Antibody to oxidized low-density lipoprotein inhibits THP1 cells from apoptosis by suppressing NF-κB pathway activation
Rui Dai, Jing Dong, Weijuan Li, Yi Zhou, Wei Zhou, Wenping Zhou, Manhua Chen
Abstract
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Rui Dai, Jing Dong, Weijuan Li, Yi Zhou, Wei Zhou, Wenping Zhou, Manhua Chen
Abstract
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BACKGROUND: This study aimed to investigate whether the oxidized low-density lipoprotein (Ox-LDL) antibody is able to inhibit THP1 cell apoptosis by suppressing NF-κB pathway. METHODS: THP1 cells were induced to macrophages with phorbol 12-myristate 13-acetate (PMA). Macrophages were divided into control group, Ox-LDL group and antibody group, cells in which were treated with phosphate buffered saline (PBS), Ox-LDL (50 mg/mL), Ox-LDL (50 µg/mL) plus Ox-LDL antibody (100 mg/L), respectively, for 24 h. The apoptosis rate was determined by inverted microscopy and flow cytometry. The protein and mRNA expression of NF-κB (P65), caspase-3 and BCL2 was detected by Western blotting and reverse transcription polymerase chain reaction (RT-PCR), respectively. RESULTS: The apoptosis rate reduced significantly in antibody group as compared to Ox-LDL and control groups (P<0.05). The protein and mRNA expression of NF-κB pathway was markedly lowered in antibody group than in Ox-LDL and control groups (P<0.05), which reduced the Ox-LDL induced inflammation. CONCLUSIONS: Ox-LDL antibody may be used to attenuate Ox-LDL induced inflammation and apoptosis, preventing atherosclerosis patients from acute coronary syndrome (ACS).
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BACKGROUND: This study aimed to investigate whether the oxidized low-density lipoprotein (Ox-LDL) antibody is able to inhibit THP1 cell apoptosis by suppressing NF-κB pathway. METHODS: THP1 cells were induced to macrophages with phorbol 12-myristate 13-acetate (PMA). Macrophages were divided into control group, Ox-LDL group and antibody group, cells in which were treated with phosphate buffered saline (PBS), Ox-LDL (50 mg/mL), Ox-LDL (50 µg/mL) plus Ox-LDL antibody (100 mg/L), respectively, for 24 h. The apoptosis rate was determined by inverted microscopy and flow cytometry. The protein and mRNA expression of NF-κB (P65), caspase-3 and BCL2 was detected by Western blotting and reverse transcription polymerase chain reaction (RT-PCR), respectively. RESULTS: The apoptosis rate reduced significantly in antibody group as compared to Ox-LDL and control groups (P<0.05). The protein and mRNA expression of NF-κB pathway was markedly lowered in antibody group than in Ox-LDL and control groups (P<0.05), which reduced the Ox-LDL induced inflammation. CONCLUSIONS: Ox-LDL antibody may be used to attenuate Ox-LDL induced inflammation and apoptosis, preventing atherosclerosis patients from acute coronary syndrome (ACS).
Key concepts: Apoptosis, Antibody, Flow cytometry, Molecular biology, Lipoprotein, Inflammation, Medicine, Blot