β-胡萝卜素对脂多糖诱导的肠隐窝上皮细胞IEC-6自噬的作用机制  

Effect of β-Caroteneon the Autophagy Mechanism of Intestinal Recess Epithelial Cells IEC-6 Induced by Lipopolysaccharide

作　　者：许光[1] 马体栋[1] 赵凡[1] 李碧香[1] 周崇高[1] XU Guang;MA Ti-dong;ZHAO Fan(Department of Neonatal Surgery,Hunan Children's Hospital,Changsha Hunan 410007)

机构地区：[1]湖南省儿童医院新生儿外科,湖南长沙410007

出　　处：《医学临床研究》2022年第3期324-327,331,共5页Journal of Clinical Research

基　　金：湖南省卫生健康委科研课题(20200494);湖南省出生缺陷协同防治科技重大专项(2019SK1010)。

摘　　要：【目的】探讨β-胡萝卜素(βC)对脂多糖诱导的肠隐窝上皮细胞IEC-6自噬的作用机制。【方法】应用脂多糖(LPS)诱导肠上皮细胞株IEC-6构建新生儿坏死性小肠结肠炎(NEC)体外模型,使用PI3K抑制剂Voxtalisib(Vox)分析βC调控自噬的潜在机制,IEC-6细胞常规体外培养后分为Control组、LPS组、LPS+βC组和LPS+βC+Vox组。CCT-8检测细胞活性,转染mRFPGFPLC3自噬双标腺病毒检测细胞内自噬,Westernblotting法检测自噬相关蛋白和PI3K/AKT/mTOR信号通路相关蛋白表达情况。【结果】相对于Control组,LPS组细胞活性明显降低(P<0.05),LC3Ⅱ/Ⅰ明显增加(P<0.05),P62蛋白表达量明显降低(P<0.05),自噬体斑点数量明显增加(P<0.05)。相对于LPS组,LPS+βC组细胞活性明显增加(P<0.05),LC3Ⅱ/Ⅰ明显降低(P<0.05),P62、p-PI3K、p-AKT和p-mTOR的蛋白表达量明显增加(P<0.05),自噬体斑点数量明显减少(P<0.05)。相对于LPS+βC组,LPS+βC+Vox组细胞活性明显降低(P<0.05),LC3Ⅱ/Ⅰ明显增加(P<0.05),P62、p-PI3K、p-AKT和p-mTOR的蛋白表达量明显减少(P<0.05),自噬体斑点数量明显增加(P<0.05)。【结论】βC能够减轻LPS诱导的IEC-6细胞自噬,其作用机制与激活PI3K/AKT/mTOR信号通路有关。【Objective】To explore the effect ofβ-Carotene(βC)on the autophagy mechanism of intestinal recess epithelial cells IEC-6induced by lipopolysaccharide.【Methods】The intestinal epithelial cell line IEC-6 was induced by lipopolysaccharide(LPS)to construct the in vitro model of neonatal necrotizing enterocolitis(NEC).The PI3K inhibitor voxtalisib(VOX)was used to analyze the potential mechanism ofβC regulating autophagy.IEC-6 cells were divided into control group,LPS group and LPS+βC group and LPS+βC+Vox group after routine culture in vitro.CCC-8 was used to detect cell activity.mRFPGFPLC3 autophagy double labeled adenovirus was transfected to detect intracellular autophagy.The expressions of autophagy related proteins and PI3K/Akt/mTOR signal pathway related proteins were detected by Western blotting.【Rresults】Compared with the control group,the cell activity of LPS group decreased significantly(P<0.05),LC3Ⅱ/Ⅰincreased significantly(P<0.05),the expression of P62 protein decreased significantly(P<0.05),and the number of autophagy spots increased significantly(P<0.05).Compared with the control group,the cell activity in the LPS+βC group increased significantly(P<0.05),LC3Ⅱ/Ⅰdecreased significantly(P<0.05),the protein expression of P62 increased significantly(P<0.05),and the number of autophagy spots decreased significantly(P<0.05).Compared with the LPS group,the cell activity in the LPS+βC group increased significantly(P<0.05),LC3Ⅱ/Ⅰdecreased significantly(P<0.05).The protein expression of P62,p-PI3K,p-AKT and p-mTOR increased significantly(P<0.05),and the number of autophagy spots decreased significantly(P<0.05).【Conclusion】βC can reduce the autophagy of IEC-6 cells induced by LPS,and its mechanism is related to the activation of PI3K/AKT/mTOR signaling pathway.

关 键 词：Β胡萝卜素 脂多糖类 肠细胞/细胞学 自噬 

分 类 号：R285.5[医药卫生—中药学]