藻蓝蛋白通过调节NF-κB通路影响游离脂肪酸诱导的非酒精性脂肪肝细胞模型  

Phycocyanin Regulates NF-κB Signaling Pathway to Affect FFA-Induced Non-Alcoholic Fatty Liver Cell Model

作　　者：孙孟筱 郝建华[2] 王伟[2] 孙晶晶[2] 江承程 张海峰 SUN Mengxiao;HAO Jianhua;WANG Wei;SUN Jingjing;JIANG Chengcheng;ZHANG Haifeng(School of Basic Medical Science,Inner Mongolia Medical University,Hohhot 010110,China;Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao 266071,China)

机构地区：[1]内蒙古医科大学基础医学院,内蒙古呼和浩特010110 [2]中国水产科学研究院黄海水产研究所,山东青岛266071

出　　处：《青岛农业大学学报(自然科学版)》2025年第1期50-57,64,共9页Journal of Qingdao Agricultural University(Natural Science)

基　　金：中央级公益性科研院所基本科研业务费专项(2023TD71);内蒙古医科大学附属医院重点实验室开放基金(2022NYFYSYS012);内蒙古卫生委员会医疗卫生技术项目(202201236);内蒙古自治区直属大学基础研究基金(YKD2023MS045)。

摘　　要：旨在探索藻蓝蛋白(phycocyanin,PC)对游离脂肪酸(free fatty acid,FFA)诱导的非酒精性脂肪肝(non-alcoholic fatty liver disease,NAFLD)细胞模型的保护作用及其可能机制。采用FFA溶液处理人肝癌细胞株HepG2(human hepatocellular carcinoma cell),建立NAFLD细胞模型,用不同浓度的PC对HepG2细胞作用24 h,进行油红O染色并检测甘油三酯(triglyceride,TG)、总胆固醇(total cholesterol,TC)、高密度脂蛋白胆固醇(high-density lipoprotein cholesterol,HDL-C)、低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-C)、丙氨酸氨基转移酶(alanine aminotransferase,ALT)和天门冬氨酸氨基转移酶(aspartate aminotransferase,AST)的水平,检测核因子κB(nuclear factor kappa-B,NF-κB)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)和白细胞介素1β(interleukin-1β,IL-1β)的mRNA表达水平,检测TNF-α、NF-κB亚基p65、核因子κB抑制因子激酶(inhibitor of kappa B kinase,IKK)、核因子κB抑制蛋白(inhibitor of NF-κB,IκB)等的总蛋白及相关磷酸化蛋白表达水平。油红O染色结果显示,PC处理FFA诱导的HepG2细胞后,与模型组相比,试验组细胞内脂质堆积情况明显改善,TC、TG、LDL-C、ALT、AST水平下降,HDL-C水平上升(P<0.05)。定量逆转录聚合酶链式反应(reverse transcription quantitative polymerase chain reaction,RT-qPCR)结果显示,与模型组相比,试验组NF-κB、TNF-α、IL-1β、IL-6的mRNA表达水平下降(P<0.05)。而免疫印迹试验(immunoblotting)结果则显示试验组TNF-α、p-NF-κB p65/NF-κB p65、p-IKKβ/IKKβ、p-IκBα/IκBα蛋白表达水平下降(P<0.05)。表明PC对NAFLD细胞模型有一定的保护作用,可能机制是PC降低NAFLD细胞模型中TNF-α、IL-6和IL-1β等促炎性细胞因子水平,抑制IKK、IκB和NF-κB p65磷酸化。这提示PC可能通过抑制NF-κB信号通路抑制炎症反应,减轻NAFLD影响。The aim of this study was to explore the protective effect of phycocyanin(PC)on free fatty acid(FFA)-induced non-alcoholic fatty liver disease(NAFLD)cell model and its possible mechanism.Human hepatocellular carcinoma cell line(HepG2)was treated with a FFA solution to establish a NAFLD cell model.HepG2 cells were treated with various concentrations of PC for 24 hours.Levels of triglyceride(TG),total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C),alanine aminotransferase(ALT),and aspartate aminotransferase(AST)were assessed using Oil Red O staining.The mRNA expression levels of nuclear factor kappa-B(NF-κB),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-1β(IL-1β)were detected.Expression levels of total proteins such as TNF-α,NF-κB p65,inhibitor of kappa B kinase(IKK),inhibitor of NF-κB(IκB),and related phosphorylated proteins were determined.Results of Oil Red O staining showed that after PC treatment of FFA-induced HepG2 cells,the lipid accumulation in the experimental groups significantly improved compared with the model groups.Levels of TC,TG,LDL-C,ALT and AST decreased,while the HDL-C level increased(P<0.05).Results of reverse transcription quantitative polymerase chain reaction(RT-qPCR)showed that mRNA expression levels of NF-κB,TNF-α,IL-1β,and IL-6 in experimental groups decreased compared with model groups(P<0.05).Results of immunoblotting test indicated a decrease in expression levels of TNF-α,p-NF-κB p65/NF-κB p65,p-IKKβ/IKKβ,and p-IκBα/IκBαproteins in experimental groups(P<0.05).All results showed that PC had a protective effect on the NAFLD cell model.The possible mechanism is that PC can reduce levels of proinflammatory cytokine such as TNF-α,IL-6 and IL-1βin the NAFLD cell model,and inhibit phosphorylation of IKK,IκB and NF-κB p65.This suggests that PC may inhibit inflammatory reaction by regulating NF-κB signaling pathway to mitigate the impact of NAFLD.

关 键 词：非酒精性脂肪性肝病 藻蓝蛋白 NF-ΚB 

分 类 号：R471.5[医药卫生—护理学]