Comparison of the Regenerative Metabolic Efficiency of Lipid Extracts from Microalgae Nannochloropsis oceanica and Chlorococcum amblystomatis on Fibroblasts

Abstract

UVA radiation leads to oxidative stress and inflammation in skin cells. Therefore, the aim of this study was to compare the effect of lipid extracts from microalgae Nannochloropsis oceanica (N.o.) (marine) and Chlorococcum amblystomatis (C.a.) (freshwater) on the redox balance and PUFA metabolism in human skin fibroblasts modified by UVA. Lipid extracts from both types of microalgae introduced into the fibroblast medium after UVA irradiation significantly reduced the level of ROS and enhanced expression of Nrf2, which increased the activity/level of antioxidants (SOD1/2, CAT, GSH, Trx). The reduction in oxidative stress was accompanied by a decrease in the level of 4-HNE, its protein adducts and protein carbonyl groups. Microalgae also reduced the activity of COX1/2, FAAH and MAGL increased by UVA, and as a consequence, the level of lipid mediators (especially after N.o.) decreased, both from the group of endocannabinoids (AEA, 2-AG, PEA) and eicosanoids (PGE2, 15d-PGJ2, TXB2, 15-HETE), acting mainly through receptors related to G protein, the expression of which increases after UVA. This further contributed to the reduction in oxidative stress and pro-inflammatory signaling at NF-κB and TNFα levels. Therefore, it is suggested that lipid extracts from both N.o. and C.a. microalgae can be used to regenerate fibroblast metabolism disturbed by UVA radiation.

Keywords: fibroblasts; inflammation UV radiation; microalgae lipids extracts; phospholipid and free fatty acids metabolism; redox balance.

Figure 1

Effect of Chlorococcum amblystomatis (C.a.) or Nannochloropsis oceanica (N.o.) microalgae extract on the level of reactive oxygen species (ROS) in the following groups of fibroblasts: o control (n = 5): control cells without and after UVA irradiation [20 J/cm²]; o C.a. (n = 5): cells cultured for 24 h with C.a. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]; o N.o. (n = 5): cells cultured for 24 h with N.o. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]. Mean values ± SD and statistically significant differences for p < 0.05 are presented: a—vs. control; c—UVA+algae N.o. vs. UVA+algae C.a.; x—vs. UVA; y—UVA+algae C.a. vs. algae C.a.

Figure 2

Effect of Chlorococcum amblystomatis (C.a.) or Nannochloropsis oceanica (N.o.) microalgae extract on the expression of Nrf2 transcription factor, its main product (HO-1) and inhibitor (Keap1) in the following groups of fibroblasts: o control (n = 5): control cells without and after UVA irradiation [20 J/cm²]; o C.a. (n = 5): cells cultured for 24 h with C.a. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]; o N.o. (n = 5): cells cultured for 24 h with N.o. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]. Mean values ± SD and statistically significant differences for p < 0.05 are presented: a—vs. control; b—algae N.o. vs. algae C.a.; c—UVA+algae N.o. vs. UVA+algae C.a.; x—vs. UVA; y—UVA+algae C.a. vs. algae C.a.; z—UVA+algae N.o. vs. algae N.o.

Figure 3

Effect of Chlorococcum amblystomatis (C.a.) or Nannochloropsis oceanica (N.o.) microalgae extracts on the activity of cytosol and mitochondrial superoxide dismutases (SOD1 and SOD2) and catalase (CAT) in the following groups of fibroblasts: o control (n = 5): control cells without and after UVA irradiation [20 J/cm²]; o C.a. (n = 5): cells cultured for 24 h with C.a. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]; o N.o. (n = 5): cells cultured for 24 h with N.o. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]. Mean values ± SD and statistically significant differences for p < 0.05 are presented: a—vs. control; c—UVA+algae N.o. vs. UVA+algae C.a.; x—vs. UVA; y—UVA+algae C.a. vs. algae C.a.; z—UVA+algae N.o. vs. algae N.o.

Figure 4

Effect of Chlorococcum amblystomatis (C.a.) or Nannochloropsis oceanica (N.o.) microalgae extracts on the elements of glutathione and thioredoxin systems in the following groups of fibroblasts: o control (n = 5): control cells without and after UVA irradiation [20 J/cm²]; o C.a. (n = 5): cells cultured for 24 h with C.a. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]; o N.o. (n = 5): cells cultured for 24 h with N.o. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]. Mean values ± SD and statistically significant differences for p < 0.05 are presented: a—vs. control; b—algae N.o. vs. algae C.a.; c—UVA+algae N.o. vs. UVA+algae C.a.; x—vs. UVA; y—UVA+algae C.a. vs. algae C.a.; z—UVA+algae N.o. vs. algae N.o.

Figure 5

Effect of Chlorococcum amblystomatis (C.a.) or Nannochloropsis oceanica (N.o.) microalgae extracts on the level of 4-HNE, 4-HNE-protein adducts and protein carboxyl groups (CBO) in the following fibroblast groups: o control (n = 5): control cells without and after UVA irradiation [20 J/cm²]; o C.a. (n = 5): cells cultured for 24 h with C.a. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]; o N.o. (n = 5): cells cultured for 24 h with N.o. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]. Mean values ± SD and statistically significant differences for p < 0.05 are presented: a—vs. control; b—algae N.o. vs. algae C.a.; c—UVA+algae N.o. vs. UVA+algae C.a.; x—vs. UVA; y—UVA+algae C.a. vs. algae C.a.; z—UVA+algae N.o. vs. algae N.o.

Figure 6

Effect of Chlorococcum amblystomatis (C.a.) or Nannochloropsis oceanica (N.o.) microalgae extracts on the level of free and phospholipid (PL) non-saturated fatty acids in the following groups of fibroblasts: o control (n = 5): control cells without and after UVA irradiation [20 J/cm²]; o C.a. (n = 5): cells cultured for 24 h with C.a. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]; o N.o. (n = 5): cells cultured for 24 h with N.o. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]. Mean values ± SD and statistically significant differences for p < 0.05 are presented: a—vs. control; b—algae N.o. vs. algae C.a.; c—UVA+algae N.o. vs. UVA+algae C.a.; x—vs. UVA; y—UVA+algae C.a. vs. algae C.a.; z—UVA+algae N.o. vs. algae N.o.

Figure 7

Effect of Chlorococcum amblystomatis (C.a.) or Nannochloropsis oceanica (N.o.) microalgae extracts on the activity of cyclooxygenase-1/2, cytosolic phospholipase A2 and 5-lipoxygenase (5-LOX) in the following fibroblast groups: o control (n = 5): control cells without and after UVA irradiation [20 J/cm²]; o C.a. (n = 5): cells cultured for 24 h with C.a. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]; o N.o. (n = 5): cells cultured for 24 h with N.o. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]. Mean values ± SD and statistically significant differences for p < 0.05 are presented: a—vs. control; b—algae N.o. vs. algae C.a.; c—UVA+algae N.o. vs. UVA+algae C.a.; x—vs. UVA; y—UVA+algae C.a. vs. algae C.a.; z—UVA+algae N.o. vs. algae N.o.

Figure 8

Effect of Chlorococcum amblystomatis (C.a.) or Nannochloropsis oceanica (N.o.) microalgae extracts on the level of eicosanoids in the following fibroblast groups: o control (n = 5): control cells without and after UVA irradiation [20 J/cm²]; o C.a. (n = 5): cells cultured for 24 h with C.a. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]; o N.o. (n = 5): cells cultured for 24 h with N.o. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]. Mean values ± SD and statistically significant differences for p < 0.05 are presented: a—vs. control; b—algae N.o. vs. algae C.a.; c—UVA+algae N.o. vs. UVA+algae C.a.; x—vs. UVA; y—UVA+algae C.a. vs. algae C.a.; z—UVA+algae N.o. vs. algae N.o.

Figure 9

Effect of Chlorococcum amblystomatis (C.a.) or Nannochloropsis oceanica (N.o.) microalgae extracts on the level of endocannabinoids (anandamide (AEA), 2-arachidonylglycerol (2-AG) and derivative compounds—palmitoylethanolamide (PEA) and oleoylethanolamide (OEA)) and the activity of AEA and 2-AG metabolizing enzymes (FAAH and MAGL) in the following fibroblast groups: o control (n = 5): control cells without and after UVA irradiation [20 J/cm²]; o C.a. (n = 5): cells cultured for 24 h with C.a. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]; o N.o. (n = 5): cells cultured for 24 h with N.o. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]. Mean values ± SD and statistically significant differences for p < 0.05 are presented: a—vs. control; b—algae N.o. vs. algae C.a.; c—UVA+algae N.o. vs. UVA+algae C.a.; x—vs. UVA; y—UVA+algae C.a. vs. algae C.a.; z—UVA+algae N.o. vs. algae N.o.

Figure 10

Effect of Chlorococcum amblystomatis (C.a.) or Nannochloropsis oceanica (N.o.) microalgae extracts on the expression of G-protein-coupled receptors (CB1, CB2, PPARγ, TRPV1) in the following fibroblast groups: o control (n = 5): control cells without and after UVA irradiation [20 J/cm²]; o o C.a. (n = 5): cells cultured for 24 h with C.a. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]; o N.o. (n = 5): cells cultured for 24 h with N.o. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]. Mean values ± SD and statistically significant differences for p < 0.05 are presented: a—vs. control; b—algae N.o. vs. algae C.a.; c—UVA+algae N.o. vs. UVA+algae C.a.; x—vs. UVA; y—UVA+algae C.a. vs. algae C.a.; z—UVA+algae N.o. vs. algae N.o.

Figure 11

Effect of Chlorococcum amblystomatis (C.a.) or Nannochloropsis oceanica (N.o.) microalgae extracts on the level of subunits of transcription factor NF-κB (p52 and p65) and cytokine TNFα in the following fibroblast groups: o control (n = 5): control cells without and after UVA irradiation [20 J/cm²]; o o C.a. (n = 5): cells cultured for 24 h with C.a. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]; o N.o. (n = 5): cells cultured for 24 h with N.o. [2 μg/mL] without irradiation and after irradiation with UVA [20 J/cm²]. Mean values ± SD and statistically significant differences for p < 0.05 are presented: a—vs. control; c—UVA+algae N.o. vs. UVA+algae C.a.; x—vs. UVA; y—UVA+algae C.a. vs. algae C.a.; z—UVA+algae N.o. vs. algae N.o.