畜牧兽医学报2025，Vol.56Issue(4)：1919-1933,15.DOI:10.11843/j.issn.0366-6964.2025.04.039
大麻二酚通过BRD4/AMPK/mTOR信号通路拮抗双酚A诱导的猪肠上皮细胞凋亡和自噬
Cannabidiol Antagonizes BPA-induced Apoptosis and Autophagy in Porcine Intestinal Epithelial Cells through the BRD4/AMPK/mTOR Signaling Pathway
摘要
Abstract
This study aimed to explore the mechanism of cannabidiol(CBD)in alleviating the ap-optosis and autophagy of pig intestinal epithelial cells(IPEC-J2)induced by bisphenol A(BPA)through BRD4/AMPK/mTOR signaling pathway.In this study,the cell viability was measured by CCK-8 method,and the half inhibitory concentration of BPA on IPEC-J2 cells and the optimal dose of CBD antagonist for BPA were screened.Using the group comparison method,The exper-iment was divided into blank control group,BPA group(150 μmol·L-1 BPA),BPA+CBD group(150 μmol·L-1 BPA+10 μmol·L-1 CBD),BPA+CBD+CQ group(150 μmol·L-1 BPA+10 μmol·L-1 CBD+20 μmol·L-1 CQ)and CBD group(10 μmol·L-1 CBD).The apop-tosis rate of IPEC-J2 cells was detected by AO/EB staining and flow cytometry.The ROS levels in cells were detected by DCFH-DA method.Oxidative stress was detected by oxidative stress kit.The expression of LC3-Ⅱ protein was detected by immunofluorescence technique.Quantita-tive Real-time PCR(qRT-PCR)and Western blot were used to detect the expression of genes re-lated to apoptosis,autophagy,intestinal tight-junction protein and BRD4/AMPK/mTOR signa-ling pathway.The results showed that apoptosis and autophagy levels of IPEC-J2 cells were in-creased after 150 μmol·L-1 BPA treatment,and decreased after 10 μmol·L-1 CBD treatment.CBD could significantly down-regulate the increase of ROS and MDA levels caused by BPA(P＜0.05),and up-regulate the decrease of GSH-Px activity(P＜0.05);CBD significantly down-reg-ulated the increase of expression levels of apoptosis related genes(Bax and caspase-3),autophagy related genes(P62,LC3-Ⅱ/LC3-Ⅰ and ATG5)and BRD4/AMPK/mTOR path-related genes(AMPK)induced by BPA(P＜0.05).The expression levels of LAMP1,Bcl-2,BRD4,mTOR,ZO-1 and Claudin-1 were increased(P＜0.05);Immunofluorescence results showed that CBD could significantly reduce the expression and distribution of LC3-Ⅱ in IPEC-J2 cells induced by BPA(P＜0.05).In conclusion,CBD could antagonize BPA-induced apoptosis and autophagy of IPEC-J2 cells through BRD4/AMPK/mTOR signaling pathway.关键词
双酚A/大麻二酚/猪肠上皮细胞/细胞凋亡/细胞自噬/BRD4/AMPK/mTORKey words
bisphenol A/cannabidiol/porcine intestinal epithelial cells/apoptosis/autophagy/BRD4/AMPK/mTOR分类
畜牧业引用本文复制引用
侯宛辰,徐童..大麻二酚通过BRD4/AMPK/mTOR信号通路拮抗双酚A诱导的猪肠上皮细胞凋亡和自噬[J].畜牧兽医学报,2025,56(4):1919-1933,15.基金项目
东北农业大学"青年才俊"项目 （）
2023年国家级大学生创新创业训练计划项目(SIPT项目编号:202310224097) （SIPT项目编号:202310224097）
