of their indigenous medical system.Given the limited written documentation of the Buyi indigenous medical system,the objective of this paper is to document the medicinal plants of the Buyi and associated traditional knowledge and transmission.Field research was conducted in four villages in Lubuge Township of Luoping County in Yunnan Province using ethnobotanical methodologies including participatory observation,semi-structured interviews,key informant interviews,and focus group discussions to elicit information on medicinal plants.In total,120 informants(including 15 key informants who are healers)were interviewed.This study found that a total of 121 medicinal plant species belonging to 64 families are used by the Buyi including by local healers to treat different diseases.Among the medicinal plants recorded in this study,56 species(46%)have not previously been documented in the scientific literature as having medicinal value,highlighting the pressing need for ethnobotanical documentation in indigenous communities.The most frequently used medicinal part was the leaf(24.9%of documented plants),and the most common preparation method was decoction(62.8%of medicinal).Medicinal plants were mainly used to treat rheumatism(12.4%of plants),trauma and injuries(9.6%).The documented plants are also used for other non-medicinal purposes including food,fodder,fencing,and ornamental.In addition,35 of the medicinal plants are considered poisonous and are used by local Buyi healers for medicine.The traditional Buyi beliefs and practices associated with the documented medicinal plants likely contributes to their conservation in the environments and around Buyi communities.This study further highlights that ethnomedicinal knowledge of the Buyi is at risk of disappearing due to increased introduction and use of modern medicine in Buyi communities,livelihood changes,rapid modernization,and urbanization.Research,policy,and community programs are urgently needed to conserve the biocultural diversity associated with the Buyi medical system including ethnobotanical knowledge towards supporting both environmental and human wellbeing.

Folk nomenclature of plants in Cistanche deserticola-associated community in South Gobi, Mongolia

Cistanche deserticola is an important medicinal plant in Mongolia.Despite its significant role in local healing systems,little traditional knowledge had been reported.The present study investigated folk names of C.deserticola and other species of the same community in Umnugobi Province,South Gobi region of Mongolia,based on ethnobotanical approaches.The high correspondence between folk names and scientific names of plant species occurring in Cistanche-associated community shows the scientific meaning of folk nomenclature and classification in Mongolia.The Mongolian and folk names of plants were formed on the basis of observations and understanding of wild plants including their morphology,phenology and traditional uses as well.Results from this study will support the conservation of C.deserticola itself,a rare and endangered plant species listed in the Monglian Red Data Book.Our documentation of folk nomenclature based on 96 plant species in the Cistanche community,as a part of traditional knowledge associated with biodiversity,will be very helpful for making strategy of plant biodiversity conservation in Mongolia.

Identification of potential anti-pneumonia pharmacological components of Glycyrrhizae Radix et Rhizoma after the treatment with Gan An He Ji oral liquid

Glycyrrhizae Radix et Rhizoma,a traditional Chinese medicine also known as Gan Cao(GC),is frequently included in clinical prescriptions for the treatment of pneumonia.However,the pharmacological components of GC for pneumonia treatment are rarely explored.Gan An He Ji oral liquid(GAHJ)has a simple composition and contains GC liquid extracts and paregoric,and has been used clinically for many years.Therefore,GAHJ was selected as a compound preparation for the study of GC in the treatment of pneumonia.We conducted an in vivo study of patients with pneumonia undergoing GAHJ treatments for three days.Using the intelligent mass spectrometry data-processing technologies to analyze the metabolism of GC in vivo,we obtained 168 related components of GC in humans,consisting of 24 prototype components and 144 metabolites,with 135 compounds screened in plasma and 82 in urine.After analysis of the metabolic transformation relationship and relative exposure,six components(liquiritin,liquiritigenin,glycyrrhizin,glycyrrhetinic acid,daidzin,and formononetin)were selected as potential effective components.The experimental results based on two animal pneumonia models and the inflammatory cell model showed that the mixture of these six components was effective in the treatment of pneumonia and lung injury and could effectively downregulate the level of inducible nitric oxide synthase(iNOS).Interestingly,glycyrrhetinic acid exhibited the strongest inhibition on iNOS and the highest exposure in vivo.The following molecular dynamic simulations indicated a strong bond between glycyrrhetinic acid and iNOS.Thus,the current study provides a pharmaceutical basis for GC and reveals the possible corresponding mechanisms in pneumonia treatment.

Amides,Isoquinoline Alkaloids and Dipeptides from the Aerial Parts of Piper mullesua

One undescribedamide,pipermullesine A,twoundescribed isoquinoline alkaloids,pipermullesinesBand C,and six undescribed dipeptides,pipermullamides A–F,along with 28 known compounds,were isolated from the aerial parts of Piper mullesua.The structures of the undescribed compounds were elucidated based on the analysis of 1D and 2D NMR and MS data.Furthermore,the structures of pipermullesines A–Cwere confirmed by single crystal X-ray diffraction analysis.All isolates were evaluated for inhibitory activity against platelet aggregation induced by thrombin(IIa)or platelet-activating factor(PAF).(-)-Mangochinine,pellitorine,and(2E,4E)-N-isobutyl-2,4-dodecadienamide showed weak inhibitory activity against rabbit platelet aggregation induced by PAF,with IC_(50)values of 470.3μg/mL,614.9μg/mL,and 579.7μg/mL,respectively.

YINDARA-4 relieves visceral hypersensitivity in irritable bowel syndrome rats via regulation of gut microbiota and serotonin levels

Objective:The present study aims to evaluate the in vivo efficacy of YINDARA-4 in improving the symptoms of irritable bowel syndrome(IBS)in a rat model and investigate the impact of YINDARA-4 on potential targets of IBS management,such as the serotonin level in intestinal tissues and the structure and composition of the gut microbiota.Methods:We developed an IBS rat model by combining stress from maternal separation,acetic acid administration,and restraint.We administered YINDARA-4 water extract to the IBS rat model for 10 consecutive days.The fecal water content,visceral sensitivity,gut microbiota,and serotonin levels in the colonic tissue were then analyzed and compared between the control group,IBS model group,and YINDARA-4–treated groups.Results:Treatment with YINDARA-4 reversed visceral hypersensitivity in a dose-dependent manner in the experimental rat model of IBS.The relief of visceral hypersensitivity upon treatment with YINDARA-4 involved regulation of the gut microbiota structure and composition,and normalization of elevated serotonin levels in the colon.The decrease in colonic serotonin levels with YINDARA-4 treatment might be associated with a reduction in the abundance of Helicobacter and enrichment of Butyricimonas.Conclusions:Treatment with YINDARA-4 was beneficial against visceral hypersensitivity in a rat model of IBS.The improved symptoms exhibited in IBS rats were associated with favorably altered gut microbiota and normalization of serotonin levels in the colon.

Research progress on anti-tumor properties of Marsdenia tenacissima

Tongguanteng （Marsdenia tenacissima）, which is mainly distributed in the Yunnan and Guizhou provinces of China, wasfirst recorded in Diannanbencao by Lan Mao of the Ming dynasty of China. According to recent pharmacological studies,the chemical composition of Tongguanteng （Marsdenia tenacissima） is complex and contains C21 steroidal saponins,polysaccharides, alkaloids, and other molecules, which show anti-cancer effects on various tumor cell lines. It inhibitstumor cell proliferation and growth mainly by increasing the expression of apoptosis- and cell cycle-related proteins topromote apoptosis and arrest tumor cells in the G2/M or S phase. Downregulation of the expression of vascularendothelial growth factor-2/A and matrix metalloprotease-2/9 suppresses the formation of the tumor microvasculature,leading to tumor malnutrition, increased expression of interleukin-2, glutathione peroxidase, catalase, and superoxidedismutase, and decreased interleukin-10 and malondialdehyde expression, thereby enhancing immunity andantioxidation in the body. Additionally, inhibition of epidermal growth factor receptor, hepatocyte growth factor receptor,and tyrosine-protein kinase receptor activation enhances the anti-tumor efficacy of epidermal growth factorreceptor-tyrosine kinase inhibitors as well as inhibits P-glycoprotein and cytochrome P450 to increase the concentrationof anti-tumor drugs in tumor cells.

Effect of Zhuang Medicine Feilongzhangxue on the Expression of HMGB1-TLR4 / RANKL-NF-κB Signaling Pathway Related Factors in Osteoclasts

Objective:To study the regulatory effect of feilongzhangxue on the levels of HMGB1-TLR4/RANKL-NF-κB signaling pathway related factors HMGB1,RANKL,rank,TRAF-6 and NF-κBp65 in osteoclasts,so as to explore the mechanism of feilongzhangxue intervention in RA;Methods:The osteoclasts with good activity were randomly divided into blank group,methotrexate control group and Zhuang medicine feilongzhang blood containing serum treatment group,which were divided into OC+blank group,OC+methotrexate control group,OC+Zhuang medicine feilongzhang blood containing serum group;The expression of HMGB1,RANKL,rank,TRAF-6 and NF-κBp65 mRNA was detected by RT-PCR;The protein expressions of HMGB1,RANKL,RANK,TRAF-6 and NF-κBp65 were detected by immunofluorescence.Results:PCR results showed that:Compared with the blank group,feilongzhangxue could effectively inhibit the expression levels of HMGB1,RANKL,rank,TRAF-6 and NF-κB p65 mRNA in OC cells,and the inhibitory effect was stronger than methotrexate.Immunofluorescence test results showed that:Compared with the blank group and methotrexate group,feilongzhangxue could effectively inhibit the protein expression of HMGB1,RANKL,rank,TRAF-6 and NF-κB p65 in OC cells.Conclusion:The effect of Zhuang medicine feilongzhangxue on hmgbl-tlr4/rankl-nf-κB signaling pathway of osteoclasts is through the regulation of related factors HMGB1,RANKL,rank,TRAF-6 and NF-κB p65,which may be the key mechanism of Zhuang medicine feilongzhangxue on rheumatoid arthritis.

Chemical profiling of principle active and toxic constituents in herbs containing aristolochic acids

Objective:To clear the amounts of the principal active/toxic components in herbs containing aristolochic acids(HCAAs),which are still used as medicine and/or seasoning in many ethnic minority areas of China.Methods:In this study,six major active and toxic components in HCAAs were extracted with ultrasonic extraction.With 6-O-methyl guanosine as internal standard,the target compounds were analyzed qualitatively and quantitatively by using ultrahigh performance liquid chromatography-electrospray ionization-tandem mass spectrometry(UPLC-ESI-MS/MS)with multiple reaction monitoringinformation dependent acquisition-enhanced production ion scanning mode(MRM-IDA-EPI)combined with dynamic background subtraction(DBS)function.Results:The method showed good linearity in the linear range of the six analytes.The limit range of detection was from 0.01 ng/mL to 0.27 ng/mL.All of the detection repeatability,extraction repeatability and accuracy of the method were good.After extraction,the samples remained stable at 15℃ within 24 h.Six analytes were all found in samples except aristolactam(AL)in sample 2,and the contents varied greatly.The contents of these compounds decreased in fruits,leaves and stems of Aristolochia delavayi successively.Conclusion:This method has the advantages of less sample dosage,simple operation,short analysis cycle,high sensitivity,specificity and accuracy.It laid a good foundation for guiding the safety of HCAAs,the indepth study of pharmacological and toxicological effects and the scientific and standardized processing and compatibility of HCAAs.

Research progress on the relationship between intestinal microecology and intestinal bowel disease

Intestinal microecology is the main component of human microecology.Intestinal microecology consists of intestinal microbiota,intestinal epithelial cells,and intestinal mucosal immune system.These components are interdependent and establish a complex interaction network that restricts each other.According to the impact on the human body,there are three categories of symbiotic bacteria,opportunistic pathogens,and pathogenic bacteria.The intestinal microecology participates in digestion and absorption,and material metabolism,and inhibits the growth of pathogenic microorganisms.It also acts as the body’s natural immune barrier,regulates the innate immunity of the intestine,controls the mucosal barrier function,and also participates in the intestinal epithelial cells’physiological activities such as hyperplasia or apoptosis.When the steady-state balance of the intestinal microecology is disturbed,the existing core intestinal microbiota network changes and leads to obesity,diabetes,and many other diseases,especially irritable bowel syndrome,inflammatory bowel disease(IBD),and colorectal malignancy.Intestinal diseases,including tumors,are particularly closely related to intestinal microecology.This article systematically discusses the research progress on the relationship between IBD and intestinal microecology from the pathogenesis,treatment methods of IBD,and the changes in intestinal microbiota.

Directional screening and identification of potential cytotoxic components from Achnatherum inebrians by a combination of surface palsmon resonance and chromatography

Objective:To establish a method for directional screening of the cytotoxic components from the medicinal herb of Achnatherum inebrians by a combination of surface plasmon resonance(SPR)biosensor and chromatographic isolation technology.Methods:Under the guidance of bioactive assessment based on binding abilities between objects and the a-Mannosidase(a-Man)target,the active components from different solvents extracts,different polar extraction parts and fractions were screened orderly and directionally using SPR.Components with a high binding ability to a-Man can be precisely oriented in a narrower fractions range and are easy to isolate.Three human cancer cells were used to evaluate the cytotoxic activity of component with the highest affinity to a-Man.Results:Eight compounds were isolated and identificated from A.inebrians for the first time.Deoxyvasicinone possessed the highest affinity to a-Man among them.Moreover,deoxyvasicinone showed good effects on inhibited proliferation of human hepatoma cells HepG2(IC_(50)=5.7 μmol/L),human breast cancer cells MCF7(IC_(50)=7.21 μmol/L)and human lung cancer cells HCC827(IC_(50)=0.75 μmol/L),respectively.In particular,its inhibitory effect on HCC827 was stronger than the positive drug gefitinib(IC_(50)=1.65 μmol/L).Conclusion:A comprehensive strategy of directional screening potential cytotoxic components from herb based on biomolecular interaction and chromatography was established.Deoxyvasicinone as an effective anti-cancer component was initially isolated from A.inebrians.It is expected that this screening strategy could provide new perspectives for rapid screening and identification of active components from natural plants with the complex matrix.

Comprehensive chemical profiling and quantitative analysis of ethnic Yi medicine Miao-Fu-Zhi-Tong granules using UHPLC-MS/MS

Developing analytical methods for the chemical components of natural medicines remains a challenge due to its diversity and complexity.Miao-Fu-Zhi-Tong(MFZT)granules,an ethnic Yi herbal prescription,comprises 10 herbs and has been clinically applied for gouty arthritis(GA)therapy.Herein,a series of chemical profiling strategies including in-house library matching,molecular networking and MS/MS fragmentation behavior validation based on ultra-high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS)were developed for qualitative analysis of MFZT granules.A total of 207 compounds were identified or characterized in which several rare guanidines were discovered and profiled into alkyl substituted or cyclic subtypes.Moreover,network pharmacology analysis indicated that MFZT’s anti-gout mechanism was mostly associated with the nuclear factor kappa-B(NF-κB)signaling,nucleotide oligomerization domain(NOD)-like signaling and rheumatoid arthritis pathways,along with the synergistic effect of 84 potential active compounds.In addition,a quantitative analytical method was developed to simultaneously determine the 29 potential effective components.Among them,berberine,pellodendrine,3-feruloylquinic acid,neoastilbin,isoacteoside and chlorogenic acid derivatives at higher concentrations were considered as the chemical markers for quality control.These findings provide a holistic chemical basis for MFZT granules and will support the development of effective analytical methods for the herbal formulas of natural medicines.

In situ metabolomics in nephrotoxicity of aristolochic acids based on air flow-assisted desorption electrospray ionization mass spectrometry imaging

Understanding of the nephrotoxicity induced by drug candidates is vital to drug discovery and development.Herein,an in situ metabolomics method based on air flow-assisted desorption electrospray ionization mass spectrometry imaging(AFADESI-MSI)was established for direct analysis of metabolites in renal tissue sections.This method was subsequently applied to investigate spatially resolved metabolic profile changes in rat kidney after the administration of aristolochic acid I,a known nephrotoxic drug,aimed to discover metabolites associated with nephrotoxicity.As a result,38 metabolites related to the arginine-creatinine metabolic pathway,the urea cycle,the serine synthesis pathway,metabolism of lipids,choline,histamine,lysine,and adenosine triphosphate were significantly changed in the group treated with aristolochic acid I.These metabolites exhibited a unique distribution in rat kidney and a good spatial match with histopathological renal lesions.This study provides new insights into the mechanisms underlying aristolochic acids nephrotoxicity and demonstrates that AFADESI-MSI-based in situ metabolomics is a promising technique for investigation of the molecular mechanism of drug toxicity.

Hypoglycemic effects of Tibetan medicine Huidouba in STZ-induced diabetic mice and db/db mice

Objective:Huidouba(HDB) is a Chinese folk medicine used to treat diabetes in Sichuan Province,China.Therefore,we investigated the anti-diabetic effects of HDB and its underlying mechanisms.We hypothesized that HDB treatment could enhance glucose tolerance and insulin sensitivity,and thus prevent a hyperglycemia state.Methods:To test the hypothesis,streptozotocin(STZ)-induced diabetic mice and db/db mice,widely used models of hyperglycemia and insulin-resistant diabetes,were either treated with HDB,metformin,or acarbose.Blood glucose,oral glucose tolerance test,insulin tolerance test,pancreatic histopathology and serum biochemistry were detected to assess the hypoglycemic effect of HDB.Results:HDB treatments were found to show the effect in reducing glucose levels.HDB also resulted in a significant reduction in body weight and food intake in the STZ-induced diabetic mouse model.Furthermore,it significantly improved glucose and insulin tolerance in the two diabetic mouse models.Importantly,insulin,glucagon,pancreatic polypeptide,and somatostatin immunohistochemistry revealed that HDB treatment improved the function and the location of the cells in the islets compared with the other two treatments.HDB treatment resulted in significant restoration of islet function.Our results illustrated the underlying mechanism of HDB in the progression of diabetes,and HDB can be an effective agent for the treatment of diabetes.Conclusion:The results of this study suggested that HDB can reduce blood glucose levels in STZ-induced hyperglycemic mice and db/db mice.

Functional and binding studies of gallic acid showing platelet aggregation inhibitory effect as a thrombin inhibitor

Objective:This study was devoted to identifying natural thrombin inhibitors from traditional Chinese medicine(TCM)and evaluating its biological activity in vitro and binding characteristics.Methods:A combination strategy containing molecular docking,thrombin inhibition assay,surface plasmon resonance(SPR)and molecular dynamics simulation were applied to verify the study result.Results:Gallic acid was confirmed as a direct thrombin inhibitor with IC;of 9.07μmol/L and showed a significant inhibitory effect on thrombin induced platelet aggregation.SPR-based binding studies demonstrated that gallic acid interacted with thrombin with a KDvalue of 8.29μmol/L.Molecular dynamics and binding free energy analysis revealed that thrombin-gallic acid system attained equilibrium rapidly with very low fluctuations,the calculated binding free energies was-14.61 kcal/mol.Ala230,Glu232,Ser235,Gly258 and Gly260 were the main amino acid residues responsible for thrombin inhibition by gallic acid,providing a mechanistic basis for further optimization.Conclusion:This study proved that gallic acid is a direct thrombin inhibitor with platelet aggregation inhibitory effect,which could provide a basis for the follow-up research and development for novel thrombin inhibitors.

Portulaca oleracea alleviates CCl4-induced acute liver injury by regulating hepatic S100A8 and S100A9

Objective: Acute liver injury(ALF) is a potential factor of many serious hepatopathies. Carbon tetrachloride(CCl4) is a possible environmental toxicant that can induce ALF. Portulaca oleracea(PO) is one of the most popular edible herbs and has several biological activities such as antioxidant, antimicrobial, antiinflammatory effects. We explored the significance of PO in regulating inflammatory function in animal models and cultured hepatocytes during liver damage caused by CCl4.Methods: The effect of PO on ALF was evaluated by CCl4-induced mice models in vivo. Hepatic levels of transaminase activities and inflammatory factors were examined. The gene and protein expression of S100A8 and S100A9 were measured by RT-PCR and Western blot analysis. Meanwhile, the efficacy of PO was certified by HepG2 cells in vitro. The transaminase activities, inflammatory factors, and the protein expression of S100A8 and S100A9 were also detected.Results: Animal tests showed that pretreatment with PO reduced the liver pathological tissue damage and the serum levels of ALT, AST, ALT and LDH, as well as reducing the pro-inflammatory cytokines(IL-1β, IL-6, TNF-a) secretion in CCl4-induced liver injury mice. Simultaneously, Hep G2 cells pretreated with PO exhibited a significant decrease in the activities of ALT and AST. Moreover, PO resulted in a significant downregulation of the pro-inflammatory markers S100A8, S100A9 gene and protein expression on CCl4induced acute liver injury was demonstrated entirely in vivo and vitro experiments.Conclusion: PO may down-regulate S100A8 and S100A9 and inhibit pro-inflammatory cytokines’ release,indicating a potential clinical effect for controlling the disease.