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烹调油烟急性暴露促发脂肪组织炎症反应的分子机制OA北大核心CSTPCD

Molecular mechanism of adipose tissue inflammation induced by acute exposure to cooking oil fumes

中文摘要英文摘要

[背景]烹调油烟与机体免疫反应密切相关,而脂肪组织也在免疫调节中发挥重要作用.目前关于油烟暴露诱导脂肪组织炎症的生物学效应及其作用机制尚未阐明. [目的]通过烹调油烟急性暴露,探讨油烟不同暴露时长对小鼠脂肪组织的炎症损伤效应并探索Nod样受体蛋白-3(NLRP3)/半胱天冬酶1(Caspase 1)/白介素(IL)-1β信号通路在其中的作用. [方法]将 40只 8周龄雌性C57BL/6J小鼠随机分为 4组,分别是 3 d对照组、7 d对照组、3 d暴露组和 7 d暴露组,每组 10只.采用烹调油烟动物暴露系统(COFFEE)对小鼠进行油烟吸入暴露,暴露20 min后暂停10 min,每天1 h,连续暴露3 d和7 d.每天观察小鼠一般状态并测量体重,暴露结束后眼球取血,采用酶联免疫吸附试验(ELISA)检测血清中IL-6、IL-27和IL-1β水平.采集小鼠脂肪组织,苏木精-伊红(HE)染色进行病理形态学观察,流式细胞术检测脂肪组织中CD4+和CD8+T细胞百分比,实时定量PCR(RT-qPCR)检测脂肪组织中核因子-κB(NF-κB)、NLRP3、Caspase 1和IL-1β基因表达水平,蛋白免疫印迹实验(Western blot)检测脂肪中NLRP3、Caspase 1和IL-1β蛋白表达水平. [结果]与对照组相比,3 d暴露组和 7 d暴露组小鼠的血清IL-6、IL-27和IL-1β含量均升高,除3 d暴露组IL-6外,其他差异均有统计学意义(P<0.05),且在 7 d暴露组的水平明显高于3 d暴露组(P<0.05);HE染色结果显示,7 d暴露组脂肪细胞内空泡脂滴减小,胞质皱缩,且有炎症细胞浸润;流式细胞术结果显示,3 d暴露组和 7 d暴露组小鼠脂肪细胞内CD4+和CD8+T细胞比例较对照组均有升高趋势,7 d暴露组差异有统计学意义(P<0.05),且两组的CD4+/CD8+T值也递级升高,差异有统计学意义(P<0.05);RT-qPCR结果显示,与对照组相比,3 d暴露组和 7 d暴露组的小鼠脂肪组织中NF-κB、NLRP3、Caspase 1和IL-1β mRNA表达水平均升高(P<0.05,P<0.01),7 d内各暴露组小鼠mRNA表达水平随暴露时长的增加呈现逐渐升高的趋势;Western blot结果显示,与对照组相比,3 d暴露组小鼠NLRP3和Caspase 1蛋白表达量明显升高(P<0.01),IL-1β蛋白表达有升高趋势,但差异尚无统计学意义(P>0.05),7 d暴露组的小鼠NLRP3、Caspase 1和IL-1β的蛋白表达量均高于 7 d暴露组(P<0.05,P<0.01). [结论]烹调油烟急性暴露能导致脂肪组织出现明显的炎症反应,且随着暴露时间的延长效应逐渐增强,其作用机制可能与NLRP3炎症小体信号通路激活相关.

[Background]Cooking oil fumes are closely related to immune response,and adipose tissue also plays an important role in immune regulation.At present,the biological effect and mechanism of inflammation of adipose tissue induced by oil fume exposure are not clear yet. [Objective]To investigate the inflammatory effect of different exposure duration of cooking fumes on adipose tissue in mice and explore the role of Nod-like receptor pyrin domain 3(NLRP3)/cysteinyl aspartate specific proteinase 1(Caspase 1)/interleukin(IL)-1β signaling pathway. [Methods]Forty 8-week-old female C57BL/6J mice were randomly divided into 3-day control group(CON3 group),7-day control group(CON7 group),3-day oil fume exposure group(COF3 group),and 7-day oil fume exposure group(COF7 group),with 10 mice in each group.The mice were exposed to oil fumes in a cooking oil fume formation and exposure equipment(COFFEE)for 20 min,followed by a 10-min pause,1 h a day for consecutive 3 d or 7 d.General condition of mice was observed and body weight was measured every day.After ex-posure,blood was sampled from the eyeball.Serum levels of IL-6,IL-27,and IL-1β were detected by enzyme-linked immunosorbent assay(ELISA).The adipose tissue of mice was collected and observed after hematoxylin-eosin(HE)staining.The percentages of CD4+ and CD8+T cells in adipose tissue were detected by flow cytometry.Real-time quantitative PCR(RT-qPCR)was used to detect the expression levels of nuclear factor-κB(NF-κB),NLRP3,Caspase 1,and IL-1β in adipose tissue.Western blot was used to detect the expression levels of NLRP3,Caspase 1,and IL-1β in adipose. [Results]Compared with the corresponding control group,serum IL-6,IL-27,and IL-1β contents in the COF3 group and the COF7 group were significantly increased(P<0.05)except IL-6 in the COF3 group,and the levels in the COF7 group were significantly higher than those in the COF3 group(P<0.05).Vacuolar lipid droplets in adipocytes decreased,cytoplasm shrank,and inflammatory cells infiltrated in the COF7 group after HE staining.The flow cytometry results showed that the proportions of CD4+ and CD8+T cells in adipocytes of the COF3 group and the COF7 group were increased compared to the corresponding control group,with a significant increase in the COF7 group(P<0.05),and the CD4+/CD8+T ratio also significantly increased progressively in the two groups(P<0.05).The results of RT-qPCR showed that compared with the corresponding control group,the mRNA expression levels of NF-κB,NLRP3,Caspase 1,and IL-1β in adipose tissue of mice in the COF3 group and the COF7 group were significantly increased(P<0.05,P<0.01).The mRNA expression levels of mice in each exposure group gradually increased over time.The Western blot results showed that compared with the corresponding control group,the protein expressions of NLRP3 and Caspase 1 in the COF3 group were significantly increased(P<0.01),and the expression of IL-1β protein also increased but without statistical significance.The protein expressions of NLRP3,Caspase 1,and IL-1β in the COF7 group were significantly higher than those in the CON7 group(P<0.05,P<0.01). [Conclusion]Acute exposure to cooking oil fumes can induce significant inflammatory response in adipose tissue,and the effect gradually increases with the extension of exposure time.The mechanism of action may be related to the activation of NLRP3 inflammasome signaling pathway.

王鸽;吴彪;郭建树;范冬霞;于路;张炽航;糜岚;王丽娜;赵金镯

复旦大学 公共卫生学院环境卫生学教研室,上海 200030复旦大学 光科学与工程系,上海 200030复旦大学 环境科学与工程系,上海 200030复旦大学 公共卫生学院环境卫生学教研室,上海 200030||复旦大学 IRDR极端天气气候与健康风险互联和治理国际卓越中心,上海 200030

预防医学

烹调油烟脂肪组织炎症反应NLRP3Caspase1IL-1β信号通路

cooking fumeadipose tissueinflammatory responseNLRP3Caspase 1IL-1βsignaling pathway

《环境与职业医学》 2024 (004)

349-355 / 7

10.11836/JEOM23339

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