高迁移率族蛋白1(high mobility group box 1,HMGB1)是一种高度保守的DNA结合蛋白,与核小体结构的修复及基因转录调控密切相关。HMGB1由坏死细胞或活化的单核/巨噬细胞被动或主动释放至细胞外,并发挥促炎和修复作用。脓毒症是一种全身...高迁移率族蛋白1(high mobility group box 1,HMGB1)是一种高度保守的DNA结合蛋白,与核小体结构的修复及基因转录调控密切相关。HMGB1由坏死细胞或活化的单核/巨噬细胞被动或主动释放至细胞外,并发挥促炎和修复作用。脓毒症是一种全身炎症反应综合征。发生脓毒症时,血清和组织中HMGB1浓度上升,HMGB1在其发生过程中扮演晚期促炎细胞因子的角色。在脓毒症和内毒素血症动物模型中,靶向给予HMGB1抗体或特异性拮抗剂可以改善预后。本文就HMGB1的结构、在细胞核内发挥的作用、释放过程和其在炎症反应及脓毒症中发挥的作用研究进展作一综述。展开更多
Objective： To detect the expression profiles of micro RNA-218（mi R-218） in human pancreatic cancer tissue（PCT） and cells and their effects on the biological features of human pancreatic cancer cell line PANC-1 an...Objective： To detect the expression profiles of micro RNA-218（mi R-218） in human pancreatic cancer tissue（PCT） and cells and their effects on the biological features of human pancreatic cancer cell line PANC-1 and observe the effect of mi R-218 on the expression of the target gene high mobility group box 1（HMGB1）, with an attempt to provide new treatment methods and strategies for pancreatic cancer.Methods： The expressions of mi R-218 in PCT and normal pancreas tissue as well as in various pancreatic cancer cell lines including As PC-1, Bx PC-3, and PANC-1 were determined with quantitative real-time reverse transcription polymerase chain reaction（q RT-PCR）. The change of mi R-218 expression in PANC-1 cells was detected using qR T-PCT after the transfection of miR-218 mimic for 48 h. Cell Counting Kit-8（CCK-8） was applied for detecting the effect of mi R-218 on the activity of PANC-1 cells. The effects of mi R-218 on the proliferation and apoptosis of PANC-1 cells were analyzed using the flow cytometry. The effect of mi R-218 on the migration of PANC-1 cells was detected using the Trans-well migration assay. The HMGB1 was found to be a target gene of mi R-218 by luciferase reporter assay, and the effect of mi R-218 on the expression of HMGB1 protein in cells were determined using Western blotting.Results： As shown by q RT-PCR, the expressions of mi R-218 in PCT and in pancreatic cancer cell line significantly decreased when compared with the normal pancreatic tissue（NPT）（P〈0.01）. Compared with the control group, the miR-218 expression significantly increased in the PANC-1 group after the transfection of mi R-218 mimic for 48 h（P〈0.01）. Growth curve showed that the cell viability significantly dropped after the overexpression of mi R-218 in the PANC-1 cells for two days（P〈0.05）. Flow cytometry showed that the S-phase fraction significantly dropped after the overexpression of mi R-218（P〈0.01） and the percentage of apoptotic cells significantly increased（P〈0.0展开更多
目的:探讨miR-129-5p通过调控高迁移率族蛋白B1基因(high mobility group box 1,HMGB1)影响乳腺癌MCF-7细胞对紫杉醇(paclitaxel,PTX)的敏感性。方法:采用脂质体转染技术将miR-129-5p mimics、HMGB1小干扰RNA(si-HMGB1)分别转染入MCF-7...目的:探讨miR-129-5p通过调控高迁移率族蛋白B1基因(high mobility group box 1,HMGB1)影响乳腺癌MCF-7细胞对紫杉醇(paclitaxel,PTX)的敏感性。方法:采用脂质体转染技术将miR-129-5p mimics、HMGB1小干扰RNA(si-HMGB1)分别转染入MCF-7细胞,用PTX刺激培养细胞后,用实时荧光定量PCR检测转染后MCF-7细胞miR-129-5p和HMGB1 m RNA的表达,Western blotting检测转染后MCF-7细胞HMGB1蛋白的表达,CCK-8增殖实验检测转染后PTX对MCF-7细胞增殖的影响,流式细胞术检测转染后对PTX诱导MCF-7细胞凋亡的影响。结果:转染miR-129-5p mimics后,MCF-7细胞中miR-129-5p的表达水平明显高于阴性对照组细胞(P<0.01);过表达miR-129-5p后可明显增强PTX抑制MCF-7细胞的增殖和诱导细胞凋亡的能力(均P<0.05),并显著抑制HMGB1 m RNA和蛋白的表达(均P<0.05)。转染si-HMGB1后,显著降低MCF-7细胞HMGB1 m RNA和蛋白的表达(均P<0.05);干扰HMGB1表达进一步促进PTX抑制MCF-7细胞的增殖并诱导细胞凋亡(均P<0.05)。结论:miR-129-5p通过下调HMGB1的表达增强乳腺癌MCF-7细胞对PTX的敏感性。展开更多
Gram-negative sepsis is a severe clinical syndrome associated with significant morbidity and mortality.Lipopolysaccharide(LPS),expressed on Gram-negative bacteria,is a potent pro-inflammatory toxin that induces inflam...Gram-negative sepsis is a severe clinical syndrome associated with significant morbidity and mortality.Lipopolysaccharide(LPS),expressed on Gram-negative bacteria,is a potent pro-inflammatory toxin that induces inflammation and coagulation via two separate receptor systems.One is Toll-like receptor 4(TLR4),expressed on cell surfaces and in endosomes,and the other is the cytosolic receptor caspase-11(caspases-4 and-5 in hu-mans).Extracellular LPS binds to high mobility group box 1(HMGB1)protein,a cytokine-like molecule.The HMGB1-LPS complex is transported via receptor for advanced glycated end products(RAGE)-endocytosis to the endolysosomal system to reach the cytosolic LPS receptor caspase-11 to induce HMGB1 release,inflammation,and coagulation that may cause multi-organ failure.The insight that LPS needs HMGB1 assistance to generate severe inflammation has led to successful therapeutic results in preclinical Gram-negative sepsis studies target-ing HMGB1.However,to date,no clinical studies have been performed based on this strategy.HMGB1 is also actively released by peripheral sensory nerves and this mechanism is fundamental for the initiation and prop-agation of inflammation during tissue injury.Homeostasis is achieved when other neurons actively restrict the inflammatory response via monitoring by the central nervous system and the vagus nerve through the cholinergic anti-inflammatory pathway.The neuronal control in Gram-negative sepsis needs further studies since a deeper understanding of the interplay between HMGB1 and acetylcholine may have beneficial therapeutic implications.Herein,we review the synergistic overlapping mechanisms of LPS and HMGB1 and discuss future treatment opportunities in Gram-negative sepsis.展开更多
Objective Innate lymphoid cells(ILCs)are a class of newly discovered immunocytes.Group 1 ILCs(ILC1s)are identified in the decidua of humans and mice.High mobility group box 1(HMGB1)is predicted to be one of the target...Objective Innate lymphoid cells(ILCs)are a class of newly discovered immunocytes.Group 1 ILCs(ILC1s)are identified in the decidua of humans and mice.High mobility group box 1(HMGB1)is predicted to be one of the target genes of miR-142-3p,which is closely related to pregnancy-related diseases.Furthermore,miR-142-3p and HMGB1 are involved in regulating the NF-κB signaling pathway.This study aimed to examine the regulatory effect of miR-142-3p on ILC1s and the underlying mechanism involving HMGB1 and the NF-κB signaling pathway.Methods Mouse models of normal pregnancy and abortion were constructed,and the alterations of ILC1s,miR-142-3p,ILC1 transcription factor(T-bet),and pro-inflammatory cytokines of ILC1s(TNF-α,IFN-γand IL-2)were detected in mice from different groups.The targeting regulation of HMGB1 by miR-142-3p in ILC1s,and the expression of HMGB1 in normal pregnant mice and abortive mice were investigated.In addition,the regulatory effects of miR-142-3p and HMGB1 on ILC1s were detected in vitro by CCK-8,Annexin-V/PI,ELISA,and RT-PCR,respectively.Furthermore,changes of the NF-κB signaling pathway in ILC1s were examined in the different groups.For the in vivo studies,miR-142-3p-Agomir was injected in the uterus of abortive mice to evaluate the abortion rate and alterations of ILC1s at the maternal-fetal interface,and further detect the expression of HMGB1,pro-inflammatory cytokines,and the NF-κB signaling pathway.Results The number of ILC1s was significantly increased,the level of HMGB1 was significantly upregulated,and that of miR-142-3p was considerably downregulated in the abortive mice as compared with the normal pregnant mice(all P<0.05).In addition,miR-142-3p was found to drastically inhibit the activation of the NF-κB signaling pathway(P<0.05).The number of ILC1s and the levels of pro-inflammatory cytokines were significantly downregulated and the activation of the NF-κB signaling pathway was inhibited in the miR-142-3p Agomir group(all P<0.05).Conclusion miR-142-3p can regulate ILC1s by targeti展开更多
文摘高迁移率族蛋白1(high mobility group box 1,HMGB1)是一种高度保守的DNA结合蛋白,与核小体结构的修复及基因转录调控密切相关。HMGB1由坏死细胞或活化的单核/巨噬细胞被动或主动释放至细胞外,并发挥促炎和修复作用。脓毒症是一种全身炎症反应综合征。发生脓毒症时,血清和组织中HMGB1浓度上升,HMGB1在其发生过程中扮演晚期促炎细胞因子的角色。在脓毒症和内毒素血症动物模型中,靶向给予HMGB1抗体或特异性拮抗剂可以改善预后。本文就HMGB1的结构、在细胞核内发挥的作用、释放过程和其在炎症反应及脓毒症中发挥的作用研究进展作一综述。
基金supported by grants:Liaoning Provincial Department of education science research project(L2014299)Liaoning province science and technology plan project(2011404013-4)The Shenyang Municipal Science and technology project(F12-277-1-73)
文摘Objective： To detect the expression profiles of micro RNA-218（mi R-218） in human pancreatic cancer tissue（PCT） and cells and their effects on the biological features of human pancreatic cancer cell line PANC-1 and observe the effect of mi R-218 on the expression of the target gene high mobility group box 1（HMGB1）, with an attempt to provide new treatment methods and strategies for pancreatic cancer.Methods： The expressions of mi R-218 in PCT and normal pancreas tissue as well as in various pancreatic cancer cell lines including As PC-1, Bx PC-3, and PANC-1 were determined with quantitative real-time reverse transcription polymerase chain reaction（q RT-PCR）. The change of mi R-218 expression in PANC-1 cells was detected using qR T-PCT after the transfection of miR-218 mimic for 48 h. Cell Counting Kit-8（CCK-8） was applied for detecting the effect of mi R-218 on the activity of PANC-1 cells. The effects of mi R-218 on the proliferation and apoptosis of PANC-1 cells were analyzed using the flow cytometry. The effect of mi R-218 on the migration of PANC-1 cells was detected using the Trans-well migration assay. The HMGB1 was found to be a target gene of mi R-218 by luciferase reporter assay, and the effect of mi R-218 on the expression of HMGB1 protein in cells were determined using Western blotting.Results： As shown by q RT-PCR, the expressions of mi R-218 in PCT and in pancreatic cancer cell line significantly decreased when compared with the normal pancreatic tissue（NPT）（P〈0.01）. Compared with the control group, the miR-218 expression significantly increased in the PANC-1 group after the transfection of mi R-218 mimic for 48 h（P〈0.01）. Growth curve showed that the cell viability significantly dropped after the overexpression of mi R-218 in the PANC-1 cells for two days（P〈0.05）. Flow cytometry showed that the S-phase fraction significantly dropped after the overexpression of mi R-218（P〈0.01） and the percentage of apoptotic cells significantly increased（P〈0.0
文摘目的:探讨miR-129-5p通过调控高迁移率族蛋白B1基因(high mobility group box 1,HMGB1)影响乳腺癌MCF-7细胞对紫杉醇(paclitaxel,PTX)的敏感性。方法:采用脂质体转染技术将miR-129-5p mimics、HMGB1小干扰RNA(si-HMGB1)分别转染入MCF-7细胞,用PTX刺激培养细胞后,用实时荧光定量PCR检测转染后MCF-7细胞miR-129-5p和HMGB1 m RNA的表达,Western blotting检测转染后MCF-7细胞HMGB1蛋白的表达,CCK-8增殖实验检测转染后PTX对MCF-7细胞增殖的影响,流式细胞术检测转染后对PTX诱导MCF-7细胞凋亡的影响。结果:转染miR-129-5p mimics后,MCF-7细胞中miR-129-5p的表达水平明显高于阴性对照组细胞(P<0.01);过表达miR-129-5p后可明显增强PTX抑制MCF-7细胞的增殖和诱导细胞凋亡的能力(均P<0.05),并显著抑制HMGB1 m RNA和蛋白的表达(均P<0.05)。转染si-HMGB1后,显著降低MCF-7细胞HMGB1 m RNA和蛋白的表达(均P<0.05);干扰HMGB1表达进一步促进PTX抑制MCF-7细胞的增殖并诱导细胞凋亡(均P<0.05)。结论:miR-129-5p通过下调HMGB1的表达增强乳腺癌MCF-7细胞对PTX的敏感性。
文摘Gram-negative sepsis is a severe clinical syndrome associated with significant morbidity and mortality.Lipopolysaccharide(LPS),expressed on Gram-negative bacteria,is a potent pro-inflammatory toxin that induces inflammation and coagulation via two separate receptor systems.One is Toll-like receptor 4(TLR4),expressed on cell surfaces and in endosomes,and the other is the cytosolic receptor caspase-11(caspases-4 and-5 in hu-mans).Extracellular LPS binds to high mobility group box 1(HMGB1)protein,a cytokine-like molecule.The HMGB1-LPS complex is transported via receptor for advanced glycated end products(RAGE)-endocytosis to the endolysosomal system to reach the cytosolic LPS receptor caspase-11 to induce HMGB1 release,inflammation,and coagulation that may cause multi-organ failure.The insight that LPS needs HMGB1 assistance to generate severe inflammation has led to successful therapeutic results in preclinical Gram-negative sepsis studies target-ing HMGB1.However,to date,no clinical studies have been performed based on this strategy.HMGB1 is also actively released by peripheral sensory nerves and this mechanism is fundamental for the initiation and prop-agation of inflammation during tissue injury.Homeostasis is achieved when other neurons actively restrict the inflammatory response via monitoring by the central nervous system and the vagus nerve through the cholinergic anti-inflammatory pathway.The neuronal control in Gram-negative sepsis needs further studies since a deeper understanding of the interplay between HMGB1 and acetylcholine may have beneficial therapeutic implications.Herein,we review the synergistic overlapping mechanisms of LPS and HMGB1 and discuss future treatment opportunities in Gram-negative sepsis.
基金supported by the National Key Research and Development Program of China(Nos.2018YFC1002804 and 2016YFC1000600)the National Natural Science Foundation of China(Nos.81771618 and 81971356)the Fundamental Research Funds for the Central Universities(No.2042023kf0028).
文摘Objective Innate lymphoid cells(ILCs)are a class of newly discovered immunocytes.Group 1 ILCs(ILC1s)are identified in the decidua of humans and mice.High mobility group box 1(HMGB1)is predicted to be one of the target genes of miR-142-3p,which is closely related to pregnancy-related diseases.Furthermore,miR-142-3p and HMGB1 are involved in regulating the NF-κB signaling pathway.This study aimed to examine the regulatory effect of miR-142-3p on ILC1s and the underlying mechanism involving HMGB1 and the NF-κB signaling pathway.Methods Mouse models of normal pregnancy and abortion were constructed,and the alterations of ILC1s,miR-142-3p,ILC1 transcription factor(T-bet),and pro-inflammatory cytokines of ILC1s(TNF-α,IFN-γand IL-2)were detected in mice from different groups.The targeting regulation of HMGB1 by miR-142-3p in ILC1s,and the expression of HMGB1 in normal pregnant mice and abortive mice were investigated.In addition,the regulatory effects of miR-142-3p and HMGB1 on ILC1s were detected in vitro by CCK-8,Annexin-V/PI,ELISA,and RT-PCR,respectively.Furthermore,changes of the NF-κB signaling pathway in ILC1s were examined in the different groups.For the in vivo studies,miR-142-3p-Agomir was injected in the uterus of abortive mice to evaluate the abortion rate and alterations of ILC1s at the maternal-fetal interface,and further detect the expression of HMGB1,pro-inflammatory cytokines,and the NF-κB signaling pathway.Results The number of ILC1s was significantly increased,the level of HMGB1 was significantly upregulated,and that of miR-142-3p was considerably downregulated in the abortive mice as compared with the normal pregnant mice(all P<0.05).In addition,miR-142-3p was found to drastically inhibit the activation of the NF-κB signaling pathway(P<0.05).The number of ILC1s and the levels of pro-inflammatory cytokines were significantly downregulated and the activation of the NF-κB signaling pathway was inhibited in the miR-142-3p Agomir group(all P<0.05).Conclusion miR-142-3p can regulate ILC1s by targeti
