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. 2020 Sep 3;20(5):45. doi: 10.3892/etm.2020.9173

Figure 5.

Figure 5

Non-cytotoxic dose of shikonin suppresses LPS-induced TNF-α expression via AMPK activation in the ex vivo cultured PBMCs from patients with COPD. After treating with the indicated doses of shikonin for 24 h, the survival and death rates of the ex vivo cultured PBMCs from patients with COPD were evaluated by (A) MTT and (B) trypan blue exclusion assays, respectively. (C) Protein expression levels of ACC, p-ACC, AMPKα and p-AMPKα were detected by western blotting. Relative ROS intensity and NF-κB activation were examined by (D) flow cytometry and (E) p65 DNA-binding activity assay, respectively. Extracellular contents and relative mRNA levels of TNF-α were examined using (F) ELISA and (G) reverse transcription-quantitative PCR, respectively. *P<0.05 vs. LPS alone group. #P<0.05 vs. corresponding shikonin combined with LPS group. C, control; LPS, lipopolysaccharide; TNF-α, tumor necrosis factor α; p-, phosphorylated; ACC, acetyl-CoA carboxylase; AMPK, AMP-activated protein kinase; IKKα/β, IκB kinase α/β; CC, Compound C; ROS, reactive oxygen species; COPD, Chronic obstructive pulmonary disease; PBMCs, peripheral blood mononuclear cells.