青钱柳通过增强ABCG5/8和SREBP2调节MASLD小鼠的胆固醇代谢

doi:10.5246/jcps.2025.04.023

中国药学(英文版) ›› 2025, Vol. 34 ›› Issue (4): 305-320.DOI: 10.5246/jcps.2025.04.023

• 【研究论文】 • 下一篇

青钱柳通过增强ABCG5/8和SREBP2调节MASLD小鼠的胆固醇代谢

包小艾¹^,²^,^#, 杨娇¹^,²^,^#, 刘晓艳², 孙义政², 许海龙¹^,², 韩蓉²^,³, 朱海涛⁴, 邓改改¹^,^*(), 张友波²^,⁵^,^*()

^1. 三峡大学健康医学院国家中医药管理局中药药理三级实验室, 湖北宜昌 443002
^2. 北京大学药学院天然药物学系, 天然药物及仿生药物全国重点实验室, 北京 100191
^3. 兰州理工大学生命科学与工程学院, 甘肃兰州 730050
^4. 山东省德州市临邑县人民医院影像科, 山东德州 251500
^5. 河南中医药大学药食两用中药技术河南工程研究中心, 河南郑州 450046

收稿日期:2024-11-06 修回日期:2025-01-23 接受日期:2025-03-06 出版日期:2025-05-02 发布日期:2025-05-02
通讯作者: 邓改改, 张友波

Cyclocarya paliurus modulates cholesterol metabolism in MASLD mice via upregulation of ABCG5/8 and SREBP2

Xiaoai Bao¹^,²^,^#, Jiao Yan¹^,²^,^#, Xiaoyan Liu², Yizheng Sun², Hailong Xu¹^,², Rong Han²^,³, Haitao Zhu⁴, Gaigai Deng¹^,^*(), Youbo Zhang²^,⁵^,^*()

¹ College of Medicine and Health Sciences, State Administration of Traditional Chinese Medicine, China Three Gorges University, Yichang 443002, Hubei, China
² State Key Laboratory of Natural and Biomimetic Drugs, Department of Natural Medicines, School of Pharmaceutical Sciences, Peking University, Beijing 100191, China
³ School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, Gansu, China
⁴ Department of Imaging, Linyi County People’s Hospital, Dezhou 251500, Shandong, China
⁵ Henan Engineering Research Center of Medicinal and Edible Chinese Medicine Technology, Henan University of Chinese Medicine, Zhengzhou 450046, Henan, China

Received:2024-11-06 Revised:2025-01-23 Accepted:2025-03-06 Online:2025-05-02 Published:2025-05-02
Contact: Gaigai Deng, Youbo Zhang
About author:
^# Xiaoai Bao and Jiao Yang contributed equally to this work.
Supported by:
National Key Research and Development Program of China (Grant No. 2022YFC3501700) and the Beijing Municipal Natural Science Foundation (Grant No. 7144219).

摘要/Abstract

摘要：

代谢功能障碍相关性脂肪性肝病(MASLD)已成为世界范围内慢性肝病的主要病因, 且发生率逐年增加。如果不进行治疗, MASLD可能会发展为代谢功能障碍在相关的脂肪性肝炎(MASH), 这是一种更严重的疾病, 可不可逆地进展为肝纤维化、肝硬化, 甚至肝细胞癌(HCC)。最近的研究表明胆固醇代谢失调与MASLD的发病机制和严重程度之间存在密切联系。这强调了全面探索MASLD肝脏胆固醇代谢调控机制的迫切需要, 因为这些见解可以揭示新的治疗靶点, 并为早期诊断和有效预防策略铺平道路。青钱柳(Cyclocarya paliurus (Batal.) Iljinskaja)是一种药用和食用植物, 具有多种药理学特性, 包括降血糖、调节血脂和保肝作用。本研究旨在探讨青钱柳提取物(CCE)对甲硫氨酸胆碱缺乏(MCD)诱导的MASLD小鼠模型的降血脂和保肝作用。以辛伐他汀为阳性对照药, 同时给予不同剂量的CCE以评估其治疗潜力。对照组和模型组同时给予0.5%羧甲基纤维素钠(CMC-Na)灌胃, 每日1次, 连续6周。在给药期结束后, 采集血液和肝脏样本进行生化分析、组织病理学评估和基因表达谱分析。结果表明, 青钱柳提取物(CCE)可显著降低大鼠血清中谷草转氨酶(AST)、谷丙转氨酶(ALT)含量, 增加胆碱酯酶(CHE)、高密度脂蛋白胆固醇(HDL-C)的活性。在肝组织中, CCE可显著降低总胆固醇(Total Cholesterol, TC)和甘油三酯(TG)的水平, 同时增加肝HDL-C的含量。组织学分析显示, CCE处理小鼠的病理性肝损伤显著减轻。分子研究进一步表明, CCE下调了参与胆固醇合成的关键基因和蛋白质的表达, 包括SREBP2、LDLR和HMGCR。同时, 它上调了与胆固醇转运相关的基因和蛋白质的表达, 例如ABCG5和ABCG8。此外, CCE通过提高促炎细胞因子(包括TNF-α和IL-6)的表达水平以及调节氧化应激标志物(如NRF2、KEAP1和NQO1)来缓解炎症。蛋白质表达分析显示IL-6和IL-1β水平降低, 进一步证实了其抗炎作用。综上所述, 青钱柳对MCD诱导的MASLD小鼠表现出明显的肝保护作用。这些保护机制可能与胆固醇转运蛋白ABCG5/8的上调和固醇调节元件结合蛋白2(SREBP2)的调节密切相关。本研究强调了青钱柳作为MASLD的干预措施的治疗潜力, 并强调了其在调节胆固醇代谢和减轻炎症和氧化应激方面的作用。

关键词: 青钱柳, MASLD, 胆固醇, ABCG5/8, SREBP2

Abstract:

Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD) has emerged as a predominant cause of chronic liver disease globally, with its prevalence rising steadily each year. If left untreated, MASLD may progress to metabolic dysfunction in associated steatohepatitis (MASH), a more severe condition that can irreversibly advance to liver fibrosis, cirrhosis, and even hepatocyte carcinoma (HCC). Recent studies have illuminated a pivotal link between dysregulated cholesterol metabolism and the pathogenesis and severity of MASLD. This underscores the critical need for a comprehensive exploration of the regulatory mechanisms underlying hepatic cholesterol metabolism in MASLD, as such insights could unveil new therapeutic targets and pave the way for early diagnosis and effective prevention strategies. Cyclocarya paliurus (Batal.) Iljinskaja, a plant known for both medicinal and dietary applications, has demonstrated diverse pharmacological properties, including hypoglycemic, lipid-regulating, and hepatoprotective effects. This study aimed to investigate the hypolipidemic and hepatoprotective activities of Cyclocarya paliurus extract (CCE) in a murine model of MASLD induced by a methionine-choline-deficient (MCD) diet. Simvastatin was employed as a positive control drug, while various doses of CCE were administered to assess its therapeutic potential. Meanwhile, the control and model groups received 0.5% sodium carboxymethyl cellulose (CMC-Na) once daily for 6 weeks. At the end of the treatment period, blood and liver samples were collected for biochemical analysis, histopathological assessment, and gene expression profiling. The findings revealed that CCE significantly reduced serum levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) while enhancing the activities of cholinesterase (CHE) and high-density lipoprotein cholesterol (HDL-C). In liver tissues, CCE markedly decreased the levels of total cholesterol (TC) and triglycerides (TG), while simultaneously increasing hepatic HDL-C content. Histological analyses showed notable alleviation of pathological liver damage in CCE-treated mice. Molecular studies further demonstrated that CCE downregulated the expression of key genes and proteins involved in cholesterol synthesis, including SREBP2, LDLR, and HMGCR. Concurrently, it upregulated the expression of genes and proteins related to cholesterol transport, such as ABCG5 and ABCG8. Additionally, CCE mitigated inflammation by improving the expression levels of pro-inflammatory cytokines, including TNF-α and IL-6, and modulated oxidative stress markers, such as NRF2, KEAP1, and NQO1. Protein expression analyses revealed reduced levels of IL-6 and IL-1β, further corroborating its anti-inflammatory effects. In summary, C. paliurus exhibited potent hepatoprotective effects in MCD-induced MASLD mice. These protective mechanisms were closely linked to the upregulation of cholesterol transporters ABCG5/8 and the modulation of sterol regulatory element-binding protein 2 (SREBP2). This study highlighted the therapeutic potential of C. paliurus as a promising intervention for MASLD and underscored its role in regulating cholesterol metabolism and mitigating inflammation and oxidative stress.

Key words: Cyclocarya paliurus, MASLD, Cholesterol, ABCG5/8, SREBP2

Supporting:

包小艾, 杨娇, 刘晓艳, 孙义政, 许海龙, 韩蓉, 朱海涛, 邓改改, 张友波. 青钱柳通过增强ABCG5/8和SREBP2调节MASLD小鼠的胆固醇代谢[J]. 中国药学(英文版), 2025, 34(4): 305-320.

Xiaoai Bao, Jiao Yan, Xiaoyan Liu, Yizheng Sun, Hailong Xu, Rong Han, Haitao Zhu, Gaigai Deng, Youbo Zhang. Cyclocarya paliurus modulates cholesterol metabolism in MASLD mice via upregulation of ABCG5/8 and SREBP2[J]. Journal of Chinese Pharmaceutical Sciences, 2025, 34(4): 305-320.

图/表 12

Table 1. Sequences of the primers.

Figure 1. Effects of C. paliurus on liver index and MAS score of MASLD mice. (A) Effects of C. paliurus on liver index of MASLD mice; (B) Effects of C. paliurus on MAS score of MASLD mice. Results are presented as mean ± SEM (n = 10). *P < 0.05, **P < 0.01 compared with the MCS group and #P < 0.05, ##P < 0.01 compared with the MCD group.

Figure 2. Effects of C. paliurus on serum biochemical parameters of MASLD mice. Columns showed ALT level (A), AST level (B), CHE level (C), and HDL-C level (D) in MASLD mice serum. Results are presented as mean ± SEM (n = 10). ****P < 0.001 or **P < 0.01 compared with the MCS group; #P < 0.05 or ##P < 0.01 compared with the MCD group.

Table 2. Effects of C. paliurus on liver related indexes of MASLD mice (mean ± SEM).

Figure 3. Effects of C. paliurus on liver biochemical parameters of MASLD mice. Columns showed TC level (A), TG level (B), LDL-C level (C), and HDL-C level (D) in MASLD mice liver. Results are presented as mean ± SEM (n = 10). *P < 0.05 or **P < 0.01 compared with the MCS group; #P < 0.05 or ##P < 0.01 compared with the MCD group.

Table 3. Effects of C. paliurus on liver related indexes of MASLD mice (mean ± SEM).

Figure 4. Histological staining of MASLD mice (40×). (A) Control group MCS; (B) Model group MCD; (C) SV group; (D) CCE-L group; (E) CCE-M group; (F) CCE-H group.

Figure 5. Effects of C. paliurus on the mRNA expression of ABCG5/8 in MASLD mice. (A) ABCG5 level in MASLD mice; (B) ABCG8 level in MASLD mice. Results are presented as mean ± SEM (n = 10). *P < 0.05, **P < 0.01 compared with the MCS group, #P < 0.05, ##P < 0.01, ###P < 0.001 compared with the MCD group.

Figure 6. Effects of C. paliurus on the mRNA expression of SREBP2, HMGCR, LDLR, NRF2, KEAP1, and NQO1 in MASLD mice. Results are presented as mean ± SEM (n = 10). *P < 0.05, **P < 0.01 compared with the MCS group, #P < 0.05, ##P < 0.01, ###P < 0.001 compared with the MCD group.

Figure 7. Effects of C. paliurus on the mRNA expression of IL-6 and TNF-α in MASLD mice. (A) IL-6 level in MASLD mice; (B) TNF-α level in MASLD mice. Results are presented as mean ± SEM (n = 10). *P < 0.05, **P < 0.01 compared with the MCS group, #P < 0.05, ##P < 0.01, ###P < 0.001 compared with the MCD group.

Figure 8. Effects of C. paliurus on the protein expression of ABCG5/8 in MASLD mice. (A) Hepatic protein expression of ABCG5/8, β-actin served as loading control; (B) Densitometric analyses of ABCG5; (C) Densitometric analyses of ABCG8. Results are presented as mean ± SEM (n = 3). **P < 0.01 compared with the MCS group, #P < 0.05 or ##P < 0.01 compared with the MCD group.

Figure 9. Effects of C. paliurus on the protein expression of IL-1β and IL-6 in MASLD mice. (A) Hepatic protein expression of IL-6 and IL-1β, β-actin served as loading control; (B) Densitometric analyses of IL-1β; (C) Densitometric analyses of IL-6. Results are presented as mean ± SEM (n = 3). **P < 0.01 compared with the MCS group, #P < 0.05 or ##P < 0.01 compared with the MCD group.

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青钱柳通过增强ABCG5/8和SREBP2调节MASLD小鼠的胆固醇代谢

Cyclocarya paliurus modulates cholesterol metabolism in MASLD mice via upregulation of ABCG5/8 and SREBP2

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