【摘 要】
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Objective:To test the hypothesis that the inhibition of endoplasmic reticulum (ER) stress-induced apoptosis in oxidized low-density lipoproteins (ox-LDL)-induced human aortic-vascular smooth muscle cells (HA-VSMCs) was associated with suppression of the p
【机 构】
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Cardiovascular Department, Shenzhen Traditional Chinese Medicine Hospital, Guangzhou University of T
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Objective:To test the hypothesis that the inhibition of endoplasmic reticulum (ER) stress-induced apoptosis in oxidized low-density lipoproteins (ox-LDL)-induced human aortic-vascular smooth muscle cells (HA-VSMCs) was associated with suppression of the protein kinase RNA-like ER kinase (PERK)-eukaryotic translation initiation factor 2α (elF2α)-activating transcription factor 4 (ATF4)-CCAAT/enhancer binding protein homologous protein (CHOP) signaling pathway by Pollen Typhae total flavone (PTF).Methods:Primany HA-VSMCs were cultured and identified.The cultured HA-VSMCs were randomized into 5 groups,including a normal control group,an ox-LDL group (70 μ g/mL high ox-LDL),an HPTF group (70 μ g/mL high ox-LDL+500 μ g/mL PTF),an MPTF group (70 μg/mL high ox-LDL+250 μg/mL PTF),and a LPTF group (70 μg/mL high ox-LDL+100 μg/mL PTF) in the first part;and a normal control group,an ox-LDL group (70 μ g/mL high ox-LDL),an MPTF group (70 μg/mL high ox-LDL+250 μg/mL PTF),a shRNA group (transducted with PERK shRNA lentiviral particles),a scramble shRNA group (transducted with control shRNA lentiviral particles),an MPTF+ox-LDL+shRNA group (250 μg/mL PTF+70 μ g/mL high ox-LDL+PERK shRNA lentiviral particles) and an ox-LDL+shRNA group (70 μ g/mL high ox-LDL+PERK shRNA lentiviral particles) in the second part.The protein expression levels of ER-associated apoptosis proteins were detected by Westem blot,and their mRNA expression levels were detected by quantitative real-time reverse transcription-polymerase chain reaction.The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was applied to test cell viability,and the level of apoptosis was monitored by flow cytometry.Results:The MTT assay and flow cytometry showed that the ox-LDL group had a significant increase in apoptosis,which was attenuated in PTF treatment groups and shRNA groups.Moreover,the ox-LDL group had increased protein and mRNA levels of binding immunoglobulin protein and ER-associated apoptosis proteins,such as PERK,elF2α,ATF4 and CHOP,which were attenuated in PTF treatment groups and shRNA groups.Conclusions:The apoptosis induced by ox-LDL had a strong relation to ER stress.The protective effect of PTF on ER stress-induced apoptosis was associated with inhibition of the PERK-elF2α-ATF4-CHOP pathway,which might be a potential therapeutic strategy for enhancing the stability of atherosclerotic plaques.
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