实用医学杂志 ›› 2021, Vol. 37 ›› Issue (22): 2856-2861.doi: 10.3969/j.issn.1006⁃5725.2021.22.006

• 基础研究 • 上一篇    下一篇

利拉鲁肽联合锌alpha2糖蛋白对棕榈酸诱导的HepG2细胞凋亡、脂质代谢和炎症因子表达的影响

范明娟 聂玉强 詹琪 李洁   

  1. 广州市第一人民医院老年病科(广州 510180)

  • 出版日期:2021-11-25 发布日期:2021-11-25
  • 基金资助:
    广东省自然科学基金项目(编号:2016A030310110)

Effects of liraglutide combined with ZAG on palmitic acid ⁃ induced apoptosis,lipid metabolism and expression of inflammatory factors in HepG2 cells

FAN Mingjuan,NIE Yuqiang,ZHAN Qi,LI Jie.   

  1. Department of Geriatrics,Guangzhou Frist People′s Hospital,Guangzhou 510180,China

  • Online:2021-11-25 Published:2021-11-25

摘要:

目的 探讨利拉鲁肽联合锌 alpha2 糖蛋白(ZAG)对棕榈酸诱导的 HepG2 细胞凋亡、脂质代谢和炎症因子表达的影响。方法 实时定量 PCR(RT⁃qPCR)和蛋白质印迹法分析 ZAG 在非酒精性脂肪性肝病(NALFD)患者肝组织以及棕榈酸诱导的 HepG2 细胞中的表达水平。将 HepG2 细胞分为对照组、棕榈酸组、棕榈酸+利拉鲁肽组、棕榈酸+pcDNA⁃ZAG 组、棕榈酸+利拉鲁肽+ZAG shRNA 组、棕榈酸+ 利拉鲁肽+pcDNA⁃ZAG 组。流式细胞术分析细胞凋亡情况。试剂盒测定细胞中总胆固醇(TC)、甘油三 酯(TG)水平,同时测定细胞培养液上清中白细胞介素 6(IL⁃6)、肿瘤坏死因子 α(TNF⁃α)水平。结果 NALFD 患者肝组织中 ZAG mRNA ZAG 蛋白表达水平显著降低(P < 0.05)。棕榈酸处理显著下调 ZAG mRNA ZAG 蛋白表达,增加 TC、TG、IL⁃6 TNF⁃α 水平,增加 HepG2 细胞凋亡率(P < 0.05)。利拉鲁肽处理或过表达 pcDNA⁃ZAG 显著抑制棕榈酸诱导的 HepG2 细胞凋亡,并降低 TC、TG、IL⁃6 TNF⁃α 水平 P < 0.05)。沉默 ZAG 显著减弱利拉鲁肽对棕榈酸诱导的 HepG2 细胞凋亡、TC、TG、IL⁃6 TNF⁃α 水平的 影响(P < 0.05)。过表达 ZAG 显著增强利拉鲁肽对棕榈酸诱导的 HepG2 细胞凋亡、TC、TG、IL⁃6 TNF⁃α 水平的影响(P < 0.05)。结论 利拉鲁肽联合 ZAG 可抑制棕榈酸诱导的 HepG2 细胞凋亡、脂质沉积和炎 症反应。

关键词:

利拉鲁肽, alpha2糖蛋白, 棕榈酸, HepG2细胞, 凋亡, 炎症, 甘油三酯

Abstract:

Objective To investigate the effect of liraglutide combined with zinc alpha2 glycoprotein (ZAG)on palmitic acid⁃induced HepG2 cell apoptosis,lipid metabolism and the expression of inflammatory factor. Methods Real⁃time quantitative PCR(RT⁃qPCR)and western blotting were used to analyze the expression levels of ZAG in liver tissues of patients with non⁃alcoholic fatty liver disease(NALFD)and palmitic acid⁃induced HepG2 cells. HepG2 cells were divided into control group,palmitic acid group,palmitic acid + liraglutide group,palmitic acid + pcDNA⁃ZAG group,palmitic acid + liraglutide + ZAG shRNA group,palmitic acid + liraglutide + pcDNA⁃ ZAG group. Cell apoptosis was detected by flow cytometry. Kit was applied to determine the levels of total cholesterol (TC)and triglycerides(TG)in the cells,as well as the levels of interleukin 6(IL⁃6)and tumor necrosis factor⁃α (TNF⁃α)in the supernatant of the cell culture medium. Results The expression of ZAG mRNA and ZAG protein in liver tissues of NALFD patients was significantly reduced(P < 0.05). Treatment with Palmitic acid significantly down⁃regulated the expression of ZAG mRNA and ZAG protein,increased the levels of TC,TG,IL⁃6 and TNF⁃α and increased the apoptosis rate of HepG2 cells(P < 0.05). Treatment with liraglutide or over⁃expression of pcDNA⁃ ZAG significantly inhibited palmitic acid⁃induced apoptosis of HepG2 cells,and reduced the levels of TC,TG,IL⁃6 and TNF ⁃α(P < 0.05). Silencing ZAG significantly weakened the effect of liraglutide on palmitic acid ⁃induced apoptosis,TC,TG,IL ⁃6 and TNF ⁃ α levels in HepG2 cells(P < 0.05). Over ⁃ expression of ZAG significantly enhanced the effect of liraglutide on palmitic acid⁃induced apoptosis,TC,TG,IL⁃6 and TNF⁃α levels in HepG2 cells(P < 0.05). Conclusion Liraglutide combined with ZAG can inhibit palmitic acid⁃induced apoptosis,lipid deposition and inflammatory reaction in HepG2 cells. 

Key words:

liraglutide, zinc alpha2 glycoprotein, palmitic acid, HepG2 cells, apoptosis, inflamMolation, triglycerides