Thiostrepton(TST)is a natural antibiotic with pleiotropic properties.This study aimed to elucidate the therapeutic effect of TST on experimental colitis and identify its targets.The effect of TST on colon inflammation...Thiostrepton(TST)is a natural antibiotic with pleiotropic properties.This study aimed to elucidate the therapeutic effect of TST on experimental colitis and identify its targets.The effect of TST on colon inflammation was evaluated in a dextran sulfate sodium(DSS)-induced colitis model and a T-cell transfer colitis model.The therapeutic targets of TST were investigated by cytokine profiling,immunophenotyping and biochemical approaches.The effect of TST on the gut microbiota and its contribution to colitis were evaluated in mice with DSS-induced colitis that were subjected to gut microbiota depletion and fecal microbiota transplantation(FMT).Alterations in the gut microbiota caused by TST were determined by 16S rDNA and metagenomic sequencing.Here,we showed that TST treatment significantly ameliorated colitis in the DSS-induced and T-cell transfer models.Specifically,TST targeted the retinoic acid-related orphan nuclear receptor RORγt to reduce the production of IL-17A byγδT cells,type 3 innate lymphoid cells(ILC3s)and Th17 cells in mice with DSS-induced colitis.Similarly,TST selectively prevented the development of Th17 cells in the T-cell transfer colitis model and the differentiation of naïve CD4^(+)T cells into Th17 cells in vitro.Mechanistically,TST induced the ubiquitination and degradation of RORγt by promoting the binding of Itch to RORγt.Moreover,TST also reversed dysbiosis to control colonic inflammation.Taken together,these results from our study describe the previously unexplored role of TST in alleviating colonic inflammation by reducing IL-17A production and modulating dysbiosis,suggesting that TST is a promising candidate drug for the treatment of IBD.展开更多
A high-expression system of L11 was constructed and investigated its interaction with other elements of the ribosome using physicochemical methods. The gene rplK, coding for the protein L11 from the E. coli 50S riboso...A high-expression system of L11 was constructed and investigated its interaction with other elements of the ribosome using physicochemical methods. The gene rplK, coding for the protein L11 from the E. coli 50S ribosomal subunit was amplifyied, cloned and over-expressed. The protein L11 was purified under native and denaturing conditions, refolded and the structure of both proteins was compared. The protein L11 properly refolded from 6M urea after dialysis. Experiments on binding of proteins L11, RRF and EF-G from Escherichia coli were performed by ana-lytical centrifugation and Biacore. Specific binding between protein L11 and RRF by analytical cen-trifugation was not detected probably due to struc-tural reasons. These findings may be helpful in the design of new antibiotics that specifically disrupt the interactions in the “GTP-associated site” of the bac-terial ribosome, as many of them are not effective anymore. A common intrinsically disordered region of protein L11 was found to be the amino acid se-quence 86-97, while the residues 67-74, containing the linker region, are predicted to be disordered by DisEMBL.展开更多
目的研究硫链丝菌肽(TST)对人鼻咽癌HNE-1细胞株增殖能力及Forkhead box protein M1(Foxm1)表达的影响。方法以MTT法分别测定不同浓度(1、2、4、8、12、16μmol/L)TST作用于人鼻咽癌HNE-1细胞48h的增殖抑制率;通过流式细胞仪(FCM)检测...目的研究硫链丝菌肽(TST)对人鼻咽癌HNE-1细胞株增殖能力及Forkhead box protein M1(Foxm1)表达的影响。方法以MTT法分别测定不同浓度(1、2、4、8、12、16μmol/L)TST作用于人鼻咽癌HNE-1细胞48h的增殖抑制率;通过流式细胞仪(FCM)检测不同浓度(1、2μmol/L)TST分别作用于HNE-1细胞48h后对其细胞周期动力学的影响;采用免疫细胞化学法和Western blot检测HNE-1细胞中Foxm1蛋白表达以及不同浓度TST作用于HNE-1细胞48hFoxm1蛋白表达的变化。结果不同浓度(1、2、4、8、12、16μmol/L)TST作用于HNE-1细胞48h后的增殖抑制率分别为8.31%、17.46%、34.28%、58.68%、87.91%、99.05%;浓度分别为1、2μmol/L的TST作用于HNE-1细胞48h后,均使HNE-1细胞停滞于G1/G0期(P<0.05),且呈剂量依赖性(P<0.05);同时将TST作用于HNE-1细胞48后,经免疫细胞化学法和Western blot均检测到Foxm1蛋白在HNE-1细胞胞质中表达显著降低(P<0.05)。结论 TST对人鼻咽癌HNE-1细胞株具有明显的增殖抑制作用,可能与降低肿瘤细胞中Foxm1蛋白表达有关。展开更多
基金This work was supported by the National Key Research and Development Program of China(2018YFA0507900)the National Natural Science Foundation of China(81802460)the Natural Science Foundation of Chongqing(CSTB2022NSCQ-MSX0184).
文摘Thiostrepton(TST)is a natural antibiotic with pleiotropic properties.This study aimed to elucidate the therapeutic effect of TST on experimental colitis and identify its targets.The effect of TST on colon inflammation was evaluated in a dextran sulfate sodium(DSS)-induced colitis model and a T-cell transfer colitis model.The therapeutic targets of TST were investigated by cytokine profiling,immunophenotyping and biochemical approaches.The effect of TST on the gut microbiota and its contribution to colitis were evaluated in mice with DSS-induced colitis that were subjected to gut microbiota depletion and fecal microbiota transplantation(FMT).Alterations in the gut microbiota caused by TST were determined by 16S rDNA and metagenomic sequencing.Here,we showed that TST treatment significantly ameliorated colitis in the DSS-induced and T-cell transfer models.Specifically,TST targeted the retinoic acid-related orphan nuclear receptor RORγt to reduce the production of IL-17A byγδT cells,type 3 innate lymphoid cells(ILC3s)and Th17 cells in mice with DSS-induced colitis.Similarly,TST selectively prevented the development of Th17 cells in the T-cell transfer colitis model and the differentiation of naïve CD4^(+)T cells into Th17 cells in vitro.Mechanistically,TST induced the ubiquitination and degradation of RORγt by promoting the binding of Itch to RORγt.Moreover,TST also reversed dysbiosis to control colonic inflammation.Taken together,these results from our study describe the previously unexplored role of TST in alleviating colonic inflammation by reducing IL-17A production and modulating dysbiosis,suggesting that TST is a promising candidate drug for the treatment of IBD.
文摘A high-expression system of L11 was constructed and investigated its interaction with other elements of the ribosome using physicochemical methods. The gene rplK, coding for the protein L11 from the E. coli 50S ribosomal subunit was amplifyied, cloned and over-expressed. The protein L11 was purified under native and denaturing conditions, refolded and the structure of both proteins was compared. The protein L11 properly refolded from 6M urea after dialysis. Experiments on binding of proteins L11, RRF and EF-G from Escherichia coli were performed by ana-lytical centrifugation and Biacore. Specific binding between protein L11 and RRF by analytical cen-trifugation was not detected probably due to struc-tural reasons. These findings may be helpful in the design of new antibiotics that specifically disrupt the interactions in the “GTP-associated site” of the bac-terial ribosome, as many of them are not effective anymore. A common intrinsically disordered region of protein L11 was found to be the amino acid se-quence 86-97, while the residues 67-74, containing the linker region, are predicted to be disordered by DisEMBL.
基金National Natural Science Foundation of China(No.31100042)Guangdong Natural Science Foundation(No.S2011010001625)+1 种基金Guangzhou Pearl River Rising Star Program for Science and Technology(No.2013091)Guangdong Chinese Academy of Science Comprehensive Strategic Cooperation Project(No.2012B091100276)
