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目的:探讨λ-卡拉胶及其降解物引发THP-1单核/巨噬细胞免疫响应的结构因素及其反应机制。方法:THP-1细胞经PMA诱导分化为巨噬细胞;λ-卡拉胶及其不同分子质量的降解产物刺激THP-1细胞,采用ELISA技术检测TNF-α含量,确定引发免疫响应的λ-卡拉胶种类;采用CBA技术检测卡拉胶处理后各类免疫因子的表达;用WesternBlot检测由卡拉胶引起的免疫响应的信号通路。结果:分子质量10～40ku的卡拉胶降解产物(λ-10-40k)最易引起THP-1细胞的免疫响应,用10μg/mLλ-10-40k处理24h,可增加TNF-α的分泌量,为空白对照的2.2倍(P<0.01)。λ-10-40k可通过激活TLR4-NF-κB途径以及MAPK/ERK途径诱导THP-1细胞产生TNF-α,并通过协同效应,增加LPS刺激的TNF-α、IL-1β及IL-6的表达,放大LPS诱导的炎症反应。结论:分子质量范围10～40ku的卡拉胶降解产物易通过TLR4-NF-κB途径及MAPK/ERK途径引发THP-1单核/巨噬细胞免疫响应,并协同增加LPS刺激的炎症反应。
OBJECTIVE: To investigate the structural factors and response mechanisms of λ-carrageenan and its degradation products to induce THP-1 monocyte / macrophage immune response. METHODS: THP-1 cells were induced to differentiate into macrophages by PMA. Λ-carrageenan and its degradation products of different molecular weights stimulated THP-1 cells. The content of TNF-α was detected by ELISA and the λ-carrage The type of collagen was detected by CBA. The expression of various immune factors after carrageenan treatment was detected. Western Blot was used to detect the signal pathways of immune responses induced by carrageenan. Results: The immune response of THP-1 cells was most easily induced by carrageenan degradation product (λ-10-40k) with molecular mass of 10 ~ 40ku. The treatment with 10μg / mL λ-10-40k for 24h could increase the secretion of TNF- 2.2 times of the blank control (P <0.01). λ-10-40k can induce THP-1 cells to produce TNF-α by activating TLR4-NF-κB pathway and MAPK / ERK pathway, and through synergistic effect, increase LPS-stimulated TNF-α, IL-1β and IL-6 Expressed and amplified LPS-induced inflammatory response. CONCLUSION: The degradation products of carrageenan with mass range of 10 ~ 40 ku easily induce THP-1 monocyte / macrophage immune response through TLR4-NF-κB pathway and MAPK / ERK pathway and synergistically increase the inflammatory response induced by LPS.