AIM:To explore the effect of silent information regulator factor 2-related enzyme 1(SIRT1)on modulating apoptosis of human lens epithelial cells(HLECs)and alleviating lens opacification of rats through suppressing end...AIM:To explore the effect of silent information regulator factor 2-related enzyme 1(SIRT1)on modulating apoptosis of human lens epithelial cells(HLECs)and alleviating lens opacification of rats through suppressing endoplasmic reticulum(ER)stress.METHODS:HLECs(SRA01/04)were treated with varying concentrations of tunicamycin(TM)for 24h,and the expression of SIRT1 and C/EBP homologous protein(CHOP)was assessed using real-time quantitative polymerase chain reaction(RT-PCR),Western blotting,and immunofluorescence.Cell morphology and proliferation was evaluated using an inverted microscope and cell counting kit-8(CCK-8)assay,respectively.In the SRA01/04 cell apoptosis model,which underwent siRNA transfection for SIRT1 knockdown and SRT1720 treatment for its activation,the expression levels of SIRT1,CHOP,glucose regulated protein 78(GRP78),and activating transcription factor 4(ATF4)were examined.The potential reversal of SIRT1 knockdown effects by 4-phenyl butyric acid(4-PBA;an ER stress inhibitor)was investigated.In vivo,age-related cataract(ARC)rat models were induced by sodium selenite injection,and the protective role of SIRT1,activated by SRT1720 intraperitoneal injections,was evaluated through morphology observation,hematoxylin and eosin(H&E)staining,Western blotting,and RT-PCR.RESULTS:SIRT1 expression was downregulated in TMinduced SRA01/04 cells.Besides,in SRA01/04 cells,both cell apoptosis and CHOP expression increased with the rising doses of TM.ER stress was stimulated by TM,as evidenced by the increased GRP78 and ATF4 in the SRA01/04 cell apoptosis model.Inhibition of SIRT1 by siRNA knockdown increased ER stress activation,whereas SRT1720 treatment had opposite results.4-PBA partly reverse the adverse effect of SIRT1 knockdown on apoptosis.In vivo,SRT1720 attenuated the lens opacification and weakened the ER stress activation in ARC rat models.CONCLUSION:SIRT1 plays a protective role against TM-induced apoptosis in HLECs and slows the progression of cataract in rats by inhibiting ER stress.These findings suggest a novel strategy for cataract treatment focused on targeting ER stress,highlighting the therapeutic potential of SIRT1 modulation in ARC development.展开更多
AIM:To examine the regulatory role of microRNA-204(miR-204)on silent information regulator 1(SIRT1)and vascular endothelial growth factor(VEGF)under highglucose-induced metabolic memory in human retinal pigment epithe...AIM:To examine the regulatory role of microRNA-204(miR-204)on silent information regulator 1(SIRT1)and vascular endothelial growth factor(VEGF)under highglucose-induced metabolic memory in human retinal pigment epithelial(hRPE)cells.METHODS:Cells were cultured with either normal(5 mmol/L)or high D-glucose(25 mmol/L)concentrations for 8d to establish control and high-glucose groups,respectively.To induce metabolic memory,cells were cultured with 25 mmol/L D-glucose for 4d followed by culture with 5 mmol/L D-glucose for 4d.In addition,exposed in 25 mmol/L D-glucose for 4d and then transfected with 100 nmol/L miR-204 control,miR-204 inhibitor or miR-204 mimic in 5 mmol/L D-glucose for 4d.Quantitative reverse transcription-polymerase chain reaction(RT-qPCR)was used to detect miR-204 mRNA levels.SIRT1 and VEGF protein levels were assessed by immunohistochemical and Western blot.Flow cytometry was used to investigate apoptosis rate.RESULTS:It was found that high glucose promoted miR-204 and VEGF expression,and inhibited SIRT1 activity,even after the return to normal glucose culture conditions.Upregulation of miR-204 promoted apoptosis inhibiting SIRT1 and increasing VEGF expression.However,downregulation of miR-204 produced the opposite effects.CONCLUSION:The study identifies that miR-204 is the upstream target of SIRT1and VEGF,and that miR-204 can protect hRPE cells from the damage caused by metabolic memory through increasing SIRT1 and inhibiting VEGF expression.展开更多
目的探讨miR-204和沉默信息调节因子1(silence information regulator 1,SIRT1)在结直肠癌组织中的表达情况及其临床价值。方法收集2018年5月至2020年6月嘉兴大学附属医院胃肠外科诊治的60例结直肠癌患者的癌组织标本和癌旁组织标本为...目的探讨miR-204和沉默信息调节因子1(silence information regulator 1,SIRT1)在结直肠癌组织中的表达情况及其临床价值。方法收集2018年5月至2020年6月嘉兴大学附属医院胃肠外科诊治的60例结直肠癌患者的癌组织标本和癌旁组织标本为研究对象,采用实时荧光定量聚合酶链反应检测miR-204及SIRT1的基因表达情况,Pearson分析比较miR-204与SIRT1基因的表达相关性。采用免疫组织化学SABC法检测SIRT1蛋白表达,并比较不同SIRT1蛋白表达与临床病理特征的关系。Kaplan-Meier法分析不同SIRT1蛋白表达的结直肠癌患者的生存差异。结果癌组织中miR-204基因mRNA表达显著低于癌旁组织(P<0.05),SIRT1基因mRNA表达显著高于癌旁组织(P<0.05)。癌组织中SIRT1蛋白表达阳性率显著高于癌旁组织(P<0.05)。Pearson相关分析显示miR-204与SIRT1基因mRNA在癌旁组织和癌组织中的表达均呈负相关(r=–0.647、–0.737,P<0.05)。SIRT1蛋白的表达与结直肠癌的分化水平、浸润层次、淋巴结转移与否及TNM分期相关(P<0.05),与患者的年龄、性别、肿瘤大小及肿瘤部位无关(P>0.05)。Kaplan-Meier分析显示癌组织中SIRT1阳性表达患者的生存率显著低于SIRT1阴性表达患者(χ^(2)=5.001,P=0.025)。结论结直肠癌组织中miR-204表达下调,SIRT1表达上调,二者可能通过相互影响共同促进结直肠癌的转移、侵袭,并影响患者预后。展开更多
基金Supported by National Natural Science Foundation for Young Scientists of China(No.82101097)National Natural Science Foundation of China(No.82070937).
文摘AIM:To explore the effect of silent information regulator factor 2-related enzyme 1(SIRT1)on modulating apoptosis of human lens epithelial cells(HLECs)and alleviating lens opacification of rats through suppressing endoplasmic reticulum(ER)stress.METHODS:HLECs(SRA01/04)were treated with varying concentrations of tunicamycin(TM)for 24h,and the expression of SIRT1 and C/EBP homologous protein(CHOP)was assessed using real-time quantitative polymerase chain reaction(RT-PCR),Western blotting,and immunofluorescence.Cell morphology and proliferation was evaluated using an inverted microscope and cell counting kit-8(CCK-8)assay,respectively.In the SRA01/04 cell apoptosis model,which underwent siRNA transfection for SIRT1 knockdown and SRT1720 treatment for its activation,the expression levels of SIRT1,CHOP,glucose regulated protein 78(GRP78),and activating transcription factor 4(ATF4)were examined.The potential reversal of SIRT1 knockdown effects by 4-phenyl butyric acid(4-PBA;an ER stress inhibitor)was investigated.In vivo,age-related cataract(ARC)rat models were induced by sodium selenite injection,and the protective role of SIRT1,activated by SRT1720 intraperitoneal injections,was evaluated through morphology observation,hematoxylin and eosin(H&E)staining,Western blotting,and RT-PCR.RESULTS:SIRT1 expression was downregulated in TMinduced SRA01/04 cells.Besides,in SRA01/04 cells,both cell apoptosis and CHOP expression increased with the rising doses of TM.ER stress was stimulated by TM,as evidenced by the increased GRP78 and ATF4 in the SRA01/04 cell apoptosis model.Inhibition of SIRT1 by siRNA knockdown increased ER stress activation,whereas SRT1720 treatment had opposite results.4-PBA partly reverse the adverse effect of SIRT1 knockdown on apoptosis.In vivo,SRT1720 attenuated the lens opacification and weakened the ER stress activation in ARC rat models.CONCLUSION:SIRT1 plays a protective role against TM-induced apoptosis in HLECs and slows the progression of cataract in rats by inhibiting ER stress.These findings suggest a novel strategy for cataract treatment focused on targeting ER stress,highlighting the therapeutic potential of SIRT1 modulation in ARC development.
基金Supported by the Training Project for Young and Middleaged Core Talents in Health System of Fujian Province(No.2016-ZQN-62)Natural Science Foundation of Fujian Province(No.2020J01652).
文摘AIM:To examine the regulatory role of microRNA-204(miR-204)on silent information regulator 1(SIRT1)and vascular endothelial growth factor(VEGF)under highglucose-induced metabolic memory in human retinal pigment epithelial(hRPE)cells.METHODS:Cells were cultured with either normal(5 mmol/L)or high D-glucose(25 mmol/L)concentrations for 8d to establish control and high-glucose groups,respectively.To induce metabolic memory,cells were cultured with 25 mmol/L D-glucose for 4d followed by culture with 5 mmol/L D-glucose for 4d.In addition,exposed in 25 mmol/L D-glucose for 4d and then transfected with 100 nmol/L miR-204 control,miR-204 inhibitor or miR-204 mimic in 5 mmol/L D-glucose for 4d.Quantitative reverse transcription-polymerase chain reaction(RT-qPCR)was used to detect miR-204 mRNA levels.SIRT1 and VEGF protein levels were assessed by immunohistochemical and Western blot.Flow cytometry was used to investigate apoptosis rate.RESULTS:It was found that high glucose promoted miR-204 and VEGF expression,and inhibited SIRT1 activity,even after the return to normal glucose culture conditions.Upregulation of miR-204 promoted apoptosis inhibiting SIRT1 and increasing VEGF expression.However,downregulation of miR-204 produced the opposite effects.CONCLUSION:The study identifies that miR-204 is the upstream target of SIRT1and VEGF,and that miR-204 can protect hRPE cells from the damage caused by metabolic memory through increasing SIRT1 and inhibiting VEGF expression.
文摘目的探讨miR-204和沉默信息调节因子1(silence information regulator 1,SIRT1)在结直肠癌组织中的表达情况及其临床价值。方法收集2018年5月至2020年6月嘉兴大学附属医院胃肠外科诊治的60例结直肠癌患者的癌组织标本和癌旁组织标本为研究对象,采用实时荧光定量聚合酶链反应检测miR-204及SIRT1的基因表达情况,Pearson分析比较miR-204与SIRT1基因的表达相关性。采用免疫组织化学SABC法检测SIRT1蛋白表达,并比较不同SIRT1蛋白表达与临床病理特征的关系。Kaplan-Meier法分析不同SIRT1蛋白表达的结直肠癌患者的生存差异。结果癌组织中miR-204基因mRNA表达显著低于癌旁组织(P<0.05),SIRT1基因mRNA表达显著高于癌旁组织(P<0.05)。癌组织中SIRT1蛋白表达阳性率显著高于癌旁组织(P<0.05)。Pearson相关分析显示miR-204与SIRT1基因mRNA在癌旁组织和癌组织中的表达均呈负相关(r=–0.647、–0.737,P<0.05)。SIRT1蛋白的表达与结直肠癌的分化水平、浸润层次、淋巴结转移与否及TNM分期相关(P<0.05),与患者的年龄、性别、肿瘤大小及肿瘤部位无关(P>0.05)。Kaplan-Meier分析显示癌组织中SIRT1阳性表达患者的生存率显著低于SIRT1阴性表达患者(χ^(2)=5.001,P=0.025)。结论结直肠癌组织中miR-204表达下调,SIRT1表达上调,二者可能通过相互影响共同促进结直肠癌的转移、侵袭,并影响患者预后。
