盐酸小檗碱缓解高胆固醇所致肝脏损伤的机制体外及动物实验研究

doi:10.16150/j.1671-2870.2018.03.016

摘要/Abstract

摘要： 目的：采用动物实验,探讨盐酸小檗碱（berberine,BBR）对高胆固醇所致肝脏损伤的保护作用。方法：将8周龄雄性C57BL/6J小鼠随机分为3组（每组10只）,分别给予普通饲料组、高胆固醇饲料组（高胆固醇组）和高胆固醇饲料加BBR治疗组（以下简称BBR组）,喂养8周后采集其肝脏和血清。采用反转录聚合酶链反应（reverse transcription polymerase chain reaction, RT-PCR）法测量小鼠肝脏相关基因mRNA的表达,并测定肝脏组织丙二醛（malondialdehyde,MDA）的含量。采用HepG₂细胞系,研究BBR对高胆固醇处理下细胞内质网应激的影响。结果：与普通饲料组比较,高胆固醇组小鼠的肝脏组织中CHOP、GRP78、ATF4等内质网应激相关基因mRNA表达量增加了1倍以上（P<0.05）,Trp53、Nqo1等氧化应激相关基因的mRNA表达量分别增加了8倍、5倍（P<0.05）,肿瘤坏死因子α、白细胞介素1β等炎症因子相关基因的mRNA表达量分别增加了15倍和0.5倍（P<0.05）,而BBR组的上述基因表达与普通饲料组水平接近（P<0.05）。高胆固醇组小鼠肝脏的MDA含量增加了1倍以上（P<0.05）,而BBR可下调高胆固醇导致的肝脏MDA含量增高（P<0.05）。HepG₂细胞体外实验显示,BBR可降低高胆固醇导致的内质网应激关键蛋白GRP78表达上调（P<0.05）和二氢乙啶（dihydroethidium,DHE）荧光量上调。结论：BBR可缓解高胆固醇导致的肝脏细胞内质网应激、氧化应激及炎症反应,具有一定的肝脏保护作用。

关键词: 盐酸小檗碱, 胆固醇, 内质网应激, 氧化应激, 炎症

Abstract: Objective: To investigate the protective effect of berberine (BBR) on liver damage under high-cholesterol diet challenge. Methods: Eight-week old male C57BL/6J mice were randomly divided into normal diet group, high cholesterol diet (1.25% cholesterol+0.5% cholic acid) group and high cholesterol diet+BBR (100 mg/kg per day by gavage) group, with 10 mice in each group. Liver and serum plasma samples were collected after 8-week feeding. Hepatic mRNA expressions of genes involved in endoplasmic reticulum stress (ER stress), oxidative stress and inflammatory cytokines were determined by real-time quantitative PCR, and content of malondialdehyde (MDA) in liver tissues was measured. HepG₂ cell line was used to investigate the effect of BBR on ER stress under high cholesterol challenge. Results: Compared with normal diet group, hepatic mRNA expressions of genes in ER stress, oxidative stress and inflammatory cytokines increased significantly in high cholesterol diet group (P<0.05), and were reduced closely to levels of normal diet group by BBR treatment (P<0.05). BBR also down-regulated the increased hepatic MDA content induced by high cholesterol diet (P<0.05). Furthermore, BBR decreased the expression of GRP78, a key protein marker concerning ER stress (P<0.05), as well as the fluorescence intensity of DHE probe after high cholesterol treatment in HepG₂ cells. Conclusions: BBR could effectively ameliorate hepatic ER stress, oxidative stress and inflammatory response under high cholesterol challenge, which might protect liver from damage when overloaded with cholesterol.

Key words: Berberine, Cholesterol, ER stress, Oxidative stress, Inflammation

中图分类号:

R575

邵文涛, 孙海东, 王起晗, 刘倩, 蒋兆彦, 顾爱华. 盐酸小檗碱缓解高胆固醇所致肝脏损伤的机制体外及动物实验研究[J]. 诊断学理论与实践, 2018, 17(03): 311-317.

SHAO Wentao, SUN Haidong, WANG Qihan, LIU Qian, JIANG Zhaoyan, GU Aihua. Animal and in vitro study on mechanism of berberine alleviating high cholesterol induced liver injury[J]. Journal of Diagnostics Concepts & Practice, 2018, 17(03): 311-317.

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