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. 2021 Jan 11;6(1):e141341. doi: 10.1172/jci.insight.141341

Figure 3. In vitro CPB conditions upregulate IL-8 and TNF-α in THP-1 cells.

Figure 3

(A) The design of in vitro CPB system. THP-1 cells at a density of 2 million cells/mL were pumped by a roller pump through PVC tubing at an average shear stress of 2.1 Pa. The majority of the tubing was kept in the water bath that maintained the temperature at 30°C. Cells were warmed up to 37°C for recovery. (B and C) The CPB conditions of high shear stress, low temperature (High S_Low T) significantly upregulated the mRNA and protein levels of IL-8 (B) and TNF-α (C) in the sheared THP-1 cells compared with static cells (n = 3 replicates/group). *P < 0.05, 1-way ANOVA and post hoc Tukey’s test. (D) Static THP-1 cells incubated with PVC tubing for 2 hours significantly upregulated IL8 by approximately 5-fold. Level of TNFA was not changed (n = 3 replicates/group). *P < 0.05, 2-tailed Student’s t test. (E and F) When THP-1 cells were sheared at 37°C (High S_High T) or at low shear stress of 0.67 Pa (Low S_Low T and Low S_High T), upregulation of mRNA and protein levels of IL-8 (E) and TNF-α (F) were significantly reduced compared with when cells were sheared at CPB conditions (High S_Low T) (n = 3–5 replicates/group). *P < 0.05, 1-way ANOVA and post hoc Dunnett’s test.