目的:探讨二甲双胍通过阻断Src酪氨酸激酶(Src)/信号转导及转录激蛋白3(STAT3)信号通路诱导胃癌MGC-803细胞凋亡的机制。方法:设MGC803细胞组、氟尿嘧啶组(100.0μg·ml^-1)和二甲双胍低(80.0μg·ml^-1)、高(160.0μg·ml^...目的:探讨二甲双胍通过阻断Src酪氨酸激酶(Src)/信号转导及转录激蛋白3(STAT3)信号通路诱导胃癌MGC-803细胞凋亡的机制。方法:设MGC803细胞组、氟尿嘧啶组(100.0μg·ml^-1)和二甲双胍低(80.0μg·ml^-1)、高(160.0μg·ml^-1)剂量组,各组细胞置于CO2培养箱(37℃、5%CO2、2%O2、93%N2)。以上各组每孔设6个平行样,培养72 h。培养结束后,细胞计数试剂盒-8(CCK-8)测定细胞活力,结晶紫染色测定单克隆形成数目,Transwell室测定侵袭能力,FACScan流式细胞仪测定细胞凋亡水平,实时荧光定量PCR(RT-PCR)及蛋白质印迹法(West Blot)测定p-SRC、p-STAT3 m RNA和蛋白水平。结果:氟尿嘧啶组和二甲双胍组的吸光度(A)值、存活率水平、克隆形成数目、穿膜数、p-SRC、p-STAT3 m RNA及蛋白表达水平显著低于MGC803细胞组(P<0.05),凋亡率显著高于MGC803细胞组(P<0.05)。二甲双胍低剂量组的A值、存活率水平、克隆形成数目、穿膜数、P-SRC、P-STAT3 m RNA、蛋白表达水平显著高于氟尿嘧啶组(P<0.05),凋亡率显著低于氟尿嘧啶组(P<0.05)。二甲双胍高剂量组与氟尿嘧啶组比较差异无统计学意义(P>0.05)。结论:二甲双胍能抑制胃癌MGC-803细胞增殖,促进胃癌MGC-803细胞凋亡;其机制与二甲双胍抑制P-SRC、P-STAT3 mRNA和蛋白的表达有关。展开更多
Background:Diabetic wounds pose significant clinical challenges due to impaired healing processes,often resulting in chronic,nonhealing ulcers.Asiaticoside(AC),a natural triterpene derivative from Centella asiatica,ha...Background:Diabetic wounds pose significant clinical challenges due to impaired healing processes,often resulting in chronic,nonhealing ulcers.Asiaticoside(AC),a natural triterpene derivative from Centella asiatica,has demonstrated notable anti-inflammatory and wound-healing properties.However,the synergistic effects of nitric oxide(NO)-a recognized promoter of wound healing-combined with AC in treating diabetic wounds remain inadequately explored.Methods:Ultraperformance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)was utilized to identify differential metabolites and dysregulated metabolic pathways associated with diabetic wounds.Molecular docking analyses were conducted to confirm the binding affinity of AC to key therapeutic targets.The effects of asiaticoside-nitric oxide hydrogel(ACNO)on gene and protein expression were evaluated using reverse transcription-quantitative polymerase chain reaction(RT-qPCR)and western blotting.In vitro experiments using sarcoma(SRC)agonists and inhibitors were performed to investigate the impact of ACNO therapy on the expression of SRC,STAT3,and other proteins in HaCaT cells.Results:Metabolomic profiling revealed that diabetic wounds in mice exhibited marked metabolic dysregulation,which was attenuated by ACNO treatment.Key metabolites modulated by ACNO included mandelic acid,lactic acid,and 3-hydroxyisovaleric acid.The primary metabolic pathways involved were methyl histidine metabolism and the malate-aspartate shuttle.Immunofluorescence staining confirmed that ACNO therapy enhanced angiogenesis,promoted cellular proliferation,and facilitated diabetic wound closure.RT-qPCR data demonstrated that ACNO regulated the transcription of critical genes(SRC,STAT3,EGFR,and VEGFA).Notably,ACNO attenuated SRC/STAT3 pathway activation while concurrently upregulating EGFR and VEGFA expression.Conclusions:These findings emphasize the therapeutic potential of ACNO hydrogel in diabetic wound healing through the modulation of metabolic pathways and the SRC/STAT3 signaling axis.By correlating altered metabolites with molecular targets,this study elucidates the pharmacodynamic foundation for ACNO's preclinical application and provides valuable insights into the development of targeted therapies for diabetic wound management.展开更多
文摘目的:探讨二甲双胍通过阻断Src酪氨酸激酶(Src)/信号转导及转录激蛋白3(STAT3)信号通路诱导胃癌MGC-803细胞凋亡的机制。方法:设MGC803细胞组、氟尿嘧啶组(100.0μg·ml^-1)和二甲双胍低(80.0μg·ml^-1)、高(160.0μg·ml^-1)剂量组,各组细胞置于CO2培养箱(37℃、5%CO2、2%O2、93%N2)。以上各组每孔设6个平行样,培养72 h。培养结束后,细胞计数试剂盒-8(CCK-8)测定细胞活力,结晶紫染色测定单克隆形成数目,Transwell室测定侵袭能力,FACScan流式细胞仪测定细胞凋亡水平,实时荧光定量PCR(RT-PCR)及蛋白质印迹法(West Blot)测定p-SRC、p-STAT3 m RNA和蛋白水平。结果:氟尿嘧啶组和二甲双胍组的吸光度(A)值、存活率水平、克隆形成数目、穿膜数、p-SRC、p-STAT3 m RNA及蛋白表达水平显著低于MGC803细胞组(P<0.05),凋亡率显著高于MGC803细胞组(P<0.05)。二甲双胍低剂量组的A值、存活率水平、克隆形成数目、穿膜数、P-SRC、P-STAT3 m RNA、蛋白表达水平显著高于氟尿嘧啶组(P<0.05),凋亡率显著低于氟尿嘧啶组(P<0.05)。二甲双胍高剂量组与氟尿嘧啶组比较差异无统计学意义(P>0.05)。结论:二甲双胍能抑制胃癌MGC-803细胞增殖,促进胃癌MGC-803细胞凋亡;其机制与二甲双胍抑制P-SRC、P-STAT3 mRNA和蛋白的表达有关。
基金supported by the National Natural Science Foundation of China(No.82160770,82460792)the Outstanding Young Scientific and Technological Talents Project of Guizhou Province(No.2021-5639)+1 种基金the Zunyi Science and Technology Talent Platform Carrier Construction Project(No.ZSKRPT2023-1)scholarships from the China Scholarship Council(No.CSC-202008520012).
文摘Background:Diabetic wounds pose significant clinical challenges due to impaired healing processes,often resulting in chronic,nonhealing ulcers.Asiaticoside(AC),a natural triterpene derivative from Centella asiatica,has demonstrated notable anti-inflammatory and wound-healing properties.However,the synergistic effects of nitric oxide(NO)-a recognized promoter of wound healing-combined with AC in treating diabetic wounds remain inadequately explored.Methods:Ultraperformance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)was utilized to identify differential metabolites and dysregulated metabolic pathways associated with diabetic wounds.Molecular docking analyses were conducted to confirm the binding affinity of AC to key therapeutic targets.The effects of asiaticoside-nitric oxide hydrogel(ACNO)on gene and protein expression were evaluated using reverse transcription-quantitative polymerase chain reaction(RT-qPCR)and western blotting.In vitro experiments using sarcoma(SRC)agonists and inhibitors were performed to investigate the impact of ACNO therapy on the expression of SRC,STAT3,and other proteins in HaCaT cells.Results:Metabolomic profiling revealed that diabetic wounds in mice exhibited marked metabolic dysregulation,which was attenuated by ACNO treatment.Key metabolites modulated by ACNO included mandelic acid,lactic acid,and 3-hydroxyisovaleric acid.The primary metabolic pathways involved were methyl histidine metabolism and the malate-aspartate shuttle.Immunofluorescence staining confirmed that ACNO therapy enhanced angiogenesis,promoted cellular proliferation,and facilitated diabetic wound closure.RT-qPCR data demonstrated that ACNO regulated the transcription of critical genes(SRC,STAT3,EGFR,and VEGFA).Notably,ACNO attenuated SRC/STAT3 pathway activation while concurrently upregulating EGFR and VEGFA expression.Conclusions:These findings emphasize the therapeutic potential of ACNO hydrogel in diabetic wound healing through the modulation of metabolic pathways and the SRC/STAT3 signaling axis.By correlating altered metabolites with molecular targets,this study elucidates the pharmacodynamic foundation for ACNO's preclinical application and provides valuable insights into the development of targeted therapies for diabetic wound management.
