Anesth Pain Med.  2020 Jan;15(1):41-48. 10.17085/apm.2020.15.1.41.

Protective role of curcumin against lipopolysaccharide-induced inflammation and apoptosis in human neutrophil

• Cho KB
• Park CH
• Kim J
• Tin TD
• Kwak SH

• KMID: 2500453
• DOI: http://doi.org/10.17085/apm.2020.15.1.41

Abstract

Background
Sepsis, an uncontrolled host response to infection, may be life-threatening organ injury. Neutrophils play a critical role in regulation of host immune response to infection. Curcumin, known as a spice and food coloring agent, possesses anti-inflammatory properties. In this study, we investigated the effects of curcumin on lipopolysaccharide (LPS)-induced neutrophil activation with its signaling pathways.
Methods
Isolated human neutrophils were incubated without or with LPS and curcumin, and the expression of pro-inflammatory cytokines, such as tumor necrosis factor alpha (TNF-α), interleukin (IL)-6, and IL-8 were assessed by enzyme-linked immunosorbent assays. The expression of mitogen-activated protein kinases such as p38, extracellularsignal- regulated kinase (ERK)1/2, and c-Jun N-terminal kinase (JNK) were evaluated by Western blot analysis. Neutrophil apoptosis was also measured by fluorescenceactivated cell sorting (annexin V/propidium iodide) in LPS-stimulated neutrophils under treatment with curcumin.
Results
Curcumin attenuated expression of TNF-α, IL-6, and IL-8 and the phosphorylation levels of p38 and JNK, but not ERK1/2, in LPS-stimulated neutrophils. Additionally, curcumin restored the delayed neutrophil apoptosis by LPS-stimulated neutrophils(19.7 ± 3.2 to 38.2 ± 0.5%, P < 0.05).
Conclusions
Our results reveal the underlying mechanism of how curcumin attenuate neutrophil activation and suggest potential clinic applications of curcumin supplementation for patients with severe sepsis and septic shock. Additional clinical studies are required to confirm these in vitro findings.

Keyword

Apoptosis; Curcumin; Inflammation; Lipopolysaccharides; Neutrophils

Figure

• 

  Fig. 1 The effects of curcumin on pro-inflammatory cytokine (tumor necrosis factor alpha [TNF-α], interleukin [IL]-6, IL-8) expression in human neutrophil stimulated by lipopolysaccharide (LPS). Neutrophils (5 × 106/ml) from human blood were incubated for 4 h without or with curcumin (1, 10, 30, 50, and 100 µM), or with LPS (100 ng/ml) or LPS plus curcumin (control, curcumin, LPS, LPS + curcumin respectively). Protein results were obtained from ELISA. Data are presented as mean ± SD (n = 6 per group). *P < 0.05 vs. control. †P < 0.05 vs. LPS.

• 

  Fig. 2 The effects of curcumin on mitogen-activated protein kinases activation in human neutrophil stimulated by lipopolysaccharide (LPS). Neutrophils (5 × 106/ml) from human blood were incubated for 30 min without or with curcumin (50, 100 µM) or with LPS (100 ng/ml) or LPS plus curcumin (control, curcumin, LPS, LPS + curcumin respectively). The phosphorylation (p) and total (t) levels of p38, extracellular-signalregulated kinase (ERK)1/2, and c-Jun N-terminal kinase (JNK) were measured by Western blot analysis. Relative increase is the ratio of phosphorylated to the total kinases. Data are shown as mean ± SD (n = 4 per group). *P < 0.05 vs. control. †P < 0.05 vs. LPS.

• 

  Fig. 3 The effects of curcumin on apoptosis of human neutrophils stimulated by lipopolysaccharide (LPS). Neutrophils (5 × 106/ml) from human blood were incubated for 24 h without or with curcumin (100 µM), or with LPS (100 ng/ml) or LPS plus curcumin (control, curcumin, LPS, LPS + curcumin respectively). (A) Contour diagram of fluorescein isothiocyanate annexin V/propidium iodide (FITC-Annexin V/PI) flow cytometry of neutrophils for different groups. The lower right quadrants represent the apoptotic cells, FITC-Annexin V positive and PI negative. One representative experiment out of six is shown. (B) The percentage of neutrophil apoptosis was determined for each group. Data are shown as mean ± SD (n = 6 per group). *P < 0.05 vs. control. †P < 0.05 vs. LPS.

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