妊娠期糖尿病(GDM)是妊娠期间常见的糖代谢异常疾病。本文探讨了吲哚胺2,3-双加氧酶(IDO)与人类白细胞抗原-G (HLA-G)两种免疫调节分子在GDM中的表达及临床意义。本文指明IDO在妊娠期间的活性增加有助于保护胎儿免受母体免疫系统的攻击...妊娠期糖尿病(GDM)是妊娠期间常见的糖代谢异常疾病。本文探讨了吲哚胺2,3-双加氧酶(IDO)与人类白细胞抗原-G (HLA-G)两种免疫调节分子在GDM中的表达及临床意义。本文指明IDO在妊娠期间的活性增加有助于保护胎儿免受母体免疫系统的攻击,但在GDM条件下会造成IDO活性降低。HLA-G通过抑制免疫细胞功能维持母胎间免疫平衡,但GDM会导致血清中可溶性HLA-G水平下降甚至变异。IDO活性降低和HLA-G表达变化提示免疫耐受受损,联合检测这两种分子可为GDM的早期诊断、病情监控及治疗评估提供重要信息。Gestational Diabetes Mellitus (GDM) is a common disorder of glucose metabolism during pregnancy. This article explores the expression and clinical significance of two immunomodulatory molecules, Indoleamine 2,3-Dioxygenase (IDO) and Human Leukocyte Antigen-G (HLA-G), in GDM. It is indicated that the increased activity of IDO during pregnancy helps protect the fetus from maternal immune system attacks, but under GDM conditions, it leads to a reduction in IDO activity. HLA-G maintains immune balance between mother and fetus by inhibiting immune cell function, but GDM causes a decrease or even variation in the levels of soluble HLA-G in the serum. The reduction in IDO activity and changes in HLA-G expression suggest impaired immune tolerance. The combined detection of these two molecules can provide crucial information for the early diagnosis, monitoring, and treatment evaluation of GDM.展开更多
Objectives:Triple-negative breast cancer(TNBC)is the breast cancer subtype with the poorest prognosis.This study aimed to elucidate the molecular pathways through which isoliquiritigenin(ISL),a natural chalcone compou...Objectives:Triple-negative breast cancer(TNBC)is the breast cancer subtype with the poorest prognosis.This study aimed to elucidate the molecular pathways through which isoliquiritigenin(ISL),a natural chalcone compound derived fromlicorice and other plant roots,targets interferon regulatory factor 5(IRF5)in TNBC.Methods:TNBC cell lines were cultured and subjected to IRF5 knockdown using short hairpin RNA.Cell proliferation was assessed by cell counting kit-8(CCK-8)assay and colony formation assays.Western blotting and quantitative reverse transcription polymerase chain reaction(RT-PCR)were employed to measure expression levels of IRF5,solute carrier family 7 member 5(SLC7A5),and indoleamine 2,3-dioxygenase 1(IDO1).Intracellular tryptophan and its metabolites were quantified using commercially available assay kits and high-performance liquid chromatography(HPLC).TNBC cells were treated with various concentrations of ISL to evaluate its effects on proliferation and tryptophanmetabolism.Results:IRF5 was highly expressed in TNBC cell lines.Silencing IRF5 significantly inhibited cellular proliferation and growth.Knockdown of IRF5 reduced the expression of SLC7A5 and IDO1,leading to decreased intracellular levels of tryptophan and its metabolites.ISL markedly suppressed TNBC cell proliferation and disrupted tryptophan metabolism in tumor cells.Conclusion:ISL may inhibit TNBC progression by downregulating IRF5 and interfering with SLC7A5/IDO1-mediated tryptophan metabolic reprogramming,suggesting a potential therapeutic mechanism for TNBC treatment.展开更多
目的采用药效团模型和分子对接方法对ZINC、Chembridge数据库进行虚拟筛选,并通过酶活性测试进行验证,以发现新骨架结构的IDO1抑制剂。方法通过分子对接方法靶向IDO1酶活性位点,对ZINC数据库进行虚拟筛选,得到苗头化合物,进行酶活性测试...目的采用药效团模型和分子对接方法对ZINC、Chembridge数据库进行虚拟筛选,并通过酶活性测试进行验证,以发现新骨架结构的IDO1抑制剂。方法通过分子对接方法靶向IDO1酶活性位点,对ZINC数据库进行虚拟筛选,得到苗头化合物,进行酶活性测试,发现酶活性较好的先导化合物;随后用已进入临床研究的3个IDO1抑制剂构建药效团模型,以此模型对先导化合物类似物进行虚拟筛选,并测定化合物的抑酶活性;通过分子动力学模拟探究化合物与IDO1的结合模式。结果通过分子对接方法对超过200万个虚拟化合物进行筛选得到11个先导化合物并测酶活性,其中ZINC91657208抑酶活性较好,IC50约为77.15μmol/L,活性骨架为4-羟基喹啉。亚结构检索4-羟基喹啉的结构得到31个类似物,利用药效团虚拟筛选出10个化合物,并测酶活性,其中3个4-羟基喹啉类化合物均具有明显的抑酶活性,而Chembridge29374490为酶活性最好的IDO1抑制剂,其IC50约为37.78μmol/L。经分子动力学模拟平衡后,其骨架原子均方差偏根(root mean square deviation, RMSD)分别为1?和2.4?。结论从ZINC和Chembridge数据库中发现了新型4-羟基喹啉类IDO1抑制剂。展开更多
目的调节性T细胞(regulatory T cell,Treg cell)在肿瘤局部免疫耐受中发挥了重要作用,但其产生机制尚不清楚。本实验通过将吲哚胺-2,3-双加氧酶(indoleamine-2,3-dioxygenase,IDO)基因转染小鼠Lewis肺癌细胞,体外研究对Treg细胞的诱导...目的调节性T细胞(regulatory T cell,Treg cell)在肿瘤局部免疫耐受中发挥了重要作用,但其产生机制尚不清楚。本实验通过将吲哚胺-2,3-双加氧酶(indoleamine-2,3-dioxygenase,IDO)基因转染小鼠Lewis肺癌细胞,体外研究对Treg细胞的诱导增殖作用,为肺癌免疫耐受产生机制研究提供新的依据。方法采用阳离子脂质体Lipofectmine2000,将含人全长IDO基因的pEGFP-IDO真核表达质粒转染到小鼠Lewis肺癌细胞,G418筛选获得Lewis-IDO细胞,同时设置空白对照组(Lewis细胞)及空质粒转染组(Lewis-EGFP细胞),将3种细胞分别与小鼠T淋巴细胞混合培养,利用流式细胞技术分析3种细胞对Treg细胞的诱导增殖作用。结果IDO基因成功转染到小鼠Lewis细胞,与小鼠T淋巴细胞共培养后,Treg细胞增殖明显,和Lewis-EG-FP细胞(6.2%:4.9%,P<0.05)及Lewis细胞(6.2%:5.0%,P<0.05)相比差异有统计学意义。结论IDO基因转染小鼠Lew-is肺癌细胞在体外能明显增强Treg细胞的增殖,为肺癌局部免疫耐受机制研究提供了新的实验依据。展开更多
目的探讨吲哚胺-2,3-双加氧酶(indoleamine-2,3-dioxygenase,IDO)基因转染对小鼠Lewis肺癌细胞侵袭和转移能力的影响,并对其机制进行初步探讨。方法通过基因转染技术,利用阳离子脂质体将含有人全长IDO基因序列的真核表达质粒pEGFP-IDO...目的探讨吲哚胺-2,3-双加氧酶(indoleamine-2,3-dioxygenase,IDO)基因转染对小鼠Lewis肺癌细胞侵袭和转移能力的影响,并对其机制进行初步探讨。方法通过基因转染技术,利用阳离子脂质体将含有人全长IDO基因序列的真核表达质粒pEGFP-IDO转染到小鼠Lewis细胞,G418筛选获得Lewis-IDO细胞,设置空白对照组(Lewis细胞)及空质粒转染组(Lewis-EGFP细胞),观察细胞形态学改变,利用transwell侵袭小室检测3种细胞侵袭能力,将3种细胞分别种植到C57/LB小鼠,检测不同细胞种植瘤和转移灶的形成、小鼠外周血中调节性T细胞(Regulatory T cell,Treg)的增殖。结果与Lewis-EGFP细胞和Lewis细胞相比,Lewis-IDO细胞贴壁比例增加,形态变为长梭形,伪足明显增多延长,前二者形态相似;Lewis-IDO细胞侵袭能力增强,破坏matrigel基质蛋白胶层进入下室内的细胞数明显增多,Lewis-IDO细胞、Lewis-EGFP细胞及Lewis细胞分别为237.4±22.1、166.0±14.2和185.6±19.5,前者与后二者比较差异具有统计学意义(P<0.05);种植Lewis-IDO、Lewis-EGFP及Lewis细胞的小鼠发生肝肺转移的小鼠数量及转移灶分别为12只44处、5只7处及4只7处,前者与后二者比较具有统计学意义(P=0.01);流式细胞术分析,种植Lewis-IDO细胞的小鼠外周血中Treg细胞比例明显高于Lewis-EGFP细胞组和Lewis细胞组,其比例分别为(13.7±4.5)%、(8.9±3.3)%和(9.2±3.4)%,前者与后二者比较差异具有统计学意义(P<0.05)。结论IDO基因转染能改变Lewis细胞的培养形态,可以明显增强Lewis细胞的侵袭能力和转移能力,其机制可能与IDO对Lewis细胞形态学改变及诱导Treg细胞的增殖有关。展开更多
文摘妊娠期糖尿病(GDM)是妊娠期间常见的糖代谢异常疾病。本文探讨了吲哚胺2,3-双加氧酶(IDO)与人类白细胞抗原-G (HLA-G)两种免疫调节分子在GDM中的表达及临床意义。本文指明IDO在妊娠期间的活性增加有助于保护胎儿免受母体免疫系统的攻击,但在GDM条件下会造成IDO活性降低。HLA-G通过抑制免疫细胞功能维持母胎间免疫平衡,但GDM会导致血清中可溶性HLA-G水平下降甚至变异。IDO活性降低和HLA-G表达变化提示免疫耐受受损,联合检测这两种分子可为GDM的早期诊断、病情监控及治疗评估提供重要信息。Gestational Diabetes Mellitus (GDM) is a common disorder of glucose metabolism during pregnancy. This article explores the expression and clinical significance of two immunomodulatory molecules, Indoleamine 2,3-Dioxygenase (IDO) and Human Leukocyte Antigen-G (HLA-G), in GDM. It is indicated that the increased activity of IDO during pregnancy helps protect the fetus from maternal immune system attacks, but under GDM conditions, it leads to a reduction in IDO activity. HLA-G maintains immune balance between mother and fetus by inhibiting immune cell function, but GDM causes a decrease or even variation in the levels of soluble HLA-G in the serum. The reduction in IDO activity and changes in HLA-G expression suggest impaired immune tolerance. The combined detection of these two molecules can provide crucial information for the early diagnosis, monitoring, and treatment evaluation of GDM.
基金supported by the Science and Technology Program of Guangzhou(No.202103000020).
文摘Objectives:Triple-negative breast cancer(TNBC)is the breast cancer subtype with the poorest prognosis.This study aimed to elucidate the molecular pathways through which isoliquiritigenin(ISL),a natural chalcone compound derived fromlicorice and other plant roots,targets interferon regulatory factor 5(IRF5)in TNBC.Methods:TNBC cell lines were cultured and subjected to IRF5 knockdown using short hairpin RNA.Cell proliferation was assessed by cell counting kit-8(CCK-8)assay and colony formation assays.Western blotting and quantitative reverse transcription polymerase chain reaction(RT-PCR)were employed to measure expression levels of IRF5,solute carrier family 7 member 5(SLC7A5),and indoleamine 2,3-dioxygenase 1(IDO1).Intracellular tryptophan and its metabolites were quantified using commercially available assay kits and high-performance liquid chromatography(HPLC).TNBC cells were treated with various concentrations of ISL to evaluate its effects on proliferation and tryptophanmetabolism.Results:IRF5 was highly expressed in TNBC cell lines.Silencing IRF5 significantly inhibited cellular proliferation and growth.Knockdown of IRF5 reduced the expression of SLC7A5 and IDO1,leading to decreased intracellular levels of tryptophan and its metabolites.ISL markedly suppressed TNBC cell proliferation and disrupted tryptophan metabolism in tumor cells.Conclusion:ISL may inhibit TNBC progression by downregulating IRF5 and interfering with SLC7A5/IDO1-mediated tryptophan metabolic reprogramming,suggesting a potential therapeutic mechanism for TNBC treatment.
文摘目的采用药效团模型和分子对接方法对ZINC、Chembridge数据库进行虚拟筛选,并通过酶活性测试进行验证,以发现新骨架结构的IDO1抑制剂。方法通过分子对接方法靶向IDO1酶活性位点,对ZINC数据库进行虚拟筛选,得到苗头化合物,进行酶活性测试,发现酶活性较好的先导化合物;随后用已进入临床研究的3个IDO1抑制剂构建药效团模型,以此模型对先导化合物类似物进行虚拟筛选,并测定化合物的抑酶活性;通过分子动力学模拟探究化合物与IDO1的结合模式。结果通过分子对接方法对超过200万个虚拟化合物进行筛选得到11个先导化合物并测酶活性,其中ZINC91657208抑酶活性较好,IC50约为77.15μmol/L,活性骨架为4-羟基喹啉。亚结构检索4-羟基喹啉的结构得到31个类似物,利用药效团虚拟筛选出10个化合物,并测酶活性,其中3个4-羟基喹啉类化合物均具有明显的抑酶活性,而Chembridge29374490为酶活性最好的IDO1抑制剂,其IC50约为37.78μmol/L。经分子动力学模拟平衡后,其骨架原子均方差偏根(root mean square deviation, RMSD)分别为1?和2.4?。结论从ZINC和Chembridge数据库中发现了新型4-羟基喹啉类IDO1抑制剂。
文摘目的调节性T细胞(regulatory T cell,Treg cell)在肿瘤局部免疫耐受中发挥了重要作用,但其产生机制尚不清楚。本实验通过将吲哚胺-2,3-双加氧酶(indoleamine-2,3-dioxygenase,IDO)基因转染小鼠Lewis肺癌细胞,体外研究对Treg细胞的诱导增殖作用,为肺癌免疫耐受产生机制研究提供新的依据。方法采用阳离子脂质体Lipofectmine2000,将含人全长IDO基因的pEGFP-IDO真核表达质粒转染到小鼠Lewis肺癌细胞,G418筛选获得Lewis-IDO细胞,同时设置空白对照组(Lewis细胞)及空质粒转染组(Lewis-EGFP细胞),将3种细胞分别与小鼠T淋巴细胞混合培养,利用流式细胞技术分析3种细胞对Treg细胞的诱导增殖作用。结果IDO基因成功转染到小鼠Lewis细胞,与小鼠T淋巴细胞共培养后,Treg细胞增殖明显,和Lewis-EG-FP细胞(6.2%:4.9%,P<0.05)及Lewis细胞(6.2%:5.0%,P<0.05)相比差异有统计学意义。结论IDO基因转染小鼠Lew-is肺癌细胞在体外能明显增强Treg细胞的增殖,为肺癌局部免疫耐受机制研究提供了新的实验依据。
文摘目的探讨吲哚胺-2,3-双加氧酶(indoleamine-2,3-dioxygenase,IDO)基因转染对小鼠Lewis肺癌细胞侵袭和转移能力的影响,并对其机制进行初步探讨。方法通过基因转染技术,利用阳离子脂质体将含有人全长IDO基因序列的真核表达质粒pEGFP-IDO转染到小鼠Lewis细胞,G418筛选获得Lewis-IDO细胞,设置空白对照组(Lewis细胞)及空质粒转染组(Lewis-EGFP细胞),观察细胞形态学改变,利用transwell侵袭小室检测3种细胞侵袭能力,将3种细胞分别种植到C57/LB小鼠,检测不同细胞种植瘤和转移灶的形成、小鼠外周血中调节性T细胞(Regulatory T cell,Treg)的增殖。结果与Lewis-EGFP细胞和Lewis细胞相比,Lewis-IDO细胞贴壁比例增加,形态变为长梭形,伪足明显增多延长,前二者形态相似;Lewis-IDO细胞侵袭能力增强,破坏matrigel基质蛋白胶层进入下室内的细胞数明显增多,Lewis-IDO细胞、Lewis-EGFP细胞及Lewis细胞分别为237.4±22.1、166.0±14.2和185.6±19.5,前者与后二者比较差异具有统计学意义(P<0.05);种植Lewis-IDO、Lewis-EGFP及Lewis细胞的小鼠发生肝肺转移的小鼠数量及转移灶分别为12只44处、5只7处及4只7处,前者与后二者比较具有统计学意义(P=0.01);流式细胞术分析,种植Lewis-IDO细胞的小鼠外周血中Treg细胞比例明显高于Lewis-EGFP细胞组和Lewis细胞组,其比例分别为(13.7±4.5)%、(8.9±3.3)%和(9.2±3.4)%,前者与后二者比较差异具有统计学意义(P<0.05)。结论IDO基因转染能改变Lewis细胞的培养形态,可以明显增强Lewis细胞的侵袭能力和转移能力,其机制可能与IDO对Lewis细胞形态学改变及诱导Treg细胞的增殖有关。
