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Tanshinone IIA protects intestinal epithelial cells from ferroptosis through the upregulation of GPX4 and SLC7A11

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BIOCELL 2023年第5期47卷 1107-1115页

作者： HAN WANG YANG SUN XIAOXU ZHANG XIAOYING WANG YUJUN XIA LISHENG WANG School of Basic Medicine Qingdao UniversityQingdao266071China Laboratory of Molecular Diagnosis and Regenerative Medicine The Affiliated Hospital of Qingdao UniversityQingdao266000China Department of Hematology The Affiliated Hospital of Qingdao UniversityQingdao266000China Beijing Institute of Radiation Medicine Beijing100850China

Background:Inflammatory bowel disease(IBD)is a chronic inflammatory disease of the gastrointestinal tract.The destruction of the intestinal epithelial barrier is one of the major pathological processes in IBD pathology.Growing evidence indicated that epithelial cell ferroptosis is linked to IBD and is considered a target process.Methods:RAS-selective lethal 3(RSL3)was used to induce ferroptosis in intestinal epithelial cell line No.6(IEC-6)cells,and cell ferroptosis and the effects of tanshinone IIA(Tan IIA)were determined by cell counting kit-8(CCK-8),reactive oxygen species(ROS)staining,Giemsa staining and transmission electron microscope(TEM).The cell viability of natural product library compounds was determined by CCK-8.The expression of ferroptosis-related genes were detected by real-time quantitative polymerase chain reaction(RT-qPCR)and western blot.Results:Treatment of IEC-6 cells results in the accumulation of ROS and typical morphological characteristics of ferroptosis.RSL3 treatment caused rapid cellular cytotoxicity which could be reversed by ferrostatin-1(Fer-1)in IEC-6 cells.Natural product library screening revealed that Tan IIA is a potent inhibitor of IEC-6 cell ferroptosis.Tan IIA could significantly protect the RSL3-induced ferroptosis of IEC-6 cells.Furthermore,the ferroptosis suppressors,glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(SLC7A11),and miR-17-92 were found to be early response genes in RSL3-treated cells.Treatment of IEC-6 cells with Tan IIA resulted in upregulation of GPX4,SLC7A11,and miR-17-92.Conclusion:Our study demonstrated that Tan IIA protects IEC-6 cells from ferroptosis through the upregulation of GPX4,SLC7A11,and miR-17-92.The findings might provide a theoretical grounding for the future application of Tan IIA to treat or prevent IBD.

关键词： Tanshinone IIA GPX4 Ferroptosis intestinal epithelial cells IBD

A Comparative Study on Rat intestinal epithelial cells and Resident Gut Bacteria (ii) Effect of Arsenite

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Biomedical and Environmental Sciences 2006年第2期19卷 77-86页

作者： RAJ K.UPRETI A.KANNAN RICHA SHRIVASTAVA U.C.CHATURVEDI Biomembrane Toxicology Laboratory Industrial Toxicology Research Centre M. G. Marg P. B. No. 80 Lucknow-226001 India

Objective In order to use facultative gut bacteria as an alternate to animals for the initial gastrointestinal toxicity screening of heavy metals, a comparative study on rat intestinal epithelial cells and resident gut bacteria was undertaken. Methods in vitro growth rate of four gut bacteria, dehydrogenase （DHA） and esterase （EA） activity test, intestinal epithelial and bacterial cell membrane enzymes and in situ effect of arsenite were analysed. Results Growth profile of mixed resident population of gut bacteria and pure isolates of Escherichia coli, Pseudomonas sp., Lactobacillus sp., and Staphylococcus sp. revealed an arsenite （2-20 ppm） concentration-dependent inhibition. The viability pattern of epithelial cells also showed similar changes. DHA and EA tests revealed significant inhibition （40%-72%） with arsenite exposure of 5 and 10 ppm in isolated gut bacteria and epithelial cells. Decrease in membrane alkaline phosphatase and Ca^2＋-Mg^2＋-ATPase activities was in the range of 33%-55% in four bacteria at the arsenite exposure of 10 ppm, whereas it was 60%-65% in intestinal epithelial villus cells, in situ incubation of arsenite using intestinal loops also showed more or less similar changes in membrane enzymes of resident gut bacterial population and epithelial cells. Conclusion The results indicate that facultative gut bacteria can be used as suitable in vitro model for the preliminary screening of arsenical gastrointestinal cytotoxic effects.

关键词： Arsenic Gut microflora intestinal bacteria intestinal epithelial cells Membrane enzymes GI-toxicity

Proteomic analysis of the changed proteins of Trichinella spiralis infective larvae after co-culture in vitro with intestinal epithelial cells

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Proteomic analysis of the changed proteins of Trichinella sp...

Microbial sensing in the intestine

中国动物学会寄生虫学专业委员会第十三次全国学术会议暨第...

作者： Lei Wang Zhong-Quan Wang Jing Cui Department of Parasitology Medical College Zhengzhou University

The purpose of this study was to determine the protein changes of Trichinella spiralis infective larvae after co-culture in vitro with intestinal epithelial cells and to screen the proteins related with invasion.The infective larvae were incubated with HCT-8 cells for 18h;the larval soluble proteins were analyzed by SDS-PAGE and Western blot.Results showed after culture with HCT-8 cells,seven additional protein bands（123,77,58,36,30,28 and 21 k Da）of the larvae were recognized by sera of the infected mice,which may be the invasion-related proteins released by the infective larvae;Three protein bands（97,51,23 k Da）were not recognized by the above sera,which may be enzymolyzed by enzymes released from HCT-8 cells,compared with proteins of the larvae incubated only in RPMI-1640 medium.Three bands（58,36,21k Da）of seven discrepant protein bands were excised from gels,in-gel digested with trypsin,and identified by liquid chromatography and tandem mass spectrometry（LC-MS/MS）combined with bioinformatics.A total of 211proteins were identified.Numerous enzymes（e.***.metallo-and serine proteases,cysteine proteinases,5’-nucleotidase,enolase）were found to be highly expressed,which may play significant role in invasion.The identification of these larval proteins represents the first step to elucidate their specific roles on host-parasite relationship.Results suggested that this proteomic approach was a useful tool to study protein expression in T.spiralis and will contribute to the identification of the proteins related with invasion.

关键词： Trichinella spiralis invasion intestinal epithelial cells proteomics mass spectrometry

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Protein & Cell 2023年第11期14卷 824-860页

作者： Tingting Wan Yalong Wang Kaixin He Shu Zhu Division of Life Sciences and Medicine The CAS Key Laboratory of Innate Immunity and Chronic DiseaseInstitute of ImmunologySchool of Basic Medical SciencesUniversity of Science and Technology of ChinaHefei 230027China Department of Digestive Disease Division of Life Sciences and MedicineThe First Affiliated Hospital of USTCUniversity of Science and Technology of ChinaHefei 230001China Institute of Health and Medicine Hefei Comprehensive National Science CenterHefei 230601China

The gut microbiota plays a key role in host health and disease,particularly through their interactions with the immune system.intestinal homeostasis is dependent on the symbiotic relationships between the host and the diverse gut microbiota,which is influenced by the highly co-evolved immune-microbiota interactions.The first step of the interaction between the host and the gut microbiota is the sensing of the gut microbes by the host immune system.In this review,we describe the cells of the host immune system and the proteins that sense the components and metabolites of the gut microbes.We further highlight the essential roles of pattern recognition receptors(PRRs),the G protein-coupled receptors(GPCRs),aryl hydrocarbon receptor(AHR)and the nuclear receptors expressed in the intestinal epithelial cells(IECs)and the intestine-resident immune cells.We also discuss the mechanisms by which the disruption of microbial sensing because of genetic or environmental factors causes human diseases such as the inflammatory bowel disease(IBD).

关键词： mucosal immunology pattern recognition receptors protein-coupled receptors intestinal epithelial cells immune cells gut microbiota enteric viruses inflammatory bowel disease

Mannose metabolism normalizes gut homeostasis by blocking the TNF-α-mediated proinflammatory circuit

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Cellular & Molecular Immunology 2023年第2期20卷 119-130页

作者： Peng Xiao Ziwei Hu Jiaheng Lang Tianyuan Pan Randall Tyler Mertens Huilun Zhang Ke Guo Manlu Shen Hongqiang Cheng Xue Zhang Qian Cao Yuehai Ke Department of Gastroenterology Sir Run Run Shaw HospitalZhejiang University School of MedicineHangzhouChina Department of Pathology and Pathophysiology and Department of Gastroenterology at Sir Run Run Shaw HospitalZhejiang University School of MedicineHangzhouChina Immunological Disease Research Center Sir Run Run Shaw HospitalZhejiang University School of MedicineHangzhouChina Key Laboratory for Immunity and Inflammatory Diseases of Zhejiang Province HangzhouChina Department of Pathology and Pathophysiology Zhejiang University School of MedicineHangzhouChina Department of General Medicine The First Affiliated HospitalZhejiang University School of MedicineHangzhouChina Department of Immunology Harvard Medical SchoolBostonMAUSA Zhejiang Laboratory for Systems and Precision Medicine Zhejiang University Medical CenterHangzhouChina

Mannose is a naturally occurring sugar widely consumed in the daily diet;however,mechanistic insights into how mannose metabolism affects intestinal inflammation remain lacking.Herein,we reported that mannose supplementation ameliorated colitis development and promoted colitis recovery.Macrophage-secreted inflammatory cytokines,particularly TNF-α,induced pathological endoplasmic reticulum stress(ERS)in intestinal epithelial cells(IECs),which was prevented by mannose via normalization of protein N-glycosylation.By preserving epithelial integrity,mannose reduced the inflammatory activation of colonic macrophages.On the other hand,mannose directly suppressed macrophage TNF-αproduction translationally by reducing the glyceraldehyde 3-phosphate level,thus promoting GAPDH binding to TNF-αmRNA.Additionally,we found dysregulated mannose metabolism in the colonic mucosa of patients with inflammatory bowel disease.Finally,we revealed that activating PMM2 activity with epalrestat,a clinically approved drug for the treatment of diabetic neuropathy,elicited further sensitization to the therapeutic effect of mannose.Therefore,mannose metabolism prevents TNF-α-mediated pathogenic crosstalk between IECs and intestinal macrophages,thereby normalizing aberrant immunometabolism in the gut.

关键词： Colitis Mannose Macrophages intestinal epithelial cells TNF-alpha Endoplasmic reticulum stress

Crosstalk Between epithelial cells and Dendritic cells in intestinal Homeostasis

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Crosstalk Between Epithelial Cells and Dendritic Cells in In...

Phytoglycoprotein isolated from Dioscorea batatas Decne promotes intestinal epithelial wound healing

第九届全国免疫学学术大会

作者： Mei Liu Peng Wang Dongyan Liu Shengjing Hospital of China Medical University

The human intestine covers a huge surface area of approximately 200-400 m.Residing within the lumen are an estimated 100 trillion microbial cells.The gastrointestinal tract represents the largest mucosal membrane surface,which is one of the most complex organs of the human body.The mucosal immune system has the very difficult task to protect against invaders and to promote tolerance toward food antigens and the microbiota.These activities are achieved via a complex interaction between immune cells and the local microenvironment.intestinal epithelial cells(IECs)play an important role in intestinal inflammation infection and cancer development.IECs not only constitute the first barrier in the gut,but also are crucial for integrating the external and internal signals and for coordinating the ensuing immune response.Dendritic cells(DCs)are specialized antigen-presenting cells,because of their high plasticity and maturation ability in response to local danger signals derived from innate immunity,DCs are key actors in the connection between innate immunity and adaptive immunity responses.The intestinal epithelial cells coordinate with the DCs in immune recognition,tolerance and host defense.In this review,We will summarize the crosstalk between the IECs and intestinal DCs in gut homeostasis and diseases.

关键词： intestinal epithelial cells Dendritic cells Microbiota

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Chinese Journal of Natural Medicines 2020年第10期18卷 738-748页

作者： KIM Ji-Yun LEE Young-Min PARK Jong-Pil LIM Kye-Taek LEE Sei-Jung Department of Pharmaceutical Engineering Daegu Haany UniversityGyeongsan 38610Republic of Korea Department of Food Science and Technology Chung-Ang UniversityAnseong 17546Republic of Korea Division of Animal Science Chonnam National UniversityGwangju61186Republic of Korea

Dioscorea batatas Decne(DBD)has been used to heal various illnesses of the kidney and intestine as an herbal medicine in Asia.As a source of therapeutic agents,many glycoproteins have been isolated from mushrooms and plants,but the functional role of glycoprotein in intestinal epithelial wound healing has not been understood yet.In the present study,we investigated the wound healing potentials of the 30 kDa glycoprotein(DBD glycoprotein)isolated from DBD in human intestinal epithelial(INT-407)cells.We found that DBD glycoprotein(100μg·mL^-1)significantly increased the motility of INT-407 cells for 24 h by activating protein kinase C(PKC).DBD glycoprotein stimulated the activation of p38 mitogen-activated protein kinase(MAPK),which is responsible for the phosphorylation of NF-κB inhibitorα(IκBα).DBD glycoprotein increased the level of profilin-1(PFN1),α-actinin and F-actin expression via activation of transcription factor,nuclear factor-kappa B(NF-κB)during its promotion of cell migration.Experimental mouse colitis was induced by adding dextran sulfate sodium(DSS)to the drinking water at a concentration of 4%(W/V)for 7 days.We figured out that administration of DBD glycoprotein(10 and 20 mg·kg-1)lowers the levels of disease activity index and histological inflammation in DSS-treated ICR mice.In this regard,we suggest that DBD glycoprotein has ability to promote the F-actin-related migration signaling events via activation of PKC and NF-κB in intestinal epithelial cells and prevent inflammatory bowel disease.

关键词： Cell migration Dioscorea batatas Decne(DBD)glycoprotein intestinal epithelial cells p38 MAPK F-actin Mouse colitis

Effect of EPEC endotoxin and bifidobacteria on intestinal barrier function through modulation of toll-like receptor 2 and toll-like receptor 4 expression in intestinal epithelial cell-18

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World Journal of Gastroenterology 2017年第26期23卷 4744-4751页

作者： Xia Yang Xian-Chun Gao Jun Liu Hong-Yu Ren Division of Gastroenterology Union Hospital Tongji Medical College Huazhong University of Science and Technology

AIM To investigate toll-like receptor 2(TLR2) and TLR4 expression, following bifidobacteria and low-dose EPEC endotoxin treatment, and intestinal barrier function in rat intestinal epithelial cell-18(IEC-18).METHODS Six experimental groups were established-normal control, EPEC, Bifidobacteria infantis(B. infantis), B. longum, B. bifidum, and B. youth groups. Optimal EPEC endotoxin concentration, bifidobacteria fold dilution, and treatment duration were determined. Quantitative real-time polymerase chain reaction and western blot, respectively, were conducted to detect TLR2 and TLR4 m RNA and protein expression in IEC-18 cells. Transepithelial electrical resistance(TEER) was measured by the EVOM chopstick voltohmmeter in each group. All experiments were conducted in triplicate and data were analyzed on SPSS 16.RESULTS TLR2 and TLR4 m RNA and protein expression in the EPEC group were significantly higher than in the control group(P cells against injury by down-regulating TLR2 and TLR4 expression and enhance intestinal barrier function to protect the intestinal epithelial cells from pathogenic invasion.

关键词： Bifidobacteria intestinal barrier function intestinal epithelial cells

Chang'an Ⅱ Decoction(肠安Ⅱ号方)-Containing Serum Ameliorates Tumor Necrosis Factor-α-Induced intestinal epithelial Barrier Dysfunction via MLCK-MLC Signaling Pathway in Rats

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Chinese Journal of Integrative Medicine 2020年第10期26卷 745-753页

作者： CHEN Ting YIN Xiao-Ian KANG Nan WANG Xiao-ge LI Bao-shuang JI Hai-jie ZHANG Yin-qiang BIAN Li-qun ZHANG Bei-hua WANG Feng-yun TANG Xu-dong Department of Gastroenterology Xiyuan HospitalChina Academy of Chinese Medical SciencesBeijing(100091)China Graduate School of China Academy of Chinese Medical Sciences Beijing(100700)China Department of Internal Medicine Jining Medical College Affiliated HospitalJining(272029)Shandong ProvineeChina Department of Gastroenterology Henan University of Chinese Medicine Affiliated HospitalZhengzhou(136300)China Department of Liver Diseases Xiyuan HospitalChina Academy of Chinese

Objective To investigate the effect of Chang’an Ⅱ Decoction(肠安Ⅱ号方))-containing serum on intestinal epithelial barrier dysfunction in rats.Methods Tumor necrosis factor(TNF)-α-induced injury of Caco-2 monolayers were established as an inflammatory model of human intestinal epithelium.Caco-2 monolayers were treated with blank serum and Chang’an Ⅱ Decoction-containing serum that obtained from the rats which were treated with distilled water and Chang’an Ⅱ Decoction intragastrically at doses of 0.49,0.98,1.96 g/(kg·d)for 1 week,respectively.After preparation of containing serum,cells were divided into the normal group,the model group,the Chang’an Ⅱ-H,M,and L groups(treated with 30 ng/mL TNF-αand medium plus 10%high,middle-,and low-doses Chang’an Ⅱ serum,respectively).epithelial barrier function was assessed by transepithelial electrical resistance(TER)and permeability of fluorescein isothiocyanate(FITC)-labeled dextran.Transmission electron microscopy was used to observe the ultrastructure of tight junctions(TJs).Immunofluorescence of zonula occludens-1(ZO-1),claudin-1 and nuclear transcription factor-kappa p65(NF-κBp65)were measured to determine the protein distribution.The mRNA expression of myosin light chain kinase(MLCK)was measured by real-time polymerase chain reaction.The expression levels of MLCK,myosin light chain(MLC)and p-MLC were determined by Western blot.Results Chang’an Ⅱ Decoction-containing serum significantly attenuated the TER and paracellular permeability induced by TNF-α.It alleviated TNF-α-induced morphological alterations in TJ proteins.The increases in MLCK mRNA and MLCK,MLC and p-MLC protein expressions induced by TNF-αwere significantly inhibited in the Chang’an Ⅱ-H group.Additionally,Chang’an Ⅱ Decoction significantly attenuated translocation of NF-κBp65 into the nucleus.Conclusion High-dose Chang’an Ⅱ-containing serum attenuates TNF-α-induced intestinal barrier dysfunction.The underlying mechanism may be involved in inhibiting the MLCK-MLC phosphorylation signaling pathway mediated by NF-κ

关键词： myosin light chain kinase-myosin light chain signaling pathway intestinal epithelial cells tight junction tumor necrosis factor-α Chang’an II Decoction drug-containing serum