BACKGROUND Plantamajoside(PMS)has shown potential in mitigating cell damage caused by high glucose(HG)levels.Despite this,the precise therapeutic effects of PMS on type 2 diabetes mellitus(T2DM)and the underlying regu...BACKGROUND Plantamajoside(PMS)has shown potential in mitigating cell damage caused by high glucose(HG)levels.Despite this,the precise therapeutic effects of PMS on type 2 diabetes mellitus(T2DM)and the underlying regulatory mechanisms require further exploration.AIM To investigate PMS therapeutic effects on T2DM in mice and elucidate its mechanisms of action through in vivo and in vitro experiments.METHODS An in vitro damage model of MIN6 cells was established using HG and palmitic acid(PA).PMS's protective effect on cell damage was assessed.Next,transcriptomics was employed to examine how PMS treatment affects gene expression of MIN6 cells.Furthermore,the effect of PMS on protein processing in endoplasmic reticulum and apoptosis pathways was validated.A T2DM mouse model was used to validate the therapeutic effects and mechanisms of PMS in vivo.RESULTS PMS intervention ameliorated cell injury in HG+PA-induced MIN6 cell damage.Transcriptomic analysis revealed that protein processing in the endoplasmic reticulum and apoptosis pathways were enriched in cells treated with PMS,with significant downregulation of the gene Dnajc1.Further validation indicated that PMS significantly inhibited the expression of apoptosis-related factors(Bax,CytC)and endoplasmic reticulum stress(ERS)-related factors[ATF6,XBP1,Ddit3(CHOP),GRP78],while promoting the expression of Bcl-2 and Dnajc1.Additionally,the inhibitory effects of PMS on ERS and apoptosis were abolished upon Dnajc1 silencing.Furthermore,in vivo experiments demonstrated that PMS intervention effectively improved pancreatic damage,suppressed the expression of apoptosis-related factors(Bax,CytC),and ERS-related factors[ATF6,XBP1,Ddit3(CHOP),GRP78],while promoting the expression of Bcl-2 and Dnajc1 in a T2DM model mice.CONCLUSION PMS intervention could alleviate pancreatic tissue damage effectively.The mechanism of action involves Dnajc1 activation,which subsequently inhibits apoptosis and ERS,ameliorating damage to pancreaticβ-cells.展开更多
目的探讨LIGHT(lymphotoxin like,exhibits inducible expression and competes with HSV glycoprotein D for HVEM,a receptor expressed by T lymphocytes)协同γ-干扰素(interferon-gamma,IFN-γ)诱导小鼠胰岛β细胞株MIN6细胞凋亡...目的探讨LIGHT(lymphotoxin like,exhibits inducible expression and competes with HSV glycoprotein D for HVEM,a receptor expressed by T lymphocytes)协同γ-干扰素(interferon-gamma,IFN-γ)诱导小鼠胰岛β细胞株MIN6细胞凋亡的作用及机制。方法 LIGHT、IFN-γ单独或联合处理MIN6细胞。分别运用CCK-8法、流式细胞术检测MIN6细胞增殖与凋亡率。Western blot检测信号转导和转录激活子1(signal transducers and activators of transcription 1,STAT1)及STAT1的磷酸化水平,以及凋亡与增殖相关基因Bcl-xL、Bax和增殖细胞核抗原(PCNA)的表达。MIN6细胞经STAT1特异的抑制剂弗达拉滨(fludarabine,Flu)预处理后再用LIGHT和IFN-γ联合处理,Western blot检测STAT1、p-STAT1、Bcl-xL、Bax的变化,CCK-8法检测MIN6细胞的增殖能力。结果 LIGHT协同IFN-γ呈时间依赖性抑制MIN6细胞增殖,诱导细胞凋亡,降低抗凋亡基因Bcl-xL表达、增加促凋亡基因Bax表达,对PCNA表达无明显影响;LIGHT促进IFN-γ激活STAT1,而STAT1抑制剂fludarabine抑制STAT1活化后,Bcl-xL表达上调,Bax表达下调,MIN6细胞增殖率上升。结论 LIGHT协同IFN-γ通过激活STAT1调节Bcl-xL、Bax表达而诱导小鼠胰岛β细胞株MIN6细胞凋亡。展开更多
基金Yuansong Wang National Famous Traditional Chinese Medicine Expert Heritage Studio,No.4(2022).
文摘BACKGROUND Plantamajoside(PMS)has shown potential in mitigating cell damage caused by high glucose(HG)levels.Despite this,the precise therapeutic effects of PMS on type 2 diabetes mellitus(T2DM)and the underlying regulatory mechanisms require further exploration.AIM To investigate PMS therapeutic effects on T2DM in mice and elucidate its mechanisms of action through in vivo and in vitro experiments.METHODS An in vitro damage model of MIN6 cells was established using HG and palmitic acid(PA).PMS's protective effect on cell damage was assessed.Next,transcriptomics was employed to examine how PMS treatment affects gene expression of MIN6 cells.Furthermore,the effect of PMS on protein processing in endoplasmic reticulum and apoptosis pathways was validated.A T2DM mouse model was used to validate the therapeutic effects and mechanisms of PMS in vivo.RESULTS PMS intervention ameliorated cell injury in HG+PA-induced MIN6 cell damage.Transcriptomic analysis revealed that protein processing in the endoplasmic reticulum and apoptosis pathways were enriched in cells treated with PMS,with significant downregulation of the gene Dnajc1.Further validation indicated that PMS significantly inhibited the expression of apoptosis-related factors(Bax,CytC)and endoplasmic reticulum stress(ERS)-related factors[ATF6,XBP1,Ddit3(CHOP),GRP78],while promoting the expression of Bcl-2 and Dnajc1.Additionally,the inhibitory effects of PMS on ERS and apoptosis were abolished upon Dnajc1 silencing.Furthermore,in vivo experiments demonstrated that PMS intervention effectively improved pancreatic damage,suppressed the expression of apoptosis-related factors(Bax,CytC),and ERS-related factors[ATF6,XBP1,Ddit3(CHOP),GRP78],while promoting the expression of Bcl-2 and Dnajc1 in a T2DM model mice.CONCLUSION PMS intervention could alleviate pancreatic tissue damage effectively.The mechanism of action involves Dnajc1 activation,which subsequently inhibits apoptosis and ERS,ameliorating damage to pancreaticβ-cells.
文摘目的探讨LIGHT(lymphotoxin like,exhibits inducible expression and competes with HSV glycoprotein D for HVEM,a receptor expressed by T lymphocytes)协同γ-干扰素(interferon-gamma,IFN-γ)诱导小鼠胰岛β细胞株MIN6细胞凋亡的作用及机制。方法 LIGHT、IFN-γ单独或联合处理MIN6细胞。分别运用CCK-8法、流式细胞术检测MIN6细胞增殖与凋亡率。Western blot检测信号转导和转录激活子1(signal transducers and activators of transcription 1,STAT1)及STAT1的磷酸化水平,以及凋亡与增殖相关基因Bcl-xL、Bax和增殖细胞核抗原(PCNA)的表达。MIN6细胞经STAT1特异的抑制剂弗达拉滨(fludarabine,Flu)预处理后再用LIGHT和IFN-γ联合处理,Western blot检测STAT1、p-STAT1、Bcl-xL、Bax的变化,CCK-8法检测MIN6细胞的增殖能力。结果 LIGHT协同IFN-γ呈时间依赖性抑制MIN6细胞增殖,诱导细胞凋亡,降低抗凋亡基因Bcl-xL表达、增加促凋亡基因Bax表达,对PCNA表达无明显影响;LIGHT促进IFN-γ激活STAT1,而STAT1抑制剂fludarabine抑制STAT1活化后,Bcl-xL表达上调,Bax表达下调,MIN6细胞增殖率上升。结论 LIGHT协同IFN-γ通过激活STAT1调节Bcl-xL、Bax表达而诱导小鼠胰岛β细胞株MIN6细胞凋亡。
