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Pollen Typhae Total Flavone Inhibits Endoplasmic Reticulum Stress-Induced Apoptosis in Human Aortic-Vascular Smooth Muscle Cells through Down-Regulating PERK-eIF2α-ATF4-CHOP Pathway

引用

Chinese Journal of Integrative Medicine 2019年第8期25卷 604-612页

作者： CHEN Ming-tai HUANG Ruo-lan OU Li-jun CHEN Ying-nan MEN Ling CHANG Xiao WANG Ling YANG Yu-zhu ZHANG Zhong Cardiovascular Department Shenzhen Traditional Chinese Medicine Hospital Guangzhou University of Traditional Chinese Medicine

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(e IF2α)-activating transcription factor 4(ATF4)-CCAAT/enhancer binding protein homologous protein(CHOP) signaling pathway by Pollen Typhae total flavone(PTF). Methods: Primary 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 μg/m L high ox-LDL), an HPTF group(70 μg/m L high ox-LDL+500 μg/m L PTF), an MPTF group(70 μg/m L high ox-LDL+250 μg/m L PTF), and a LPTF group(70 μg/m L high ox-LDL+100 μg/m L PTF) in the first part;and a normal control group, an ox-LDL group(70 μg/mL high ox-LDL), an MPTF group(70 μg/m L high ox-LDL+250 μg/m L PTF), a sh rna 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 μg/mL high ox-LDL+PERK shrna lentiviral particles) and an ox-LDL+shrna group(70 μg/mL high ox-LDL+PERK shrna lentiviral particles) in the second part. The protein expression levels of ER-associated apoptosis proteins were detected by Western blot, and their m rna 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 cel 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 sh rna groups. Moreover, the ox-LDL group had increased protein and m rna levels of binding immunoglobulin protein and ER-associated

关键词： Pollen Typhae total flavone endoplasmic reticulum stress protein kinase rna-like endoplasmic reticulum kinase-eukaryotic translation initiation factor 2a -activating transcription factor 4-CCAAT/enhancer binding protein homologous protein pathway apoptosis vulnerable atherosclerotic plaque

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