基于体内实验和网络药理学探究清肝二十七味丸对非酒精性脂肪性肝炎的作用及作用机制

doi:10.12092/j.issn.1009-2501.2026.05.003

中国临床药理学与治疗学 ›› 2026, Vol. 31 ›› Issue (5): 596-606.doi: 10.12092/j.issn.1009-2501.2026.05.003

基于体内实验和网络药理学探究清肝二十七味丸对非酒精性脂肪性肝炎的作用及作用机制

高晓艳¹(), 郭娟¹, 黄圣¹, 王巧云¹, 黄俣佳¹, 李金媛¹, 刘晓强²^,*()

1. 赤峰大学基础医学院内蒙古人类遗传病研究自治区高等学校重点实验室，赤峰 024000，内蒙古
2. 赤峰市医院，赤峰 024000，内蒙古

收稿日期:2025-07-01 修回日期:2026-01-06 出版日期:2026-05-26 发布日期:2026-06-02
通讯作者: 刘晓强 E-mail:gaoxiaoyan@cfxy.edu.cn;1365264109@qq.com
作者简介:高晓艳，女，博士研究生，讲师，从事中药改善非酒精性脂肪病等代谢性疾病药理作用机制研究。E-mail：gaoxiaoyan@cfxy.edu.cn
基金资助:
内蒙古自治区自然科学基金资助项目（2024QN08069）；赤峰学院2026年度博士科研创新发展基金项目（BSJJ015）；赤峰市科协自然科学科研课题（SZR2025010）；赤峰学院大学生创新创业训练计划项目（S202410138041，202510138051）；赤峰学院2024年第二批百名博士下基层服务地方项目（2024FWDF32）

Mechanism of Qinggan Ershiqiwei Wan on non-alcoholic steatohepatitis based on network pharmacology and in vivo experiments

Xiaoyan GAO¹(), Juan GUO¹, Sheng HUANG¹, Qiaoyun WANG¹, Yujia HUANG¹, Jinyuan LI¹, Xiaoqiang LIU²^,*()

1. Key Laboratory of Research on Human Genetic Diseases at Universities of Inner Mongolia Autonomous Region, Basic Medical College, Chifeng University, Chifeng 024000, Inner Mongolia, China
2. Chifeng Municipal Hospital, Inner Mongolia, Chifeng 024000, Inner Mongolia, China

Received:2025-07-01 Revised:2026-01-06 Online:2026-05-26 Published:2026-06-02
Contact: Xiaoqiang LIU E-mail:gaoxiaoyan@cfxy.edu.cn;1365264109@qq.com

摘要/Abstract

摘要：

目的: 基于体内实验和网络药理学探究清肝二十七味丸（QGW）对非酒精性脂肪性肝炎（NASH）的作用及作用机制。方法: 利用高脂饲料（HFD）诱导NASH小鼠模型，分为正常组（CON）、模型组（HFD）、清肝二十七味丸给药组（HFD+QGW, HQG）每组8只。第7周开始，HQG组给予药物连续作用至14周，并监测各组小鼠体质量相关指数，14周结束，解剖小鼠观察肝脏外观和形态，利用H&E染色、Oil red O染色方法观察小鼠肝组织的病理形态；比色法检测血清中的甘油三酯（TG）、总胆固醇（TC）、谷草转氨酶（AST）和谷丙转氨酶（ALT）含量；利用网络药理学通过数据库筛选QGW活性成分治疗NASH的关键靶点，并对关键靶点进行基因及通路富集分析；通过酶联免疫吸附试验（ELISA）定量分析白细胞介素-1β（IL-1β）、IL-6水平，Western blot及免疫组织化学实验对MAPK信号通路进行验证。结果: 动物实验结果表明，经QGW干预模型小鼠 8 周后，可显著降低小鼠血清TG、TC、ALT、AST水平（P<0.01）；H&E染色、油红O染色观察到QGW显著改善HFD诱导肝脏组织中细胞排列紊乱、脂质蓄积和炎症浸润；网络药理学结果表明，共筛选得到18个核心成分，包括stylopine、7beta-senecioyloxyoplopa-3（14）Z，8（10）-dien-2-one、（1R，3R，4R，5S，6S）-1-acetoxy-8-angeloxoyloxy-3，4-epoxy-5-hydroxybisabola-7（14），10-dien-2-one、7beta-（3-ethyl-cis-crotonoyloxy）-14-hydroxy-notonipetranone、木犀草素等；25个核心靶点包括MAKP8、MAPK14、MAPK9、IL-6等，涉及包括MAPK信号通路在内的 20条通路；ELISA实验表明，QGW显著降低小鼠血清中IL-1β、IL-6水平（P<0.01），Western blot及免疫组织化学实验结果表明，与CON组相比，HFD组p-ERK/ERK、p-JNK/JNK、p-P38/P38蛋白水平显著升高（P<0.01），与HFD组相比，HQG组p-ERK/ERK、p-JNK/JNK、p-P38/P38蛋白水平显著降低（P<0.05，P<0.01）。结论: QGW改善HFD诱导的NASH小鼠肝脏脂质蓄积及炎症浸润，其作用机制可能与抑制MAPK信号通路相关。

关键词: 清肝二十七味丸, 非酒精性脂肪肝炎, 网络药理学, MAPK信号通路

Abstract:

AIM: Through in vivo experiments and network pharmacology, the effects and mechanisms of Qinggan Ershiqiwei Wan (QGW) on non-alcoholic steatohepatitis (NASH) was explored. METHODS: A NASH mouse model was established by feeding a high-fat diet (HFD) induction. The C57BL/6J mice were divided into three groups: the normal group (CON), the model group (HFD), and the QGW administration group (HFD+QGW, HQG), with 8 mice in each group. Starting from the 7th week, the HQG group was administered the drug continuously until the 14th week, and the body weight-related indices of each group of mice were monitored. At the end of the 14th week, the mice were dissected to observe the appearance and morphology of the liver. Pathological morphology of the liver tissue was observed using H&E staining and Oil red O staining methods. The serum levels of triglycerides (TG), total cholesterol (TC), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were detected by colorimetry. Network pharmacology was used to screen key targets of QGW active components for the treatment of NASH through databases, and gene and pathway enrichment analysis was performed on the key targets. The levels of interleukin-1β (IL-1β) and IL-6 were quantitatively analyzed by enzyme-linked immunosorbent assay (ELISA), and the MAPK signaling pathway was experimentally validated by Western blot and immunohistochemistry. RESULTS: Animal experiments demonstrated that after 8 weeks of QGW intervention in model mice, serum levels of TG, TC, ALT and AST were significantly reduced (P<0.01). H&E staining and Oil Red O staining revealed that QGW significantly ameliorated the disordered cell arrangement, lipid accumulation, and inflammatory infiltration in liver tissues induced by HFD. Network pharmacology results revealed the screening of 18 core components, including stylopine, 7β-senecioyloxyoplopa-3(14)Z, 8(10)-dien-2-one, (1R,3R,4R,5S,6S)-1-acetoxy-8-angeloxoyloxy-3, 4-epoxy-5-hydroxybisabola-7(14), 10-dien-2-one, 7β-(3-ethyl-cis-crotonoyloxy)-14-hydroxy-notonipetranone, and luteolin; the 25 core targets, including MAKP8, MAPK14, MAPK9, and IL-6, were screened, involving 20 pathways including the MAPK signaling pathway. ELISA experiments demonstrated that QGW reduced the levels of IL-1β and IL-6 in the serum of mice (P<0.01). Western blot and immunohistochemistry results indicated that, compared to the CON group, the protein levels of p-ERK/ERK, p-JNK/JNK, and p-P38/P38 were significantly increased in the HFD group (P<0.01). In comparison to the HFD group, the protein levels of p-ERK/ERK, p-JNK/JNK, and p-P38/P38 were decreased in the HQG group ( P<0.05, P<0.01). CONCLUSION: QGW alleviates hepatic lipid accumulation and inflammatory infiltration in HFD-induced NASH mice, with its mechanism likely related to the inhibition of the MAPK signaling pathway.

Key words: Qinggan Ershiqiwei Wan, NASH, network pharmacology, MAPK signaling pathway

中图分类号:

高晓艳, 郭娟, 黄圣, 王巧云, 黄俣佳, 李金媛, 刘晓强. 基于体内实验和网络药理学探究清肝二十七味丸对非酒精性脂肪性肝炎的作用及作用机制[J]. 中国临床药理学与治疗学, 2026, 31(5): 596-606.

Xiaoyan GAO, Juan GUO, Sheng HUANG, Qiaoyun WANG, Yujia HUANG, Jinyuan LI, Xiaoqiang LIU. Mechanism of Qinggan Ershiqiwei Wan on non-alcoholic steatohepatitis based on network pharmacology and in vivo experiments[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2026, 31(5): 596-606.

图/表 7

图 1

Fig.1 The effect of QGW on body mass index and liver index in HFD-induced NASH mice (x?±s, n=8) A: the initial body weight in each group; B: the trend of body weight within 14 weeks; C: the food intake; D: the consumption of water; E: appearance of the mice liver; F: the liver/weight ratio. cP<0.01, compared with HFD group.

图 2 A-F: the serum level of TG, TC, ALT, AST, IL-1β and IL-6 in n=8 per group. bP<0.05, cP<0.01, compared with HFD group.

Fig.2 The effect of QGW on the levels of TG, TC, ALT, AST, IL-1β, and IL-6 in the serum of HFD-induced NASH mice (x?±s, n=8) A-F: the serum level of TG, TC, ALT, AST, IL-1β and IL-6 in n=8 per group. bP<0.05, cP<0.01, compared with HFD group.

图 3

Fig.3 The effect of QGW on liver tissue morphology in HFD-induced NASH mice (x?±s, n=8) A: H&E staining and Oli&Red staining (magnification 200×); B: quantitative analysis of lipid content; C: quantitative analysis of lipid content of liver tissues. cP<0.01, compare with CON group; fP<0.01, compared with HFD group.

图 4

Fig.4 Network pharmacology analysis of QGW and NASH A: QGW components-disease target Venn diagram; B: network diagram of the active ingredient-target; C: PPI network diagram of QGW and NASH-related targets.

图 5

Fig.5 Network pharmacology analysis of QGW and NASH A-C: GO enrichment analysis; D: KEGG enrichment analysis; E: network diagram of the component-target-pathway.

图 6

Fig.6 The effect of QGW on the MAPK signaling pathway in liver tissues of HFD-induced NASH mice (x?±s, n=8) A: the protein expression levels of p-ERK, ERK, p-JNK, JNK, p-P38, and P38; B-D: quantitative analysis of p-ERK/ERK, p-JNK/JNK and p-P38/P38. bP<0.05, cP<0.01, compared with HFD group.

图 7

Fig.7 The effect of QGW on the distribution of p-ERK, p-JNK, and p-P38 proteins in liver tissues of HFD-induced NASH mice (x?±s, n=8) A: immunohistochemical staining of p-ERK, p-JNK and p-P38, magnification × 200; B-D: the percent of integrated optic density of the area for p-ERK, p-JNK, and p-P38 were analyzed with Image. cP<0.01, compare with CON group; eP<0.05, fP<0.01, compared with HFD group.

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