Objective The goal of this study is to investigate the role and mechanism of endoplasmic reticulum stress and apoptosis regulated by thrombospondin 1(TSP1)in human renal tubular epithelial cells(HK-2 cells).Methods HK...Objective The goal of this study is to investigate the role and mechanism of endoplasmic reticulum stress and apoptosis regulated by thrombospondin 1(TSP1)in human renal tubular epithelial cells(HK-2 cells).Methods HK-2 cells were exposed to high concentrations of glucose(HG).The endoplasmic reticulum stress inhibitor 4-phenylbutyric acid(4-PBA)was administered by transfecting TSP1 or an empty vector to explore the mechanism of the endoplasmic reticulum response regulated by TSP1 and stress in renal cell apoptosis.The effects of TSP1 and 4-PBA on the proliferation and apoptosis of HK-2 cells under HG conditions were assessed using Cell counting kit-8 and flow cytometry.Western blotting was used to detect the apoptosis-and endoplasmic reticulum stress-related protein expression regulated by TSP1 and 4-PBA.Results HG treatment induced high cell apoptosis,abundantly expressed TSP1 level and restrained viability in HK-2 cells.Overexpression of TSP1 significantly inhibited the proliferation of and facilitated apoptosis of HK-2 cells under HG conditions.Administration of endoplasmic reticulum stress inhibitor 4-PBA after overexpression of TSP1 antagonized the inhibitory proliferation and promoted apoptosis rate in HG-triggered HK-2 cells induced by TSP1 overexpression.4-PBA treatment significantly hindered the expression of endoplasmic reticulum stress markers,such as PERK,ATF4,ATF6,p-eIF2α,IRE1,CHOP and XBP1,suggesting that the administration of 4-PBA was successful.Conclusion Overexpression of TSP1 activated endoplasmic reticulum stress by regulating the ATF6-CHOP axis.TSP1 restrained cell proliferation,and promoted apoptosis and endoplasmic reticulum stress by activating the ATF6-CHOP axis.展开更多
In this study, a combination of growth factors was used to induce bone marrow mesenchymal stem cells differentiation into neuron-like cells, in a broader attempt to observe the role of thrombospondin 1 in synapse form...In this study, a combination of growth factors was used to induce bone marrow mesenchymal stem cells differentiation into neuron-like cells, in a broader attempt to observe the role of thrombospondin 1 in synapse formation. Results showed that there was no significant difference in the differentiation rate of neuron-like cells between bone marrow mesenchymal stem cells with thrombospondin induction and those without. However, the cell shape was more complex and the neurites were dendritic, with unipolar, bipolar or multipolar morphologies, after induction with thrombospondin 1. The induced cells were similar in morphology to normal neurites. Immunohistochemical staining showed that the number of positive cells for postsynaptic density protein 95 and synaptophysin 1 protein was significantly increased after induction with thrombospondin 1. These findings indicate that thrombospondin 1 promotes synapse formation in neuron-like cells that are differentiated from bone marrow mesenchymal stem cells.展开更多
Background:Inhibitor of NF-κB kinase-interacting protein(IKIP)is known to promote proliferation of glioblastoma(GBM)cells,but how it affects migration and invasion by those cells is unclear.Methods:We compared levels...Background:Inhibitor of NF-κB kinase-interacting protein(IKIP)is known to promote proliferation of glioblastoma(GBM)cells,but how it affects migration and invasion by those cells is unclear.Methods:We compared levels of IKIP between glioma tissues and normal brain tissue in clinical samples and public databases.We examined the effects of IKIP overexpression and knockdown on the migration and invasion of GBM using transwell and wound healing assays,and we compared the transcriptomes under these different conditions to identify the molecular mechanisms involved.Results:Based on data from our clinical samples and from public databases,IKIP was overexpressed in GBM tumors,and its expression level correlated inversely with survival.IKIP overexpression in GBM cells inhibited migration and invasion in transwell and wound healing assays,whereas IKIP knockdown exerted the opposite effects.IKIP overexpression in GBM cells that were injected into mouse brain promoted tumor growth but inhibited tumor invasion of surrounding tissue.The effects of IKIP were associated with downregulation of THBS1 mRNA and concomitant inhibition of THBS1/FAK signaling.Conclusions:IKIP inhibits THBS1/FAK signaling to suppress migration and invasion of GBM cells.展开更多
目的探究H型原发性高血压(高血压)患者血清诱骗受体3(decoy receptor 3,DcR3)、含1型血小板反应蛋白基序的去整合素金属蛋白酶13(A disintegrin and metalloproteinase with A thrombospondin type 1 motif member 13,ADAMTS13)浓度与...目的探究H型原发性高血压(高血压)患者血清诱骗受体3(decoy receptor 3,DcR3)、含1型血小板反应蛋白基序的去整合素金属蛋白酶13(A disintegrin and metalloproteinase with A thrombospondin type 1 motif member 13,ADAMTS13)浓度与其心血管功能及预后的关系。方法选取大庆市人民医院2020年6月至2022年6月收治的132例高血压患者作为观察对象,根据同型半胱氨酸(homocysteine,Hcy)浓度分为非H型高血压组40例和H型高血压组92例,根据预后情况将H型高血压患者分为预后良好组和预后不良组,并选择同期来大庆市人民医院健康体检的成年人70名作为对照组。采用酶联免疫吸附试验法检测受试者血清中DcR3、ADAMTS13浓度,Pearson法分析血清中DcR3、ADAMTS13浓度与心血管功能指标的相关性,多因素Logistic回归分析H型高血压患者1年预后不良的影响因素。绘制受试者工作特征曲线(receiver operating characteristic curve,ROC)分析血清DcR3、ADAMTS13浓度对H型高血压患者1年预后不良的预测价值。结果与对照组[(122.28±32.34)mmHg(1 mmHg=0.133 kPa)、(48.16±8.65)mmHg、(8.59±1.25)mm、(118.34±34.25)g/m2、(1.48±0.34)g/L、(57.15±14.94)mg/L、(1.45±0.31)、70.28%±15.21%]比较,H型高血压组患者的收缩压[(139.35±38.21)mmHg]、脉压[(57.37±11.75)mmHg]、左心室后壁厚度(posterior wall thickness,PWT)[(11.69±2.00)mm]以及左心室质量指数(left ventricular mass index,LVMI)[(148.54±38.22)g/m2]显著升高,DcR3[(0.74±0.19)g/L]、ADAMTS13浓度[(14.13±4.62)mg/L]、二尖瓣舒张早期血流峰值/二尖瓣舒张晚期血流峰值(E-peak to A-peak of the mitral flow spectrum,E/A)(0.65±0.13)、左心室射血分数(left ventricular ejection fraction,LVEF)(64.26%±12.75%)显著降低,差异有统计学意义(P<0.05);与非H型高血压组患者组比较,H型高血压组患者的DcR3、ADAMTS13浓度及E/A显著降低,LVMI显著升高,差异有统计学意义(P<0.05)。H型高血压组患者血清中DcR3、ADAMTS13浓度均与收缩压、脉压和LVMI呈负相关(P<0.05),而与E/A、LVEF呈正相关(P<0.05)。预后不良组患者的年龄显著高于预后良好组,E/A(0.38±0.07)、DcR3[(0.45±0.13)g/L]、ADAMTS13浓度[(8.45±2.11)mg/L]显著低于预后良好组[0.75±0.11、(0.85±0.27)g/L、(16.25±4.85)mg/L],差异有统计学意义(P<0.05)。DcR3、ADAMTS13是H型高血压患者预后不良的保护因素(P<0.05)。血清DcR3、ADAMTS13浓度单独及二者联合预测H型高血压患者1年发生预后不良的曲线下面积(area under the curve,AUC)分别为0.906、0.844、0.950。结论H型高血压疾病患者血清DcR3、ADAMTS13浓度降低,与心血管功能及预后密切相关,对该疾病的预后评估有重要价值。展开更多
目的分析超声心动图联合血清凝血酶敏感蛋白1(thrombospondin-1,TSP-1)、甲壳质酶蛋白40(chitinase protein 40,YKL-40)浓度检测对高血压性心脏病患者的诊断价值。方法选择2021年1月至2022年6月四川友谊医院收治的高血压性心脏病患者98...目的分析超声心动图联合血清凝血酶敏感蛋白1(thrombospondin-1,TSP-1)、甲壳质酶蛋白40(chitinase protein 40,YKL-40)浓度检测对高血压性心脏病患者的诊断价值。方法选择2021年1月至2022年6月四川友谊医院收治的高血压性心脏病患者98例(心脏病组)进行研究,患者入组均接受超声心动图检查。酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)检测血清TSP-1、YKL-40浓度。Pearson法分析高血压性心脏病患者血清TSP-1、YKL-40浓度与超声心动图各项参数的相关性,采用受试者工作特征曲线(receiver operating characteristic curve,ROC)评估血清TSP-1、YKL-40浓度诊断高血压性心脏病的可靠性。结果与对照组相比,心脏病组患者的收缩压、舒张压、左心房前后直径(left atrial anteroposterior diameter,LAD)、左心室收缩末期内径(left ventricular end systolic diameter,LVESD)、左心房内径指数(left atrial inner diameter index,LADI)、室间隔舒张末期厚度(end-diastolic interventricular septum thickness,IVST)、左心室舒张末期内径(left ventricular end-diastolic diameter,LVEDD)、左心室后壁舒张末期厚度(left ventricular posterior wall end-diastolic thickness,LVPWT)、左心室舒张晚期峰值流速(diastolic advanced stage filling peak rate of flow,VA)值较高,二尖瓣血流舒张早期左心室充盈峰速(peak velocity of left ventricular filling during early diastole,VE)值、峰值速度比率(ratio of E velocity to A velocity,E/A)较低,差异有统计学意义(P<0.05)。与对照组相比,心脏病组患者血清TSP-1、YKL-40浓度较高,差异有统计学意义(P<0.05)。Pearson法分析显示,血清TSP-1、YKL-40浓度与LAD、LVESD、LADI、IVST、LVEDD、LVPWT、VA值均呈正相关(P<0.05),与VE值、E/A均呈负相关(P<0.05)。超声心动图评估患者发生高血压性心脏病的准确度为88.00%,血清TSP-1、YKL-40浓度对高血压性心脏病诊断的准确度为81.00%、82.50%。超声心动图联合血清TSP-1、YKL-40浓度对高血压性心脏病诊断的准确度为91.50%,其灵敏度为95.92%,特异度为87.25%。结论高血压性心脏病患者血清中TSP-1、YKL-40浓度升高,血清TSP-1、YKL-40浓度对高血压性心脏病有一定诊断价值,超声心动图联合血清TSP-1、YKL-40浓度可明显提高高血压性心脏病诊断的准确性。展开更多
Purpose: To investigate the expression of thrombospondin 1 (TSP-1) in retinal pigment epithelium (RPE) and choroidal neovascular membranes (CNVMs) from patients with age-related macular degeneration (AMD). Methods: Ti...Purpose: To investigate the expression of thrombospondin 1 (TSP-1) in retinal pigment epithelium (RPE) and choroidal neovascular membranes (CNVMs) from patients with age-related macular degeneration (AMD). Methods: Tissue sections from normal human fetal and adult eyes and surgically removed CNVMs were immunostained for TSP-1 localization. Polymerase chain reaction and Western blotting were used to analyze TSP-1 mRNA and protein from human RPE cells, respectively. TSP-1 in the supernatant of cultured RPE cells and eye explants were measured using enzyme-linked immunosorbent assay. MTT assay was used to evaluate the RPE survival after TSP-1 treatment. Results: The strongest immunostaining for TSP-1 was observed in the RPE monolayer around drusen in early AMD. The intensity of TSP-1 staining in normal eye sections was much weaker than that of early AMD and CNVM. TSP-1 mRNA was positive in cultured fetal and adult RPE cells. There was increasing secretion of TSP-1 into the supernatant of cultured RPE and eye explants. The specific band of TSP-1 was identified by Western blot. No significant inhibition of RPE survival was found with the exposure to TSP-1. Conclusions: TSP-1 expression in drusen and CNVM was upregulated and associated with RPE monolayer. TSP-1 may be a natural negative regulator for choroidal neovascularization.展开更多
基金This study was supported by a grant from Chinese Society of Nephrology(No.14050430580).
文摘Objective The goal of this study is to investigate the role and mechanism of endoplasmic reticulum stress and apoptosis regulated by thrombospondin 1(TSP1)in human renal tubular epithelial cells(HK-2 cells).Methods HK-2 cells were exposed to high concentrations of glucose(HG).The endoplasmic reticulum stress inhibitor 4-phenylbutyric acid(4-PBA)was administered by transfecting TSP1 or an empty vector to explore the mechanism of the endoplasmic reticulum response regulated by TSP1 and stress in renal cell apoptosis.The effects of TSP1 and 4-PBA on the proliferation and apoptosis of HK-2 cells under HG conditions were assessed using Cell counting kit-8 and flow cytometry.Western blotting was used to detect the apoptosis-and endoplasmic reticulum stress-related protein expression regulated by TSP1 and 4-PBA.Results HG treatment induced high cell apoptosis,abundantly expressed TSP1 level and restrained viability in HK-2 cells.Overexpression of TSP1 significantly inhibited the proliferation of and facilitated apoptosis of HK-2 cells under HG conditions.Administration of endoplasmic reticulum stress inhibitor 4-PBA after overexpression of TSP1 antagonized the inhibitory proliferation and promoted apoptosis rate in HG-triggered HK-2 cells induced by TSP1 overexpression.4-PBA treatment significantly hindered the expression of endoplasmic reticulum stress markers,such as PERK,ATF4,ATF6,p-eIF2α,IRE1,CHOP and XBP1,suggesting that the administration of 4-PBA was successful.Conclusion Overexpression of TSP1 activated endoplasmic reticulum stress by regulating the ATF6-CHOP axis.TSP1 restrained cell proliferation,and promoted apoptosis and endoplasmic reticulum stress by activating the ATF6-CHOP axis.
基金supported by the Natural Science Foundation of Guangdong Province, No. S2011010004096the Medical Scientific Research Foundation of Guangdong Province, No. A2010431 A2009477
文摘In this study, a combination of growth factors was used to induce bone marrow mesenchymal stem cells differentiation into neuron-like cells, in a broader attempt to observe the role of thrombospondin 1 in synapse formation. Results showed that there was no significant difference in the differentiation rate of neuron-like cells between bone marrow mesenchymal stem cells with thrombospondin induction and those without. However, the cell shape was more complex and the neurites were dendritic, with unipolar, bipolar or multipolar morphologies, after induction with thrombospondin 1. The induced cells were similar in morphology to normal neurites. Immunohistochemical staining showed that the number of positive cells for postsynaptic density protein 95 and synaptophysin 1 protein was significantly increased after induction with thrombospondin 1. These findings indicate that thrombospondin 1 promotes synapse formation in neuron-like cells that are differentiated from bone marrow mesenchymal stem cells.
基金supported by the National Natural Science Foundation of China(82002638)the National Natural Science Foundation of Sichuan Province(2023NSFSC0734).
文摘Background:Inhibitor of NF-κB kinase-interacting protein(IKIP)is known to promote proliferation of glioblastoma(GBM)cells,but how it affects migration and invasion by those cells is unclear.Methods:We compared levels of IKIP between glioma tissues and normal brain tissue in clinical samples and public databases.We examined the effects of IKIP overexpression and knockdown on the migration and invasion of GBM using transwell and wound healing assays,and we compared the transcriptomes under these different conditions to identify the molecular mechanisms involved.Results:Based on data from our clinical samples and from public databases,IKIP was overexpressed in GBM tumors,and its expression level correlated inversely with survival.IKIP overexpression in GBM cells inhibited migration and invasion in transwell and wound healing assays,whereas IKIP knockdown exerted the opposite effects.IKIP overexpression in GBM cells that were injected into mouse brain promoted tumor growth but inhibited tumor invasion of surrounding tissue.The effects of IKIP were associated with downregulation of THBS1 mRNA and concomitant inhibition of THBS1/FAK signaling.Conclusions:IKIP inhibits THBS1/FAK signaling to suppress migration and invasion of GBM cells.
文摘目的探究H型原发性高血压(高血压)患者血清诱骗受体3(decoy receptor 3,DcR3)、含1型血小板反应蛋白基序的去整合素金属蛋白酶13(A disintegrin and metalloproteinase with A thrombospondin type 1 motif member 13,ADAMTS13)浓度与其心血管功能及预后的关系。方法选取大庆市人民医院2020年6月至2022年6月收治的132例高血压患者作为观察对象,根据同型半胱氨酸(homocysteine,Hcy)浓度分为非H型高血压组40例和H型高血压组92例,根据预后情况将H型高血压患者分为预后良好组和预后不良组,并选择同期来大庆市人民医院健康体检的成年人70名作为对照组。采用酶联免疫吸附试验法检测受试者血清中DcR3、ADAMTS13浓度,Pearson法分析血清中DcR3、ADAMTS13浓度与心血管功能指标的相关性,多因素Logistic回归分析H型高血压患者1年预后不良的影响因素。绘制受试者工作特征曲线(receiver operating characteristic curve,ROC)分析血清DcR3、ADAMTS13浓度对H型高血压患者1年预后不良的预测价值。结果与对照组[(122.28±32.34)mmHg(1 mmHg=0.133 kPa)、(48.16±8.65)mmHg、(8.59±1.25)mm、(118.34±34.25)g/m2、(1.48±0.34)g/L、(57.15±14.94)mg/L、(1.45±0.31)、70.28%±15.21%]比较,H型高血压组患者的收缩压[(139.35±38.21)mmHg]、脉压[(57.37±11.75)mmHg]、左心室后壁厚度(posterior wall thickness,PWT)[(11.69±2.00)mm]以及左心室质量指数(left ventricular mass index,LVMI)[(148.54±38.22)g/m2]显著升高,DcR3[(0.74±0.19)g/L]、ADAMTS13浓度[(14.13±4.62)mg/L]、二尖瓣舒张早期血流峰值/二尖瓣舒张晚期血流峰值(E-peak to A-peak of the mitral flow spectrum,E/A)(0.65±0.13)、左心室射血分数(left ventricular ejection fraction,LVEF)(64.26%±12.75%)显著降低,差异有统计学意义(P<0.05);与非H型高血压组患者组比较,H型高血压组患者的DcR3、ADAMTS13浓度及E/A显著降低,LVMI显著升高,差异有统计学意义(P<0.05)。H型高血压组患者血清中DcR3、ADAMTS13浓度均与收缩压、脉压和LVMI呈负相关(P<0.05),而与E/A、LVEF呈正相关(P<0.05)。预后不良组患者的年龄显著高于预后良好组,E/A(0.38±0.07)、DcR3[(0.45±0.13)g/L]、ADAMTS13浓度[(8.45±2.11)mg/L]显著低于预后良好组[0.75±0.11、(0.85±0.27)g/L、(16.25±4.85)mg/L],差异有统计学意义(P<0.05)。DcR3、ADAMTS13是H型高血压患者预后不良的保护因素(P<0.05)。血清DcR3、ADAMTS13浓度单独及二者联合预测H型高血压患者1年发生预后不良的曲线下面积(area under the curve,AUC)分别为0.906、0.844、0.950。结论H型高血压疾病患者血清DcR3、ADAMTS13浓度降低,与心血管功能及预后密切相关,对该疾病的预后评估有重要价值。
文摘目的分析超声心动图联合血清凝血酶敏感蛋白1(thrombospondin-1,TSP-1)、甲壳质酶蛋白40(chitinase protein 40,YKL-40)浓度检测对高血压性心脏病患者的诊断价值。方法选择2021年1月至2022年6月四川友谊医院收治的高血压性心脏病患者98例(心脏病组)进行研究,患者入组均接受超声心动图检查。酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)检测血清TSP-1、YKL-40浓度。Pearson法分析高血压性心脏病患者血清TSP-1、YKL-40浓度与超声心动图各项参数的相关性,采用受试者工作特征曲线(receiver operating characteristic curve,ROC)评估血清TSP-1、YKL-40浓度诊断高血压性心脏病的可靠性。结果与对照组相比,心脏病组患者的收缩压、舒张压、左心房前后直径(left atrial anteroposterior diameter,LAD)、左心室收缩末期内径(left ventricular end systolic diameter,LVESD)、左心房内径指数(left atrial inner diameter index,LADI)、室间隔舒张末期厚度(end-diastolic interventricular septum thickness,IVST)、左心室舒张末期内径(left ventricular end-diastolic diameter,LVEDD)、左心室后壁舒张末期厚度(left ventricular posterior wall end-diastolic thickness,LVPWT)、左心室舒张晚期峰值流速(diastolic advanced stage filling peak rate of flow,VA)值较高,二尖瓣血流舒张早期左心室充盈峰速(peak velocity of left ventricular filling during early diastole,VE)值、峰值速度比率(ratio of E velocity to A velocity,E/A)较低,差异有统计学意义(P<0.05)。与对照组相比,心脏病组患者血清TSP-1、YKL-40浓度较高,差异有统计学意义(P<0.05)。Pearson法分析显示,血清TSP-1、YKL-40浓度与LAD、LVESD、LADI、IVST、LVEDD、LVPWT、VA值均呈正相关(P<0.05),与VE值、E/A均呈负相关(P<0.05)。超声心动图评估患者发生高血压性心脏病的准确度为88.00%,血清TSP-1、YKL-40浓度对高血压性心脏病诊断的准确度为81.00%、82.50%。超声心动图联合血清TSP-1、YKL-40浓度对高血压性心脏病诊断的准确度为91.50%,其灵敏度为95.92%,特异度为87.25%。结论高血压性心脏病患者血清中TSP-1、YKL-40浓度升高,血清TSP-1、YKL-40浓度对高血压性心脏病有一定诊断价值,超声心动图联合血清TSP-1、YKL-40浓度可明显提高高血压性心脏病诊断的准确性。
文摘Purpose: To investigate the expression of thrombospondin 1 (TSP-1) in retinal pigment epithelium (RPE) and choroidal neovascular membranes (CNVMs) from patients with age-related macular degeneration (AMD). Methods: Tissue sections from normal human fetal and adult eyes and surgically removed CNVMs were immunostained for TSP-1 localization. Polymerase chain reaction and Western blotting were used to analyze TSP-1 mRNA and protein from human RPE cells, respectively. TSP-1 in the supernatant of cultured RPE cells and eye explants were measured using enzyme-linked immunosorbent assay. MTT assay was used to evaluate the RPE survival after TSP-1 treatment. Results: The strongest immunostaining for TSP-1 was observed in the RPE monolayer around drusen in early AMD. The intensity of TSP-1 staining in normal eye sections was much weaker than that of early AMD and CNVM. TSP-1 mRNA was positive in cultured fetal and adult RPE cells. There was increasing secretion of TSP-1 into the supernatant of cultured RPE and eye explants. The specific band of TSP-1 was identified by Western blot. No significant inhibition of RPE survival was found with the exposure to TSP-1. Conclusions: TSP-1 expression in drusen and CNVM was upregulated and associated with RPE monolayer. TSP-1 may be a natural negative regulator for choroidal neovascularization.
