In vitro 와 In vivo 상에서 Candida albicans로 유도한 염증인자에 미치는 Squalene의 효과

Abstract

The purpose of this study was to evaluate the effects of SQ on the C. albicans-induced the interleukin 1β (IL-1β), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), nitric oxide (NO), Caspase-3, and nuclear factor kappa B (NF-κB) in human embryonic kidney cells (HEK 293 cells) and mice kidney. The purity of SQ was approximately 99% in vitro and in vivo experiments, each groups were treated with SQ solutions or not. They were contaminated C. albicans. The production levels of TNF-α, IL-6 and Caspase-3/CPP 32 activity in culture medium or tissue solutions were quantified using ELISA kit. The expression of mRNA IL-1β, TNF-α and IL-6 was carried out by RT-PCR. The expression of NF-κB was carried out by EMSA method. In vitro experiments, the cytotoxic effects of SQ were evaluated by the MTT analysis. Investigation has been made on both the expression of mRNA, IL-1β, TNF-α, IL-6, NF-κB and production of NO, TNF-α and IL-6, Caspase-3 activity in human embryonic kidney cells (1×105cells/well) after treatment C. albicans (3×106cells/ well) or SQ. The groups were divided as follows: Group 1: Control, Group 2: C. albicans (3×106/㎖) only, Group 3: pre treated with the SQ (0.05%/㎖) for 30 min and were incubated with C. albicans (3×106/㎖), Group 4: pre treated with the SQ (0.1%/㎖) for 30 min and were incubated with C. albicans (3×106/㎖), Group 5: pre treated with SQ (0.05%/㎖) and were incubated with C. albicans(3×106/㎖), Group 6: pre treated with SQ (0.1%/㎖) and were incubated with C. albicans (3×106/㎖). In vivo experiments, it was investigated to production of NO, TNF-α, IL-6 and Caspase-3 activity after treatment C. albicans (2×106cells/㎏) or SQ (80㎖/㎏). The groups were divided as follows: Group 1: Control, Group 2: C. albicans (2×106/㎏) only, Group 3: pre treated with the SQ (80㎖/㎏) for 7 days (one per days) and were injected with C. albicans (2×106/㎏), Group 4: pre treated with the SQ (80㎖/㎏) for 3 days (one per days) and were injected with C. albicans (2×106/㎏), Group 5: pre treated with SQ (80㎖/㎏) and were injected with C. albicans (2×106/㎏). The SQ was not observed the cytotoxicity by MTT analysis. The expression of mRNA IL-1β, TNF-α and IL-6 were highly increased by C. albicans. however, SQ was decreased the expression of mRNA IL-1β, TNF-α and IL-6. The production of NO, TNF-α, IL-6 and Caspase-3 were increased by C. albicans. SQ significantly decreased the production of C. albicans-induction by dependent on time and concentrations manners (p<0.05). The expression of NF-κB was decreased by treatment of SQ. In vivo experiments, results (p<0.05) were observed to be similar with in vitro experiments. Finally, it was found that the SQ may increase the immune response on the C. albicans induced-renal damage.