大麻二酚对脂多糖诱导的小鼠乳腺上皮细胞炎症反应和凋亡的抑制作用

doi:10.16431/j.cnki.1671-7236.2025.01.006

摘要/Abstract

摘要： 【目的】研究大麻二酚(CBD)对脂多糖(LPS)诱导的小鼠乳腺上皮细胞(HC11)炎症反应及细胞凋亡的影响。【方法】试验将HC11细胞分为对照组、LPS组和不同浓度CBD＋LPS共处理组(CBD浓度分别为2、4、6 μmol/L)。对照组细胞仅加入培养液,LPS组加入LPS处理,CBD＋LPS共处理组用不同浓度CBD预处理1 h后加入LPS,24 h后收集各组细胞样品。利用ELISA法检测细胞中炎症因子含量;使用实时荧光定量PCR、Western blotting分别检测细胞炎症因子及核因子κB(NF-κB)、丝裂原活化蛋白激酶(MAPK)信号通路相关基因mRNA和蛋白的表达水平;通过AnnexinV-FITC/碘化丙啶(PI)和JC-1染色法分别检测细胞凋亡率和线粒体膜电位水平。【结果】与对照组相比,LPS组细胞中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、IL-6含量及mRNA表达水平均极显著或显著升高(P＜0.01;P＜0.05),NF-κB p65、核因子-κB抑制蛋白α(IκBα)、细胞外信号调节激酶(ERK)、丝裂原活化蛋白激酶(p38)、应激活化蛋白激酶(JNK)的mRNA表达量和蛋白磷酸化水平极显著或显著升高(P＜0.01;P＜0.05)。与LPS组相比,不同浓度CBD与LPS共处理后细胞中TNF-α、IL-1β、IL-6含量及mRNA表达量均极显著或显著降低(P＜0.01;P＜0.05),NF-κB p65、IκBα、ERK、p38、JNK的mRNA表达量和蛋白磷酸化水平均极显著或显著降低(P＜0.01;P＜0.05)。细胞凋亡率和线粒体膜电位检测结果显示,与对照组相比,LPS组HC11细胞凋亡率显著升高(P＜0.05),线粒体膜电位显著降低(P＜0.05)。与LPS组相比,4 μmol/L CBD和LPS共处理后,HC11细胞凋亡率显著降低(P＜0.05),线粒体膜电位显著升高(P＜0.05)。【结论】在LPS诱导的小鼠HC11细胞炎症及凋亡模型中,适宜浓度CBD能抑制NF-κB和MAPK信号通路的激活,从而抑制炎症因子的释放来缓解炎症反应;同时能降低LPS诱导的HC11细胞凋亡率、上调细胞线粒体膜电位,从而抑制细胞凋亡发生。

关键词: 大麻二酚; 脂多糖; 小鼠乳腺上皮细胞; 炎症反应; 细胞凋亡

Abstract: 【Objective】 The aim of this study was to investigate the effects of cannabidiol (CBD) on lipopolysaccharide (LPS)-induced inflammatory response and apoptosis of mouse mammary epithelial cells (HC11). 【Method】 HC11 cells were divided into control group,LPS group and different concentrations of CBD＋LPS groups (CBD concentration was 2,4 and 6 μmol/L,respectively).The cells in control group were only treated with culture medium,and the cells in LPS group were treated with LPS.CBD＋LPS groups were pretreated with different concentrations of CBD for 1 h and then LPS was added.Cell samples were collected after 24 h of culture.The contents of inflammatory factors in the cells were detected by ELISA.Real-time quantitative PCR and Western blotting were used to detect the mRNA and protein expression levels of inflammatory cytokines,and genes related to nuclear factor κB (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways,respectively.Apoptosis rate and mitochondrial membrane potential were detected by AnnexinV-FITC/PI and JC-1 staining,respectively. 【Result】 Compared with control group,the contents and mRNA expression levels of tumor necrosis factor-α (TNF-α),interleukin-1β (IL-1β) and IL-6 in LPS group were extremely significantly or significantly increased (P＜0.01 or P＜0.05),the mRNA expression and protein phosphorylation levels of NF-κB p65,nuclear factor-κB inhibitory protein α (IκBα),extracellular signal-regulated kinase (ERK),mitogen-activated protein kinase (p38),and stress-activated protein kinase (JNK) were extremely significantly or significantly increased (P＜0.01 or P＜0.05).Compared with LPS group,the contents and mRNA expression of TNF-α,IL-1β and IL-6 in cells treated with different concentrations of CBD and LPS were extremely significantly or significantly decreased (P＜0.01 or P＜0.05). The mRNA expression and protein phosphorylation levels of NF-κB p65,IκBα,ERK,p38 and JNK were extremely significantly or significantly decreased (P＜0.01 or P＜0.05).The results of apoptosis rate and mitochondrial membrane potential detection showed that,compared with control group,the apoptosis rate of HC11 cells in LPS group was significantly increased (P＜0.05),and the mitochondrial membrane potential was significantly decreased (P＜0.05). Compared with LPS group,the apoptosis rate of HC11 cells was significantly decreased after 4 μmol/L CBD and LPS co-treatment (P＜0.05),and the mitochondrial membrane potential was significantly increased (P＜0.05). 【Conclusion】 In the LPS-induced inflammation and apoptosis model of HC11 cells,the appropriate concentration of CBD could inhibit the activation of NF-κB and MAPK signaling pathways,thereby inhibiting the release of inflammatory factors to alleviate inflammation.Furthermore,it could reduce the apoptosis rate and up-regulate the mitochondrial membrane potential of HC11 cells induced by LPS,thus inhibiting the cell apoptosis.

Key words: cannabidiol; lipopolysaccharide; mouse mammary epithelial cells; inflammatory response; apoptosis

中图分类号:

S853.74

常欣欣, 黄鑫, 范港, 杨佳欣, 张伟伟, 杨清竹, 田哲. 大麻二酚对脂多糖诱导的小鼠乳腺上皮细胞炎症反应和凋亡的抑制作用[J]. 中国畜牧兽医, 2025, 52(1): 60-69.

CHANG Xinxin, HUANG Xin, FAN Gang, YANG Jiaxin, ZHANG Weiwei, YANG Qingzhu, TIAN Zhe. Inhibitory Effect of Cannabidiol on LPS-induced Inflammatory Response and Apoptosis of Mouse Mammary Epithelial Cells[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(1): 60-69.

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