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Single-cell transcriptomic dissection of the cellular and molecular events underlying the triclosan-induced liver fibrosis in mice

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Military Medical research 2023年第5期10卷 599-619页

作者： Yun-Meng Bai Fan Yang Piao Luo Lu-Lin Xie Jun-Hui Chen Yu-Dong Guan Hong-Chao Zhou Teng-Fei Xu Hui-Wen Hao Bing Chen Jia-Hui Zhao Cai-Ling Liang Ling-Yun Dai Qing-Shan Geng Ji-Gang Wang Department of Nephrology Shenzhen Key Laboratory of Kidney Diseasesand Shenzhen Clinical Research Centre for GeriatricsShenzhen People’s Hospitalthe First Affiliated HospitalSouthern University of Science and TechnologyShenzhen 518020China Department of Urology Shenzhen People’s Hospitalthe First Affiliated HospitalSouthern University Science and Technologythe Second Clinical Medical CollegeJinan UniversityShenzhen 518020China integrated chinese and western medicine postdoctoral research station Jinan UniversityGuangzhou 510632China Artemisinin research Center and Institute of Chinese Materia MedicaChina Academy of Chinese Medical SciencesBeijing 100700China Guangdong Provincial Key Laboratory of New Drug Screening School of Pharmaceutical SciencesSouthern Medical UniversityGuangzhou 510515China Center for Reproductive medicine Dongguan Maternal and Child Health Care HospitalSouthern Medical UniversityDongguan 523125GuangdongChina

Background: Triclosan [5-chloro-2-(2,4-dichlorophenoxy) phenol, TCS], a common antimicrobial additive in many personal care and health care products, is frequently detected in human blood and urine. Therefore, it has been considered an emerging and potentially toxic pollutant in recent years. Long-term exposure to TCS has been suggested to exert endocrine disruption effects, and promote liver fibrogenesis and tumorigenesis. This study was aimed at clarifying the underlying cellular and molecular mechanisms of hepatotoxicity effect of TCS at the initiation stage.Methods: C57BL/6 mice were exposed to different dosages of TCS for 2 weeks and the organ toxicity was evaluated by various measurements including complete blood count, histological analysis and TCS quantification. Single cell RNA sequencing(scRNA-seq) was then carried out on TCS-or mock-treated mice livers to delineate the TCS-induced hepatotoxicity. The acquired single-cell transcriptomic data were analyzed from different aspects including differential gene expression, transcription factor(TF) regulatory network, pseudotime trajectory, and cellular communication, to systematically dissect the cellular and molecular events after TCS exposure. To verify the TCS-induced liver fibrosis,the expression levels of key fibrogenic proteins were examined by western blotting, immunofluorescence, Masson’s trichrome and Sirius red stainings. In addition, normal hepatocyte cell MIHA and hepatic stellate cell LX-2 were used as in vitro cell models to experimentally validate the effects of TCS by immunological, proteomic and metabolomic technologies.Results: We established a relatively short term TCS exposure murine model and found the TCS mainly accumulated in the liver. The scRNA-seq performed on the livers of the TCS-treated and control groups profiled the gene expressions of > 76,000 cells belonging to 13 major cell types. Among these types, hepatocytes and hepatic stellate cells(HSCs)were significantly increased in TCS-treated group. We found that TCS promoted fibros

关键词： Triclosan Single cell RNA sequencing Liver fibrogenesis Hepatic stellate cell

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Effects of Total Saponins from Dioscorea Nipponica Makino on Monosodium Urate-Induced M1-Polarized Macrophages through Arachidonic Acid Signaling Pathway:An in vitro Study

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chinese Journal of Integrative medicine 2023年第1期29卷 44-51页

作者： ZHOU qi SUN Hui-juan LIU Shu-min research Institute of chinese medicine Heilongjiang University of Chinese MedicineHarbin150040China Graduate School Heilongjiang University of Chinese MedicineHarbin150040China Technological Innovation Team of Basic Theory Study research of Institution of Higher Education in Heilongjiang Province Heilongjiang University of Chinese MedicineHarbin150040China

Objective To investigate and reveal the underlying mechanism of the effect of total saponins from Dioscoreae nipponica Makino(TSDN)on the arachidonic acid pathway in monosodium urate(MSU)crystal-induced M1-polarized macrophages.Methods M1 polarization of RAW264.7 cells were induced by 1µg/mL lipopolysaccharide(LPS).The methylthiazolyldiphenyl-tetrazolium bromide method was then used to screen the concentration of TSDN.MSU(500µg/mL)was used to induce the gouty arthritis model.Afterwards,10µg/L TSDN and 8µmol/L celecoxib,which was used as a positive control,were added to the above LPS and MSU-induced cells for 24 h.The mRNA and protein expressions of cyclooxygenase(COX)2,5-lipoxygenase(5-LOX),microsomal prostaglandin E synthase derived eicosanoids(mPGES)-1,leukotriene B(LTB)4,cytochrome P450(CYP)4A,and prostaglandin E_(2)(PGE_(2))were tested by real-time polymerase chain reaction and western blotting,respectively.The enzyme-linked immunosorbent assay was used to test the contents of M1 markers,including inducible nitric oxid synthase(NOS)2,CD80,and CD86.Results TSDN inhibited the proliferation of M1 macrophages and decreased both the mRNA and protein expressions of COX2,5-LOX,CYP4A,LTB4,and PGE_(2)(P<0.01)while increased the mRNA and protein expression of mPGES-1(P<0.05 or P<0.01).TSDN could also significantly decrease the contents of NOS2,CD80,and CD86(P<0.01).Conclusion TSDN has an anti-inflammation effect on gouty arthritis in an in vitro model by regulating arachidonic acid signaling pathway.

关键词： gouty arthritis Dioscorea nipponica Makino arachidonic acid signaling pathway M1/M2 polarization

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integrated'omics analysis for the gut microbiota response to moxibustion in a rat model of chronic fatigue syndrome

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Journal of Traditional chinese medicine 2023年第6期43卷 1176-1189页

作者： LI Chaoran YANG Yan FENG Chuwen LI Heng QU Yuanyuan WANG Yulin WANG Delong WANG Qingyong GUO Jing SHI Tianyu SUN Xiaowei WANG Xue HOU Yunlong SUN Zhongren YANG Tiansong Department of Acupuncture Zhejiang Chinese Medical UniversityHangzhou 310000China Department of chinese Medical Literature College of Basic MedicineHeilongjiang University of Chinese medicineHarbin 150040China Department of Rehabilitation the First Affiliated Hospital of Heilongjiang University of Traditional Chinese MedicineHarbin 150040China Department of Acupuncture the First Affiliated Hospital of Heilongjiang University of Traditional Chinese MedicineHarbin 150040China Graduate School the Second Affiliated Hospital of Heilongjiang University of Traditional Chinese MedicineHarbin 150040China Department of Acupuncture the Second Affiliated Hospital of Heilongjiang University of Traditional Chinese MedicineHarbin 150040China Shanghai Applied Protein Technology Co. Ltd.Shanghai 200233China Department of Acupuncture Chongqing Changshou District People's HospitalChongqing 401220China College of integrated chinese and western medicine Hebei University of Chinese Medicineand National Key Laboratory of Collateral Disease Research and Innovative Chinese MedicineHebei 050000China Department of Rehabilitation the First Affiliated Hospital of Heilongjiang University of Traditional Chinese Medicineand Traditional Chinese Medicine Informatics Key Laboratory of Heilongjiang ProvinceHarbin 150040China

OBJECTIVE:To observe the efficacy of moxibustion in the treatment of chronic fatigue syndrome(CFS)and explore the effects on gut microbiota and metabolic profiles.METHODS:Forty-eight male Sprague-Dawley rats were randomly assigned to control group(Con),CFS model group(Mod,established by multiple chronic stress for 35 d),MoxA group(CFS model with moxibustion Shenque(CV8)and Guanyuan(CV4),10 min/d,28 d)and MoxB group(CFS model with moxibustion Zusanli(ST36),10 min/d,28 d).Open-field test(OFT)and Morris-water-maze test(MWMT)were determined for assessment the CFS model and the therapeutic effects of moxibustion.16S rRNA gene sequencing analysis based gut microbiota integrated untargeted liquid chromatograph-mass spectrometer(LC-MS)based fecal metabolomics were executed,as well as Spearman correlation analysis,was utilized to uncover the functional relevance between the potential metabolites and gut microbiota.RESULTS:The results of our behavioral tests showed that moxibustion improved the performance of CFS rats in the OFT and the MWMT.Microbiome profiling analysis revealed that the gut microbiomes of CFS rats were less diverse with altered composition,including increases in pro-inflammatory species(such as Proteobacteria)and decreases in anti-inflammatory species(such as Bacteroides,Lactobacillus,Ruminococcus,and Prevotella).Moxibustion partially normalized these changes in the gut microbiota.Furthermore,CFS was associated with metabolic disorders,which were effectively ameliorated by moxibustion.This was demonstrated by the normalization of 33 microbiota-related metabolites,including mannose(P=0.001),aspartic acid(P=0.009),alanine(P=0.007),serine(P=0.000),threonine(P=0.027),methionine(P=0.023),5-hydroxytryptamine(P=0.008),alpha-linolenic acid(P=0.003),eicosapentaenoic acid(P=0.006),hypoxanthine(P=0.000),vitamin B6(P=0.000),cholic acid(P=0.013),and taurocholate(P=0.002).Correlation analysis showed a significant association between the perturbed fecal microbiota and metabolite levels,with a notable negative relations

关键词： moxibustion fatigue syndrome chronic gastrointestinal microbiome 16S rRNA gene sequencing metabolomics

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Biotransformation ofα-asarone by Alternaria longipes CGMCC 3.2875

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chinese Journal of Natural medicines 2021年第9期19卷 700-705页

作者： ZOU Jian ZHANG Shuai ZHAO Huan WANG Yong-Heng ZHOU Zheng-Qun CHEN Guo-Dong HU Dan LI Ning YAO Xin-Sheng GAO Hao Institute of Traditional chinese medicine&Natural Products College of Pharmacy/Guangdong Province Key Laboratory of Pharmacodynamic Constituents ofTCM and New Drugs ResearchJinan UniversityGuangzhou 510632China College of Traditional chinese Materia Medica Shenyang Pharmaceutical UniversityShenyang 110016China College of Traditional chinese medicine Jinan UniversityGuangzhou 510632China integrated chinese and western medicine postdoctoral research station Jinan UniversityGuangzhou 510632China

Biotransformation ofα-asarone by Alternaria longipes CGMCC 3.2875 yielded two pairs of new neolignans,(+)(7 S,8 S,7’S,8’R)iso-magnosalicin(1 a)/(-)(7 R,8 R,7’R,8’S)iso-magnosalicin(1 b)and(+)(7 R,8 R,7’S,8’R)magnosalicin(2 a)/(-)(7 S,8 S,7’R,8’S)magnosalicin(2 b),and four known metabolites,(±)acoraminol A(3),(±)acoraminol B(4),asaraldehyde(5),and 2,4,5-trimethoxybenzoic acid(6).Their structures,including absolute configurations,were determined by extensive analysis of NMR spectra,X-ray crystallography,and quantum chemical ECD calculations.The cytotoxic activity and Aβ_(42)aggregation inhibitory activity of all the compounds were evaluated.Compound 2 displayed significant anti-Aβ_(42)aggregation activity with an inhibitory rate of 60.81%(the positive control EGCG:69.17%).In addition the biotransformation pathway ofα-asarone by Alternaria longipes CGMCC 3.2875 was proposed.

关键词： Biotransformation α-Asarone Alternaria longipes CGMCC 3.2875 Anti-Aβ_(42)aggregation activity

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Resolvin D1 Induces mTOR-independent and ATG5-dependent Autophagy in BV-2 Microglial Cells

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Current Medical Science 2023年第6期43卷 1096-1106页

作者： Shang-wen PAN Li-sha HU Han WANG Rui-ting LI Ya-jun HE You SHANG Zhong-liang DAI Li-xin CHEN Wei XIONG Department of Critical Care medicine Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan430022China Department of Anesthesiology Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan430022China Department of Anesthesiology Qingdao Women and Children’s HospitalQingdao UniversityQingdao266034China Department of Anesthesiology The Second Clinical Medical College(Shenzhen People’s Hospital)Jinan UniversityShenzhen518020China Department of Pharmacology Medical CollegeJinan UniversityGuangzhou510632China integrated chinese and western medicine postdoctoral research station Jinan UniversityGuangzhou510632China

Objective The activation state of microglia is known to occupy a central position in the pathophysiological process of cerebral inflammation.Autophagy is a catabolic process responsible for maintaining cellular homeostasis.In recent years,autophagy has been demonstrated to play an important role in neuroinflammation.Resolvin D1(RvD1)is a promising therapeutic mediator that has been shown to exert substantial anti-inflammatory and proresolving activities.However,whether RvD1-mediated resolution of inflammation in microglia is related to autophagy regulation needs further investigation.The present study aimed to explore the effect of RvD1 on microglial autophagy and its corresponding pathways.Methods Mouse microglial cells(BV-2)were cultured,treated with RvD1,and examined by western blotting,confocal immunofluorescence microscopy,transmission electron microscopy,and flow cytometry.Results RvD1 promoted autophagy in both BV-2 cells and mouse primary microglia by favoring the maturation of autophagosomes and their fusion with lysosomes.Importantly,RvD1 had no significant effect on the activation of mammalian target of rapamycin(mTOR)signaling.Furthermore,RvD1-induced mTOR-independent autophagy was confirmed by observing reduced cytoplasmic calcium levels and suppressed calcium/calmodulin-dependent protein kinase II(CaMK II)activation.Moreover,by downregulating ATG5,the increased phagocytic activity induced by RvD1 was demonstrated to be tightly controlled by ATG5-dependent autophagy.Conclusion The present work identified a previously unreported mechanism responsible for the role of RvD1 in microglial autophagy,highlighting its therapeutic potential against neuroinflammation.

关键词： resolvin D1 microglia mTOR-independent autophagy ATG5-dependent autophagy phagocytosis

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Non-cytotoxic nanoparticles re-educating macrophages achieving both innate and adaptive immune responses for tumor therapy

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Asian Journal of Pharmaceutical Sciences 2022年第4期17卷 557-570页

作者： Shengmei Wang Xuanjun Liu Minghua Yang Linqi Ouyang Jinsong Ding Shengfeng Wang Wenhu Zhou Department of Pharmacy The Third Xiangya HospitalCentral South UniversityChangshaHunan410013China Xiangya School of Pharmaceutical Sciences Central South UniversityChangshaHunan410013China postdoctoral research station of Clinical medicine and Department of Pediatrics The Third Xiangya HospitalCentral South UniversityChangshaHunan410013China The First Hospital of Hunan University of chinese medicine ChangshaHunan410007China

Macrophages are important antigen-presenting cells to combat tumor via both innate and adaptive immunity,while they are programmed toM2 phenotype in established tumors and instead promote cancer development and metastasis.Here,we develop a nanomedicine that can re-educate M2 polarized macrophages to restore their anti-tumor activities.The nanomedicine has a core-shell structure to co-load IPI549,a PI3Kγinhibitor,and CpG,a Toll-like receptor 9 agonist.Specifically,the hydrophobic IPI549 is self-assembled into a pure drug nano-core,while MOF shell layer is coated for CpG encapsulation,achieving extra-high total drugs loading of 44%.Such nanosystem could facilitate intracellular delivery of the payloads but without any cytotoxicity,displaying excellent biocompatibility.After entering macrophages,the released IPI549 and CpG exert a synergistic effect to switch macrophages from M2 to M1 phenotype,which enables anti-tumor activities via directly engulfing tumor cells or excreting tumor killing cytokines.Moreover,tumor antigens released from the dying tumor cells could be effectively presented by the re-educated macrophages owing to the up-regulation of various antigen presenting mediators,resulting in infiltration and activation of cytotoxic T lymphocytes.As a result,the nanosystem triggers a robust antitumor immune response in combination with PD-L1 antibody to inhibit tumor growth and metastasis.This work provides a non-cytotoxic nanomedicine to modulate tumor immune microenvironment by reprograming macrophages.

关键词： Core-shell nanoparticles Metal-organic-framework CpG IPI549 Tumor immunotherapy

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Network pharmacology research of Chuanxiong Rhizoma-Acori Tatarinowii Rhizoma in the treatment of vascular dementia

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Aging Communications 2022年第1期4卷 28-36页

作者： Dan Liu Si-Miao Ran Shuo Yang Zhi-Zhong Ma He-Yuan Shi Ai-Hua Tan Zheng Zhang HuangGang Hospital of TCM Affiliated to Hubei University of chinese medicine HuangGang 438000China Institute of Geriatrics Hubei University of Chinese MedicineWuhan 430060China Department of integrated chinese and western medicine School of Basic Medical SciencesPeking UniversityBeijing 100083China postdoctoral research station Beijing University of Chinese MedicineBeijing 100190China

Objective:Using network pharmacology to explore the target and mechanism of Chuanxiong Rhizoma and Acori Tatarinowii Rhizoma in the treatment of vascular dementia(VaD),so as to provide a reference for treating VaD through them.Methods:Traditional chinese medicine systems pharmacology database and analysis platform were used to screen the main active ingredients and targets of Chuanxiong Rhizoma and Acori Tatarinowii Rhizoma.By means of Gene Cards and Online Mendelian Inheritance in Man(OMIM),targets of VaD were collected.The intersecting targets were obtained by using the Venn map.The String online database was used to build a protein-protein interactions Network and the Metascape database was used to perform GO function enrichment analysis and KEGG pathway enrichment analysis.A“drug-ingredient-target-pathway”network was constructed by Cytoscape software.Autodock vina software was used to conduct molecular docking between targets.Results:A total of 7 active ingredients in Chuanxiong Rhizoma and 4 active ingredients in Acori Tatarinowii Rhizoma were screened.There were 42 active targets of Chuanxiong Rhizoma and 70 active targets of Acori Tatarinowii Rhizoma and 1152 disease targets.After deleting the repeat value,51 drugs targets were obtained.After the intersection,with a total of 25 targets.According to GO and KEGG enrichment analysis,the main biological processes involved include cellular response to lipid,negative regulation of apoptotic signaling pathway,blood circulation,response to a steroid hormone,etc.The main pathways include pathways in cancer,PI3K-Akt signaling pathway,and AGE-RAGE signaling pathway in diabetic complications,etc.Molecular docking showed that the most active docking combinations were AKT1 and Perlolyrine,RELA and FA,MAPK14 and FA,respectively.Conclusion:Chuanxiong Rhizoma and Acori Tatarinowii Rhizoma play an important role in the treatment of VaD mainly by anti-inflammatory and anti-apoptosis.

关键词： Chuanxiong Rhizoma Acori Tatarinowii Rhizoma Vascular Dementia network pharmacology

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Role of Heteroplasmic Mutations in the Mitochondrial Genome and the ID4 Gene Promoter Methylation Region in the Pathogenesis of Chronic Aplastic Anemia in Patients Suffering from Kidney Yin Deficiency

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chinese Journal of Integrative medicine 2016年第6期22卷 412-419页

作者：崔兴王敬毅刘奎崔思远张杰罗雅琴王欣 Department of Hematology Affiliated Hospital of Shandong University of Traditional Chinese Medicine postdoctoral research station Shandong University Department of Hematology Shangdong Province Hospital of Shandong University

Objective： To analyze changes in gene amplification in the mitochondrial genome and in the ID4 gene promoter methylation region in patients with chronic aplastic anemia （CAA） suffering from Kidney （Shen） yin deficiency or Kidney yang deficiency. Methods： Bone marrow and oral epithelium samples were collected from CAA patients with Kidney yin deficiency or Kidney yang deficiency （20 cases）. Bone marrow samples were collected from 20 healthy volunteers. The mitochondrial genome was amplified by polymerase chain reaction （PCR）, and PCR products were used for sequencing and analysis. Results： Higher mutational rates were observed in the ND1-2, ND4-6, and CYTB genes in CAA patients suffering from Kidney yin deficiency. Moreover, the ID4 gene was unmethylated in bone marrow samples from healthy individuals, but was methylated in some CAA patients suffering from Kidney yin deficiency （positive rate, 60%） and Kidney yang deficiency （positive rate, 55%）. Conclusions： These data supported that gene mutations can alter the expression of respiratory chain enzyme complexes in CAA patients, resulting in energy metabolism impairment and promoting the physiological and pathological processes of hematopoietic failure. Functional impairment of the mitochondrial respiration chain induced by gene mutation may be an important reason for hematopoietic failure in patients with CA,＆,. This change is closely related to maternal inheritance and Kidney yin deficiency. Finally, these data supported the assertion that it is easy to treat disease in patients suffering from yang deficiency and difficult to treat disease in patients suffering from yin deficiency.

关键词： chronic aplastic anemia Kidney bone marrow mitochondrial DNA maternal inheritance ID4 methylation

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Inhibitive effect on phagocytosis of Candida albicans induced by pretreatment with quercetin via actin cytoskeleton interference

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Journal of Traditional chinese medicine 2013年第6期33卷 804-809页

作者： Shuna Cui Jing Qian Ping Bo Department of integrated Traditional chinese Medicne and western medicine Medical College of Yangzhou University Biologic Systems Analysis Group Helmholtz Centre for Infection Research Department of integrated TCM and western medicine Medical College of Yangzhou University

OBJECTIVE: To investigate the anti-inflammatory effect of pretreatment with quercetin on macrophages after Candida albicans infection. METHODS: RAW 264.7 macrophages were used as a target cell line. Cell viability after treatment with quercetin at different time points was detected by Carboxyfluorescein diacetate, succinimidyl ester. Phagocytic function of macrophages was determined by a fluorometric assay. Cytokine tumor necrosis factor α(TNF-α) production was measured by enzyme-linked immunosorbent assay. F-actin cytoskeleton of L929 cells was stained by Alexa Fluor 488-phalloidin. RESULTS: Pretreatment with quercetin decreased cell viability only at the highest concentration of 37 μg/mL 2, 24, and 48 h after the treatment. The phagocytic efficiency of macrophages pretreated with quercetin was significantly decreased in a timeand dose-dependent manner. F-actin labeling showed that the actin cytoskeleton of the cells started to break down 2 h after treatment. Moreover, it notably inhibited cytokine TNF-α production after Candida albicans infection. CONCLUSION: Pretreatment with quercetin induced an anti-inflammatory effect against Candida albicans infection in macrophages.

关键词：肌动蛋白细胞骨架白色念珠菌吞噬功能槲皮素预处理抑制作用诱导酶联免疫吸附法

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New ent-kaurane diterpenes with chiral epoxyangelate moieties from Wedelia prostrata

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chinese Chemical Letters 2019年第2期30卷 451-453页

作者： Zhongnan Wu Yubo Zhang Wen Li Nenghua Chen Qianwen Niu Yingying Li Qingguo Li Dan Yang Yaolan Li Guocai Wang Institute of Traditional chinese medicine ξ Natural Products College of Pharmacy and Guangdong Province Key Laboratory of Pharmacodynamic Constituents of TCM and New Drugs ResearchJinan University integrated chinese and western medicine postdoctoral research station Jinan University School of Pharmaceutical Sciences Guangzhou University of Chinese Medicine Department of Specific Diagnosis The 324th Military Hospital

Four new ent-kaurane diterpenes with chiral epoxyangelate moieties, (2′R,3′R)-3 a- (2′,3′-epoxyangeloyloxy)-kaur-16-en-19-oic acid (1), (2′S,3′S)-3 a- (2′,3′-epoxyangeloyloxy)-kaur-16-en-19-oic acid (2), (2′S,3′R)-3 a- (2',3'-epoxyangeloyloxy)-kaur-16-en-19-oic acid (3) and (2′R,3′S)-3α- (2′,3'-epoxyangeloyloxy)-kaur-16-en-19-oic acid (4), along with eight known diterpenes (5-12), were isolated from Wedelia prostrata. The absolute configurations of the new structures were determined by X-ray crystallography,ECD calculations and chemical methods. All compounds were evaluated for their cytotoxicity activities on human HepG-2 cells,with IC_(50) values of 11.72 ±0.22 μmol/L to 54.75±1.12 μmol/L.

关键词： Wedelia prostrata Chiral epoxyangelate moieties Ent-kaurane diterpenes Absolute configuration Cytotoxicity activities

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