目的:探究虎杖苷调控微小RNA(microRNA,miR)-128-3p/SIRT/自噬促进糖尿病(DM)溃疡模型大鼠创面愈合的机制。方法:通过腹腔注射链脲佐菌素(70 mg/kg)建立DM模型。将30只DM大鼠随机分为模型组(n=15)、虎杖苷组(n=15)。另取15只健康大鼠作...目的:探究虎杖苷调控微小RNA(microRNA,miR)-128-3p/SIRT/自噬促进糖尿病(DM)溃疡模型大鼠创面愈合的机制。方法:通过腹腔注射链脲佐菌素(70 mg/kg)建立DM模型。将30只DM大鼠随机分为模型组(n=15)、虎杖苷组(n=15)。另取15只健康大鼠作为对照组。3组均通过去皮建立DM伤口损伤模型。虎杖苷组大鼠灌胃虎杖苷(20 mg/kg)。比较各组大鼠伤口愈合情况、愈合周围组织中超氧化物歧化酶(SOD)、丙二醛(MDA)和自噬标志蛋白表达水平。分析miR-128-3p和SIRT1的表达水平。通过双荧光素酶报告在内皮祖细胞中验证miR-128-3p与SIRT1的靶向关系。通过转染miR-128-3p mimic质粒过表达miR-128-3p。结果:第5天,模型组、虎杖苷组大鼠伤口愈合率低于对照组(P<0.05);第10天,虎杖苷组大鼠伤口愈合率高于模型组(P<0.05),低于对照组(P<0.05)。模型组大鼠创面组织中SOD、SIRT1 m RNA相对表达量、SIRT1蛋白相对表达量、LC3Ⅱ蛋白相对表达量、Beclin1蛋白相对表达量、LC3Ⅱ/Ⅰ均低于对照组(P<0.05),而MDA和miR-128-3p水平显著高于对照组(P<0.05)。虎杖苷组大鼠创面组织中SOD、SIRT1 m RNA相对表达量、SIRT1蛋白相对表达量、LC3Ⅱ蛋白相对表达量、Beclin1蛋白相对表达量、LC3Ⅱ/Ⅰ均高于模型组(P<0.05),而MDA、miR-128-3p均低于模型组(P<0.05)。双荧光素酶报告结果显示miR-128-3p可以与SIRT1靶向结合。转染miR-128-3p mimic后,内皮祖细胞中的miR-128-3p的水平显著升高,并且SIRT1 m RNA和SIRT1蛋白表达水平显著降低。结论:虎杖苷可能通过调控miR-128-3p/SIRT1通路诱导自噬,进而缓解高血糖引起的氧化应激损伤,促进DM模型大鼠的伤口愈合。展开更多
Background:Long-chain non-coding RNA(lncRNA)LINC00609 is a potential tumor suppressor,but the mechanism of action in non-small cell lung cancer(NSCLC)is yet to be understood.Objectives:The effects of LINC00609 on A549...Background:Long-chain non-coding RNA(lncRNA)LINC00609 is a potential tumor suppressor,but the mechanism of action in non-small cell lung cancer(NSCLC)is yet to be understood.Objectives:The effects of LINC00609 on A549 cell proliferation,apoptosis,and cell cycle arrest were investigated.Methods:The LINC00609 levels in NSCLC and normal tissues were analyzed by bioinformatics.Expressions of LINC00609,miR-128-3p,and Rho family GTPase 3(RND3)in NSCLC cells(A549)were determined by qRT-PCR.Bioinformatics analysis predicted target genes and dual-luciferase reporter assays to ensure that LINC00609 targeted miR-128-3p and miR-128-3p targeted RND3.The proliferation of cells was determined using EDU and CCK-8.Flow cytometry was used to evaluate cell apoptosis rate and cell cycle.The western blotting assay identified proteins related to proliferation and apoptosis.Results:In NSCLC tissues,LINC00609 was expressed in low levels,while its high expression was associated with a higher survival rate.LINC00609 affected cell proliferation,apoptosis,cell cycle arrest,and expression of related proteins.Dual-luciferase reporter assay showed that LINC00609 binds specifically to miR-128-3p,and miR-128-3p binds to RND3.MiR-128-3p overexpression could neutralize the effects of LINC00609.A siRNA targeting RND3 could reverse the effect of the miR-128-3p inhibitor.Silencing RND3 resulted in a decrease in apoptosis rate and the number of cells in the S-phase and an increase in the number of cells in the G1-phase.Furthermore,phosphorylation levels of the AKT protein and mTOR protein,and Bcl2 expression,increased;however,the expression of RND3,Bax,and caspase3 decreased.Conclusions:LINC00609 regulated miR-128-3p/RND3 axis to modulate A549 cell proliferation,apoptosis,and cell cycle arrest.In the case of NSCLC,LINC00609 could be a potential target for therapy.展开更多
文摘目的:探究虎杖苷调控微小RNA(microRNA,miR)-128-3p/SIRT/自噬促进糖尿病(DM)溃疡模型大鼠创面愈合的机制。方法:通过腹腔注射链脲佐菌素(70 mg/kg)建立DM模型。将30只DM大鼠随机分为模型组(n=15)、虎杖苷组(n=15)。另取15只健康大鼠作为对照组。3组均通过去皮建立DM伤口损伤模型。虎杖苷组大鼠灌胃虎杖苷(20 mg/kg)。比较各组大鼠伤口愈合情况、愈合周围组织中超氧化物歧化酶(SOD)、丙二醛(MDA)和自噬标志蛋白表达水平。分析miR-128-3p和SIRT1的表达水平。通过双荧光素酶报告在内皮祖细胞中验证miR-128-3p与SIRT1的靶向关系。通过转染miR-128-3p mimic质粒过表达miR-128-3p。结果:第5天,模型组、虎杖苷组大鼠伤口愈合率低于对照组(P<0.05);第10天,虎杖苷组大鼠伤口愈合率高于模型组(P<0.05),低于对照组(P<0.05)。模型组大鼠创面组织中SOD、SIRT1 m RNA相对表达量、SIRT1蛋白相对表达量、LC3Ⅱ蛋白相对表达量、Beclin1蛋白相对表达量、LC3Ⅱ/Ⅰ均低于对照组(P<0.05),而MDA和miR-128-3p水平显著高于对照组(P<0.05)。虎杖苷组大鼠创面组织中SOD、SIRT1 m RNA相对表达量、SIRT1蛋白相对表达量、LC3Ⅱ蛋白相对表达量、Beclin1蛋白相对表达量、LC3Ⅱ/Ⅰ均高于模型组(P<0.05),而MDA、miR-128-3p均低于模型组(P<0.05)。双荧光素酶报告结果显示miR-128-3p可以与SIRT1靶向结合。转染miR-128-3p mimic后,内皮祖细胞中的miR-128-3p的水平显著升高,并且SIRT1 m RNA和SIRT1蛋白表达水平显著降低。结论:虎杖苷可能通过调控miR-128-3p/SIRT1通路诱导自噬,进而缓解高血糖引起的氧化应激损伤,促进DM模型大鼠的伤口愈合。
基金supported by the Fundamental Research Funds for the Central Universities(No.2042021kf1038).
文摘Background:Long-chain non-coding RNA(lncRNA)LINC00609 is a potential tumor suppressor,but the mechanism of action in non-small cell lung cancer(NSCLC)is yet to be understood.Objectives:The effects of LINC00609 on A549 cell proliferation,apoptosis,and cell cycle arrest were investigated.Methods:The LINC00609 levels in NSCLC and normal tissues were analyzed by bioinformatics.Expressions of LINC00609,miR-128-3p,and Rho family GTPase 3(RND3)in NSCLC cells(A549)were determined by qRT-PCR.Bioinformatics analysis predicted target genes and dual-luciferase reporter assays to ensure that LINC00609 targeted miR-128-3p and miR-128-3p targeted RND3.The proliferation of cells was determined using EDU and CCK-8.Flow cytometry was used to evaluate cell apoptosis rate and cell cycle.The western blotting assay identified proteins related to proliferation and apoptosis.Results:In NSCLC tissues,LINC00609 was expressed in low levels,while its high expression was associated with a higher survival rate.LINC00609 affected cell proliferation,apoptosis,cell cycle arrest,and expression of related proteins.Dual-luciferase reporter assay showed that LINC00609 binds specifically to miR-128-3p,and miR-128-3p binds to RND3.MiR-128-3p overexpression could neutralize the effects of LINC00609.A siRNA targeting RND3 could reverse the effect of the miR-128-3p inhibitor.Silencing RND3 resulted in a decrease in apoptosis rate and the number of cells in the S-phase and an increase in the number of cells in the G1-phase.Furthermore,phosphorylation levels of the AKT protein and mTOR protein,and Bcl2 expression,increased;however,the expression of RND3,Bax,and caspase3 decreased.Conclusions:LINC00609 regulated miR-128-3p/RND3 axis to modulate A549 cell proliferation,apoptosis,and cell cycle arrest.In the case of NSCLC,LINC00609 could be a potential target for therapy.
