Background:Cataracts are the leading cause of reversible blindness worldwide.Diabetic cataract(DC),a prevalent complication of diabetes mellitus,is characterized by its high occurrence,rapid progression,and severe imp...Background:Cataracts are the leading cause of reversible blindness worldwide.Diabetic cataract(DC),a prevalent complication of diabetes mellitus,is characterized by its high occurrence,rapid progression,and severe impact.The prevalence of diabetes varies greatly between the northern and southern regions,with higher rates observed among northern residents.DC-induced lens opacity is mainly attributed to oxidative stress.However,it remains unclear whether ferroptosis,a form of regulated cell death,occurs in crystalline epithelial cells during the pathogenesis,which may represent a novel mechanism contributing to DC.Methods:Transmission electron microscopy,quantitative assays for iron levels and reactive oxygen species(ROS),real-time quantitative polymerase chain reaction(RT-qPCR),western blotting,immunofluorescence,and immunohistochemistry were used to detect ferroptosis.Gene editing techniques were utilized to study the regulatory relationships among lipocalin 2(LCN2),glutathione peroxidase 4(GPX4),and ferritin heavy chain(FTH).Local knockdown of the LCN2 gene in B-3 cells and the eyes of Sprague Dawley(SD)rats was performed to verify and further explore the role and regulatory mechanisms of LCN2 in DC-associated ferroptosis.Results:An in vitro model using high glucose levels and an in vivo model with streptozotocin-induced diabetes in SD rats were successfully established.Ferroptosis was observed in both in vitro and in vivo experiments.LCN2 protein was normally expressed in human and rat lens epithelial cells,but its expression significantly increased during ferroptosis.The ferroptosis inhibitor,ferrostatin-1(Fer-1)effectively inhibited ferroptosis and reduced LCN2 protein expression.Notably,local knockdown of LCN2 via gene editing protected lens epithelial cells from ferroptosis in vitro and slowed the progression of DC in SD rats in vivo.Conclusion:Our findings underscore the significant role of ferroptosis in the pathogenesis of DC,suggesting that selectively targeting LCN2 activation and enhancing ferroptosis resistance may offer a novel therapeutic approach for treating DC.展开更多
Diabetes is a major global public health issue.The prevalence of type 1 diabetes is comparatively static,as hereditary and genetic causes are involved,while type 2 diabetes(T2D)prevalence is increasing day by day.T2D ...Diabetes is a major global public health issue.The prevalence of type 1 diabetes is comparatively static,as hereditary and genetic causes are involved,while type 2 diabetes(T2D)prevalence is increasing day by day.T2D is associated with chronic complications,including diabetic neuropathy(DN),nephropathy,retinopathy,and other complications like diabetic foot.DN is the main complication of both types of diabetes.DN can be diagnosed by routine laboratory tests,microalbuminuria>300 mg/24 h,and a gradual decrease in glomerular filtration rate.As the appearance of microalbuminuria is a late manifestation,an early marker for renal damage is needed.Lipocalin-2,also known as neutrophil gelatinaseassociated lipocalin(NGAL),is a small protein purified from neutrophil granules and a good marker for kidney disease.NGAL is a transporter protein responsible for many physiological processes,such as inflammation,generation of the immune response,and metabolic homeostasis.NGAL has been reported to depict the early changes in renal damage when urine microalbumin is still undetecable.Therefore,elucidating the role of NGAL in detecting DN and understanding its mechanism can help establish it as a potential early marker for DN.展开更多
Objective To explore whether the amount of lipocalin-2 in the biofluid could reflect the onset of sepsis-induced acute lung injury(ALI) in mice. Methods Lipopolysaccharide(LPS, 10 mg/kg) injection or cecal ligation an...Objective To explore whether the amount of lipocalin-2 in the biofluid could reflect the onset of sepsis-induced acute lung injury(ALI) in mice. Methods Lipopolysaccharide(LPS, 10 mg/kg) injection or cecal ligation and puncture(CLP) was performed to induce severe sepsis and ALI in C57 BL/6 male mice randomly divided into 5 groups(n=10 in each group): group A(intraperitoneal LPS injection), group B(intravenous LPS injection via tail vein), group C(CLP with 25% of the cecum ligated), group D(CLP with 75% of the cecum ligated), and the control group(6 sham-operation controls plus 4 saline controls). All the mice received volume resuscitation. Measurements of pulmonary morphological and functional alterations were used to identify the presence of experimental ALI. The expressions of lipocalin-2 and interleukin(IL)-6 in serum, bronchoalveolar lavage fluid(BALF), and lung tissue were quantified at both protein and mRNA levels. The overall abilities of lipocalin-2 and IL-6 tests to diagnose sepsis-induced ALI were evaluated by generating receiver operator characteristic curves(ROC) and computing area under curve(AUC). Results In both group B and group D, most of the "main features" of experimental ALI were reproduced in mice, while group A and group C showed septic syndrome without definite evidence for the presence of ALI. Compared with septic mice without ALI(group A+group C), lipocalin-2 protein expression in septic mice with ALI(group B+group D) was significantly up-regulated in BALF(P<0.01) and in serum(P<0.01), and mRNA expression boosted in lung tissues(all P<0.05). Lipocalin-2 tests performed better than IL-6 tests in recognizing sepsis-induced ALI cases, evidenced by the larger AUC of the former(BALF tests, 0.8800 versus 0.6625; serum tests, 0.8500 versus 0.7000). Using a dual cutoff system to diagnose sepsis-induced ALI, BALF lipocalin-2 test exhibited the highest positive likelihood ratio(13.000) and the lowest negative likelihood ratio(0.077) among the tests of lipocalin-2 and IL-6 in blood and BALF. A statistically significant correlation was found between lipocalin-2 concentration in BALF and that in serum(Spearman r=0.8803,P<0.0001). Conclusions Lipocalin-2 expression is significantly up-regulated in septic ALI mice compared with those without ALI. Lipocalin-2 tests with a dual cutoff system could be an effective tool in distinguishing experimental ALI cases.展开更多
Purpose of the study: Research of the clinical and diagnostic significance of determination of Lipocalin-2 associated with neutrophilic gelatinases (uNGAL) in the urine of children with urinary tract infection (UTI) a...Purpose of the study: Research of the clinical and diagnostic significance of determination of Lipocalin-2 associated with neutrophilic gelatinases (uNGAL) in the urine of children with urinary tract infection (UTI) and pyelonephritis. Materials and methods: We examined 30 children with acute pyelonephritis and UTI aged 1 to 16 years (average age 7.32 ± 4.52) including 26 girls and 4 boys. Verification of the diagnosis was conducted on the basis of clinical and laboratory data, medical history and instrumental examination of patients. All children were divided into 2 groups: 1st group—15 children with acute pyelonephritis, 2nd group—15 children with urinary tract infection. uNGAL was measured in the urine by enzyme-linked immunosorbent assay (EISA) (BioVendor Laboratoty Medicine). Results: It is found, that the urine level of NGAL depends on the damage degree of renal parenchyma. The correlation of medium strength was found between the excretion level of uNGAL during the acute period of pyelonephritis and the detection of renal scars according to the DMSA-nephroscintigraphy data. In the group of children with the acute pyelonephritis the direct correlation of medium strength was found between the excretion level of uNGAL/creatinine and leukocytosis value and also with the CRP blood level. Conclusion: The results allow us to recommend the determination of the excretion level of uNGAL/creatinine as an additional non-invasive marker for the early detection of renal parenchyma injury.展开更多
Hepatic stellate cells(HSCs)represent a significant component of hepatocellular carcinoma(HCC)microenvironments which play a critical role in tumor progression and drug resistance.Tumor-ona-chip technology has provide...Hepatic stellate cells(HSCs)represent a significant component of hepatocellular carcinoma(HCC)microenvironments which play a critical role in tumor progression and drug resistance.Tumor-ona-chip technology has provided a powerful in vitro platform to investigate the crosstalk between activated HSCs and HCC cells by mimicking physiological architecture with precise spatiotemporal control.Here we developed a tri-cell culture microfluidic chip to evaluate the impact of HSCs on HCC progression.Onchip analysis revealed activated HSCs contributed to endothelial invasion,HCC drug resistance and natural killer(NK)cell exhaustion.Cytokine array and RNA sequencing analysis were combined to indicate the iron-binding protein LIPOCALIN-2(LCN-2)as a key factor in remodeling tumor microenvironments in the HCC-on-a-chip.LCN-2 targeted therapy demonstrated robust anti-tumor effects both in vitro 3D biomimetic chip and in vivo mouse model,including angiogenesis inhibition,sorafenib sensitivity promotion and NK-cell cytotoxicity enhancement.Taken together,the microfluidic platform exhibited obvious advantages in mimicking functional characteristics of tumor microenvironments and developing targeted therapies.展开更多
Clinical cases have reported pulmonary arterial structural and functional abnormalities in patients with Kawasaki disease(KD);however,the underlying mechanisms are unclear.In this study,a KD rat model was established ...Clinical cases have reported pulmonary arterial structural and functional abnormalities in patients with Kawasaki disease(KD);however,the underlying mechanisms are unclear.In this study,a KD rat model was established via the intraperitoneal injection of Lactobacillus casei cell wall extract(LCWE).The results showed that pulmonary arterial functional and structural abnormalities were observed in KD rats.Furthermore,proliferative endoplasmic reticulum stress(ER stress)was observed in the pulmonary arteries of KD rats.Notably,the level of lipocalin-2(Lcn 2),a trigger factor of inflammation,was remarkably elevated in the plasma and lung tissues of KD rats;increased Lcn 2 levels following LCWE stimulation may result from polymorphonuclear neutrophils(PMNs).Correspondingly,in cultured pulmonary artery smooth muscle cells(PASMCs),Lcn 2 markedly augmented the cleavage and nuclear localization of activating transcription factor-6(ATF6),upregulated the transcription of glucose regulated protein 78(GRP78)and neurite outgrowth inhibitor(NOGO),and promoted PASMCs proliferation.However,proapoptotic C/EBP homologous protein(CHOP)and caspase 12 levels were not elevated.Treatment with 4-phenyl butyric acid(4-PBA,a specific inhibitor of ER stress)inhibited PASMCs proliferation induced by Lcn 2 and attenuated pulmonary arterial abnormalities and right ventricular hypertrophy and reduced right ventricular systolic pressure in KD rats.In conclusion,Lcn 2 remarkably facilitates proliferative ER stress in PASMCs,which probably accounts for KD-related pulmonary arterial abnormalities.展开更多
基金approved by the Institutional Review Board(or Ethics Committee)of the First Affiliated Hospital of Harbin Medical University(IACUC:2022020 and date of approval:April 8,2022).
文摘Background:Cataracts are the leading cause of reversible blindness worldwide.Diabetic cataract(DC),a prevalent complication of diabetes mellitus,is characterized by its high occurrence,rapid progression,and severe impact.The prevalence of diabetes varies greatly between the northern and southern regions,with higher rates observed among northern residents.DC-induced lens opacity is mainly attributed to oxidative stress.However,it remains unclear whether ferroptosis,a form of regulated cell death,occurs in crystalline epithelial cells during the pathogenesis,which may represent a novel mechanism contributing to DC.Methods:Transmission electron microscopy,quantitative assays for iron levels and reactive oxygen species(ROS),real-time quantitative polymerase chain reaction(RT-qPCR),western blotting,immunofluorescence,and immunohistochemistry were used to detect ferroptosis.Gene editing techniques were utilized to study the regulatory relationships among lipocalin 2(LCN2),glutathione peroxidase 4(GPX4),and ferritin heavy chain(FTH).Local knockdown of the LCN2 gene in B-3 cells and the eyes of Sprague Dawley(SD)rats was performed to verify and further explore the role and regulatory mechanisms of LCN2 in DC-associated ferroptosis.Results:An in vitro model using high glucose levels and an in vivo model with streptozotocin-induced diabetes in SD rats were successfully established.Ferroptosis was observed in both in vitro and in vivo experiments.LCN2 protein was normally expressed in human and rat lens epithelial cells,but its expression significantly increased during ferroptosis.The ferroptosis inhibitor,ferrostatin-1(Fer-1)effectively inhibited ferroptosis and reduced LCN2 protein expression.Notably,local knockdown of LCN2 via gene editing protected lens epithelial cells from ferroptosis in vitro and slowed the progression of DC in SD rats in vivo.Conclusion:Our findings underscore the significant role of ferroptosis in the pathogenesis of DC,suggesting that selectively targeting LCN2 activation and enhancing ferroptosis resistance may offer a novel therapeutic approach for treating DC.
文摘Diabetes is a major global public health issue.The prevalence of type 1 diabetes is comparatively static,as hereditary and genetic causes are involved,while type 2 diabetes(T2D)prevalence is increasing day by day.T2D is associated with chronic complications,including diabetic neuropathy(DN),nephropathy,retinopathy,and other complications like diabetic foot.DN is the main complication of both types of diabetes.DN can be diagnosed by routine laboratory tests,microalbuminuria>300 mg/24 h,and a gradual decrease in glomerular filtration rate.As the appearance of microalbuminuria is a late manifestation,an early marker for renal damage is needed.Lipocalin-2,also known as neutrophil gelatinaseassociated lipocalin(NGAL),is a small protein purified from neutrophil granules and a good marker for kidney disease.NGAL is a transporter protein responsible for many physiological processes,such as inflammation,generation of the immune response,and metabolic homeostasis.NGAL has been reported to depict the early changes in renal damage when urine microalbumin is still undetecable.Therefore,elucidating the role of NGAL in detecting DN and understanding its mechanism can help establish it as a potential early marker for DN.
基金Supported in part by Jie-shou Li Academician Gut Barrier Research Fund(2012001)
文摘Objective To explore whether the amount of lipocalin-2 in the biofluid could reflect the onset of sepsis-induced acute lung injury(ALI) in mice. Methods Lipopolysaccharide(LPS, 10 mg/kg) injection or cecal ligation and puncture(CLP) was performed to induce severe sepsis and ALI in C57 BL/6 male mice randomly divided into 5 groups(n=10 in each group): group A(intraperitoneal LPS injection), group B(intravenous LPS injection via tail vein), group C(CLP with 25% of the cecum ligated), group D(CLP with 75% of the cecum ligated), and the control group(6 sham-operation controls plus 4 saline controls). All the mice received volume resuscitation. Measurements of pulmonary morphological and functional alterations were used to identify the presence of experimental ALI. The expressions of lipocalin-2 and interleukin(IL)-6 in serum, bronchoalveolar lavage fluid(BALF), and lung tissue were quantified at both protein and mRNA levels. The overall abilities of lipocalin-2 and IL-6 tests to diagnose sepsis-induced ALI were evaluated by generating receiver operator characteristic curves(ROC) and computing area under curve(AUC). Results In both group B and group D, most of the "main features" of experimental ALI were reproduced in mice, while group A and group C showed septic syndrome without definite evidence for the presence of ALI. Compared with septic mice without ALI(group A+group C), lipocalin-2 protein expression in septic mice with ALI(group B+group D) was significantly up-regulated in BALF(P<0.01) and in serum(P<0.01), and mRNA expression boosted in lung tissues(all P<0.05). Lipocalin-2 tests performed better than IL-6 tests in recognizing sepsis-induced ALI cases, evidenced by the larger AUC of the former(BALF tests, 0.8800 versus 0.6625; serum tests, 0.8500 versus 0.7000). Using a dual cutoff system to diagnose sepsis-induced ALI, BALF lipocalin-2 test exhibited the highest positive likelihood ratio(13.000) and the lowest negative likelihood ratio(0.077) among the tests of lipocalin-2 and IL-6 in blood and BALF. A statistically significant correlation was found between lipocalin-2 concentration in BALF and that in serum(Spearman r=0.8803,P<0.0001). Conclusions Lipocalin-2 expression is significantly up-regulated in septic ALI mice compared with those without ALI. Lipocalin-2 tests with a dual cutoff system could be an effective tool in distinguishing experimental ALI cases.
文摘Purpose of the study: Research of the clinical and diagnostic significance of determination of Lipocalin-2 associated with neutrophilic gelatinases (uNGAL) in the urine of children with urinary tract infection (UTI) and pyelonephritis. Materials and methods: We examined 30 children with acute pyelonephritis and UTI aged 1 to 16 years (average age 7.32 ± 4.52) including 26 girls and 4 boys. Verification of the diagnosis was conducted on the basis of clinical and laboratory data, medical history and instrumental examination of patients. All children were divided into 2 groups: 1st group—15 children with acute pyelonephritis, 2nd group—15 children with urinary tract infection. uNGAL was measured in the urine by enzyme-linked immunosorbent assay (EISA) (BioVendor Laboratoty Medicine). Results: It is found, that the urine level of NGAL depends on the damage degree of renal parenchyma. The correlation of medium strength was found between the excretion level of uNGAL during the acute period of pyelonephritis and the detection of renal scars according to the DMSA-nephroscintigraphy data. In the group of children with the acute pyelonephritis the direct correlation of medium strength was found between the excretion level of uNGAL/creatinine and leukocytosis value and also with the CRP blood level. Conclusion: The results allow us to recommend the determination of the excretion level of uNGAL/creatinine as an additional non-invasive marker for the early detection of renal parenchyma injury.
基金funded by National Natural Science Foundation of China (Nos.31901010 and 81803916,China)Natural Science Foundation of Jiangsu Province (grant No.BK20180128,China)+3 种基金The Priority Academic Program Development of Jiangsu Higher Education Institutions (Integration of Chinese and Western Medicine,China)Jiangsu Specially Appointed ProfessorshipJiangsu Key Discipline Construction Fund of the 14th Five-Year Plan (Biology,China)The Graduate Research&Practice Innovation Program of Jiangsu Province (KYCX21_1743,KYCX22_1992,China)。
文摘Hepatic stellate cells(HSCs)represent a significant component of hepatocellular carcinoma(HCC)microenvironments which play a critical role in tumor progression and drug resistance.Tumor-ona-chip technology has provided a powerful in vitro platform to investigate the crosstalk between activated HSCs and HCC cells by mimicking physiological architecture with precise spatiotemporal control.Here we developed a tri-cell culture microfluidic chip to evaluate the impact of HSCs on HCC progression.Onchip analysis revealed activated HSCs contributed to endothelial invasion,HCC drug resistance and natural killer(NK)cell exhaustion.Cytokine array and RNA sequencing analysis were combined to indicate the iron-binding protein LIPOCALIN-2(LCN-2)as a key factor in remodeling tumor microenvironments in the HCC-on-a-chip.LCN-2 targeted therapy demonstrated robust anti-tumor effects both in vitro 3D biomimetic chip and in vivo mouse model,including angiogenesis inhibition,sorafenib sensitivity promotion and NK-cell cytotoxicity enhancement.Taken together,the microfluidic platform exhibited obvious advantages in mimicking functional characteristics of tumor microenvironments and developing targeted therapies.
基金supported by the following grants:the National Natural Science Foundation of China(91749108,31671424,and 81322004 to H.M.,81200036 to M.L.,and 81102006 to J.Z.)the Science and Technology Research and Development Program of Shaanxi Province,China(2018SF-101 to N.M.and 2019SF-008 to M.L.)the Youth Innovation Team of Shaanxi Universities,China(to H.M.,Y.Y.,N.M.,Y.W.,and J.Z.)。
文摘Clinical cases have reported pulmonary arterial structural and functional abnormalities in patients with Kawasaki disease(KD);however,the underlying mechanisms are unclear.In this study,a KD rat model was established via the intraperitoneal injection of Lactobacillus casei cell wall extract(LCWE).The results showed that pulmonary arterial functional and structural abnormalities were observed in KD rats.Furthermore,proliferative endoplasmic reticulum stress(ER stress)was observed in the pulmonary arteries of KD rats.Notably,the level of lipocalin-2(Lcn 2),a trigger factor of inflammation,was remarkably elevated in the plasma and lung tissues of KD rats;increased Lcn 2 levels following LCWE stimulation may result from polymorphonuclear neutrophils(PMNs).Correspondingly,in cultured pulmonary artery smooth muscle cells(PASMCs),Lcn 2 markedly augmented the cleavage and nuclear localization of activating transcription factor-6(ATF6),upregulated the transcription of glucose regulated protein 78(GRP78)and neurite outgrowth inhibitor(NOGO),and promoted PASMCs proliferation.However,proapoptotic C/EBP homologous protein(CHOP)and caspase 12 levels were not elevated.Treatment with 4-phenyl butyric acid(4-PBA,a specific inhibitor of ER stress)inhibited PASMCs proliferation induced by Lcn 2 and attenuated pulmonary arterial abnormalities and right ventricular hypertrophy and reduced right ventricular systolic pressure in KD rats.In conclusion,Lcn 2 remarkably facilitates proliferative ER stress in PASMCs,which probably accounts for KD-related pulmonary arterial abnormalities.
