闫子怡,白晓琳,李卓,等.DHA虾青素双酯抑制脂肪细胞与巨噬细胞共培养体系中炎症反应[J].中国海洋药物,2021,40(5):35-41. |
DHA虾青素双酯抑制脂肪细胞与巨噬细胞共培养体系中炎症反应 |
Astaxanthin-DHA diester inhibits inflammatory response in the co-culture system of adipocytes and macrophages |
投稿时间:2020-11-05 修订日期:2021-01-07 |
DOI: |
中文关键词: DHA虾青素双酯 共培养 脂解 巨噬细胞极化 |
English Keywords:astaxanthin-DHA diester, co-culture, lipolysis, macrophage polarization |
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中文摘要: |
摘要:目的 在体外水平研究DHA虾青素双酯对脂肪组织炎症的改善作用并探究其作用机制。方法 采用Transwell小室法建立脂肪细胞与巨噬细胞共培养体系,模拟体内脂肪组织炎症状态,检测培养上清中炎症因子水平以及游离脂肪酸和甘油含量,油红O染色观察脂肪细胞脂解情况,免疫荧光表征巨噬细胞极化情况,qRT-PCR测定脂肪细胞脂解与巨噬细胞极化相关基因表达。结果 DHA虾青素双酯能显著降低共培养体系中促炎因子TNF-α(P<0.01)和MCP-1(P<0.05)的水平;抑制脂肪细胞脂解,降低培养上清中甘油含量(P<0.05);促进巨噬细胞向M2型极化,显著降低巨噬细胞内M1型标志基因IL-1β、INOS的表达(P<0.01);并且显著抑制脂肪细胞脂解相关基因TNFR(P<0.01)、JNK(P<0.01)、G0S2(P<0.05)、ATGL(P<0.01)和NF-κB信号通路关键基因MyD88、NF-κB及TNF-α(P<0.01)的表达。结论 DHA虾青素双酯可降低脂肪细胞与巨噬细胞共培养体系中炎症因子分泌,抑制脂肪细胞脂解、促进巨噬细胞M2型极化,最终抑制共培养体系中的炎症反应。 |
English Summary: |
Abstract: Objective This study is aim at investigating the ameliorative effect of astaxanthin-DHA diester (DHA-AST) on adipose tissue inflammation and exploring its mechanism in vitro. Methods Transwell chamber was used to co-culture adipocytes and macrophages to simulate the inflammatory state of adipose tissue in vivo. The levels of inflammatory factors and the content of free fatty acids (FFAs) were detected in the cultured supernatant, as well as the glycerol. Oil red O staining was used to observe the lipolysis of adipocytes, the polarization of macrophages was characterized by immunofluorescence, and qRT-PCR was performed to determine the expression of genes related to lipolysis and macrophage polarization. Results DHA-AST significantly reduced the levels of TNF-α and MCP-1 in the co-culture system, inhibited the lipolysis of adipocytes, and reduced the glycerol content in the supernatant. At the same time, DHA-AST has promoted macrophage polarization to M2-type, IL-1β and INOS, the M1-type markers, was significantly reduced in macrophages. The results of qRT-PCR indicate that the expression of lipolysis related genes are significantly inhibited (such as TNFR, JNK, G0S2 and ATGL), MyD88, NF-κB and TNF-α,the key genes of NF-κB signaling pathway were also distinctly down-regulated. Conclusion DHA-AST can reduce the secretion of inflammatory factors in the co-culture system, inhibit lipolysis of adipocytes and promote M2-type polarization of macrophages, finally alleviate the inflammatory response in the co-culture system. |
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