OBJECTIVE:To investigate the impact of Shenhua tablet(肾华片,SHT)on renal macrophage polarization and renal injury in mice with diabetic kidney disease(DKD)and to explore the potential mechanism involving the hypoxia-...OBJECTIVE:To investigate the impact of Shenhua tablet(肾华片,SHT)on renal macrophage polarization and renal injury in mice with diabetic kidney disease(DKD)and to explore the potential mechanism involving the hypoxia-inducible factor-1α(HIF-1α)and pyruvate kinase M2(PKM2)signaling pathway,along with the glycolysis metabolism pathway.METHODS:The animals were divided into the following groups:Model,Control,dapagliflozin,SHT low-dose,SHT medium-dose,and SHT high-dose.We assessed 24-hour urine protein(24 h-UTP)levels,urinary albuminto-creatinine ratio,and regularly monitored fasting blood glucose during the treatment period.After treatment,we examined renal tissue structure,renal function(urea nitrogen,uric acid,creatinine,cystatin C,β2-microglobulin),and glycolysis in renal macrophages.Additionally,we observed macrophage polarization in renal tissue and measured inflammatory factors(tumor necrosis factor-α,interleukin-1β,interleukin-6,interleukin-10,monocyte chemoattractant protein-1)to assess the immunoinflammatory status of the renal tissue.Finally,we investigated the expression of the HIF-1α/PKM2 signaling pathway in macrophages to explore its role in the glycolysis process.RESULTS:SHT shows a beneficial effect in treating DKD by reducing 24 h-UTP,regulating blood glucose levels,improving renal tissue structure,protecting renal function,inhibiting macrophage glycolysis,reducing macrophage transformation to the M1 state,and suppressing the expression of the HIF-1α/PKM2 signaling pathway.CONCLUSION:SHT may exert renoprotective effects by inhibiting macrophage glycolysis via the HIF-1α/PKM2 signaling pathway.This inhibition decreases macrophage M1 polarization and reduces immunoinflammatory injury in the renal tissue of DKD mice.展开更多
目的:研究PKM2在非小细胞肺癌组织中的表达及其与放疗敏感性的关系。方法:选取2011年5月至2013年10月四川省人民医院非小细胞肺癌120例患者,术后进行2个月的放射治疗,按照疗效将患者分为4组。RT-PCR检测患者放疗前手术切除的肿瘤组织和...目的:研究PKM2在非小细胞肺癌组织中的表达及其与放疗敏感性的关系。方法:选取2011年5月至2013年10月四川省人民医院非小细胞肺癌120例患者,术后进行2个月的放射治疗,按照疗效将患者分为4组。RT-PCR检测患者放疗前手术切除的肿瘤组织和癌旁组织中PKM2 m RNA表达水平。Western blot和免疫组织化学检测肿瘤组织和癌旁组织中PKM2蛋白表达。RT-PCR和Western blot方法检测不同疗效组PKM2表达水平。结果:非小细胞肺癌放疗2个月后,治疗有效率为49.2%。PKM2在非小细胞肺癌癌组织中表达显著高于相应的癌旁组织。PKM2表达水平与患者放疗疗效呈负相关。结论:PKM2在非小细胞肺癌中表达高于癌旁组织,PKM2低表达患者对放疗敏感性高。展开更多
目的:探究TRIM2通过泛素化修饰PKM2调控肝癌细胞糖酵解的机制。方法:TCGA和CPTAC数据库分析TRIM2基因及蛋白表达水平,并结合病人生存状态进行相关性分析。构建HepG2-shNC和HepG2-shTRIM2细胞株,细胞能量代谢仪检测两组细胞胞外酸化率和...目的:探究TRIM2通过泛素化修饰PKM2调控肝癌细胞糖酵解的机制。方法:TCGA和CPTAC数据库分析TRIM2基因及蛋白表达水平,并结合病人生存状态进行相关性分析。构建HepG2-shNC和HepG2-shTRIM2细胞株,细胞能量代谢仪检测两组细胞胞外酸化率和耗氧率,并测定细胞葡萄糖吸收率、乳酸产生量、ATP水平。免疫共沉淀验证TRIM2和PKM2在HepG2、HEK293T细胞中相互作用,免疫荧光检测PKM2和TRIM2共定位。免疫共沉淀检测在HepG2/HEK293T细胞中敲低/过表达TRIM2后PKM2的泛素化修饰水平,体外酶催化反应检测在HepG2/HEK293T细胞中敲低/过表达TRIM2后PKM2酶活性。Transwell小室法检测HepG2-shNC和HepG2-shTRIM2细胞侵袭和迁移能力。皮下移植瘤实验检测HepG2-shNC和HepG2-shTRIM2细胞体内成瘤能力。结果:TCGA和CPTAC数据库分析发现TRIM2在肝癌组织中mRNA及蛋白表达水平均升高,高TRIM2表达预示差的患者生存状态。TRIM2表达水平与糖酵解通路相关性高,敲低TRIM2可抑制HepG2胞外酸化率,提高耗氧率,细胞葡萄糖吸收率[(100.0000±2.0000)%vs(46.6667±8.1445)%]、乳酸产生量[(99.0000±3.6055)%vs(43.6667±10.01665)%]、ATP水平[(97.0000±3.6055)%vs(57.3333±6.6583)%]均显著下调。TRIM2与PKM2在HepG2细胞中相互作用且蛋白共定位。TRIM2抑制后HepG2细胞中PKM2泛素化水平升高,PKM2酶活性降低;在HEK293T细胞中表达Myc-TRIM2,下调PKM2蛋白泛素化修饰水平,提高PKM2酶活性。敲低TRIM2抑制HepG2细胞侵袭能力(1 vs 0.5067±0.1168)、迁移能力(1 vs 0.5133±0.08622),抑制细胞体内成瘤体积[(529.3333±29.28026)mm 3 vs(281.6667±25.6969)mm 3]和重量[(360.3333±42.7161)mg vs(172.1667±50.2849)mg]。结论:TRIM2在肝癌组织中高表达,通过去泛素化修饰PKM2促进其酶活性,进而调控肿瘤细胞糖酵解能力。展开更多
基金National Natural Science Foundation of China:Basic Research on the Mechanism of Organ Immune Damage and the Diagnosis and Treatment of Integrated Traditional Chinese and Western Medicine(No.32141005)。
文摘OBJECTIVE:To investigate the impact of Shenhua tablet(肾华片,SHT)on renal macrophage polarization and renal injury in mice with diabetic kidney disease(DKD)and to explore the potential mechanism involving the hypoxia-inducible factor-1α(HIF-1α)and pyruvate kinase M2(PKM2)signaling pathway,along with the glycolysis metabolism pathway.METHODS:The animals were divided into the following groups:Model,Control,dapagliflozin,SHT low-dose,SHT medium-dose,and SHT high-dose.We assessed 24-hour urine protein(24 h-UTP)levels,urinary albuminto-creatinine ratio,and regularly monitored fasting blood glucose during the treatment period.After treatment,we examined renal tissue structure,renal function(urea nitrogen,uric acid,creatinine,cystatin C,β2-microglobulin),and glycolysis in renal macrophages.Additionally,we observed macrophage polarization in renal tissue and measured inflammatory factors(tumor necrosis factor-α,interleukin-1β,interleukin-6,interleukin-10,monocyte chemoattractant protein-1)to assess the immunoinflammatory status of the renal tissue.Finally,we investigated the expression of the HIF-1α/PKM2 signaling pathway in macrophages to explore its role in the glycolysis process.RESULTS:SHT shows a beneficial effect in treating DKD by reducing 24 h-UTP,regulating blood glucose levels,improving renal tissue structure,protecting renal function,inhibiting macrophage glycolysis,reducing macrophage transformation to the M1 state,and suppressing the expression of the HIF-1α/PKM2 signaling pathway.CONCLUSION:SHT may exert renoprotective effects by inhibiting macrophage glycolysis via the HIF-1α/PKM2 signaling pathway.This inhibition decreases macrophage M1 polarization and reduces immunoinflammatory injury in the renal tissue of DKD mice.
文摘目的:研究PKM2在非小细胞肺癌组织中的表达及其与放疗敏感性的关系。方法:选取2011年5月至2013年10月四川省人民医院非小细胞肺癌120例患者,术后进行2个月的放射治疗,按照疗效将患者分为4组。RT-PCR检测患者放疗前手术切除的肿瘤组织和癌旁组织中PKM2 m RNA表达水平。Western blot和免疫组织化学检测肿瘤组织和癌旁组织中PKM2蛋白表达。RT-PCR和Western blot方法检测不同疗效组PKM2表达水平。结果:非小细胞肺癌放疗2个月后,治疗有效率为49.2%。PKM2在非小细胞肺癌癌组织中表达显著高于相应的癌旁组织。PKM2表达水平与患者放疗疗效呈负相关。结论:PKM2在非小细胞肺癌中表达高于癌旁组织,PKM2低表达患者对放疗敏感性高。
文摘目的:探究TRIM2通过泛素化修饰PKM2调控肝癌细胞糖酵解的机制。方法:TCGA和CPTAC数据库分析TRIM2基因及蛋白表达水平,并结合病人生存状态进行相关性分析。构建HepG2-shNC和HepG2-shTRIM2细胞株,细胞能量代谢仪检测两组细胞胞外酸化率和耗氧率,并测定细胞葡萄糖吸收率、乳酸产生量、ATP水平。免疫共沉淀验证TRIM2和PKM2在HepG2、HEK293T细胞中相互作用,免疫荧光检测PKM2和TRIM2共定位。免疫共沉淀检测在HepG2/HEK293T细胞中敲低/过表达TRIM2后PKM2的泛素化修饰水平,体外酶催化反应检测在HepG2/HEK293T细胞中敲低/过表达TRIM2后PKM2酶活性。Transwell小室法检测HepG2-shNC和HepG2-shTRIM2细胞侵袭和迁移能力。皮下移植瘤实验检测HepG2-shNC和HepG2-shTRIM2细胞体内成瘤能力。结果:TCGA和CPTAC数据库分析发现TRIM2在肝癌组织中mRNA及蛋白表达水平均升高,高TRIM2表达预示差的患者生存状态。TRIM2表达水平与糖酵解通路相关性高,敲低TRIM2可抑制HepG2胞外酸化率,提高耗氧率,细胞葡萄糖吸收率[(100.0000±2.0000)%vs(46.6667±8.1445)%]、乳酸产生量[(99.0000±3.6055)%vs(43.6667±10.01665)%]、ATP水平[(97.0000±3.6055)%vs(57.3333±6.6583)%]均显著下调。TRIM2与PKM2在HepG2细胞中相互作用且蛋白共定位。TRIM2抑制后HepG2细胞中PKM2泛素化水平升高,PKM2酶活性降低;在HEK293T细胞中表达Myc-TRIM2,下调PKM2蛋白泛素化修饰水平,提高PKM2酶活性。敲低TRIM2抑制HepG2细胞侵袭能力(1 vs 0.5067±0.1168)、迁移能力(1 vs 0.5133±0.08622),抑制细胞体内成瘤体积[(529.3333±29.28026)mm 3 vs(281.6667±25.6969)mm 3]和重量[(360.3333±42.7161)mg vs(172.1667±50.2849)mg]。结论:TRIM2在肝癌组织中高表达,通过去泛素化修饰PKM2促进其酶活性,进而调控肿瘤细胞糖酵解能力。
