Objective  Based on the network pharmacology and animal experiments, to investigate the protective effect and possible molecular mechanism of ethanol extract from Atractylodes lancea on liver function in mice with liver fibrosis induced by bile duct ligation.

Methods  The main active ingredients atractylodin, atractylenolide I, II and II from Atractylodes lancea were selected, which had been verified by literature and experiments, and the targets of these active ingredients were obtained through the SwissTargetPrediction database. The liver fibrosis disease targets were obtained through On-line Mendelian Inheritance in Man (OMIM), DisGeNET and GeneCards databases. The targets were added to the Wei Sheng Xin platform to find the intersection target for Atractylodes lancea in treating liver fibrosis. Cytoscape 3.10.1 was used to construct the “drug-component-target-disease” network diagram and protein-protein interaction core target network diagram. GO functional enrichment analysis and KEGG pathway  analysis were performed, and molecular docking was performed between active components and core targets. Liver fibrosis was induced in mice by bile duct ligation, and liver function markers were measured.

Results  A total of 91 corresponding targets of atractylodin, atractylenolide I, II and III and 9 296 liver fibrosis disease targets were obtained, including 74 intersecting targets and 31 core targets. KEGG enrichment analysis showed that the main signaling pathways involved included inflammatory pathways such as epidermal growth factor receptor (EGFR) and phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt). Molecular docking results showed that the active ingredients had strong binding activity with the core target protein. The results of animal experiments showed that, compared with the sham surgery group, the model group displayed notable, the liver index, spleen index, activity of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), degree of liver fibrosis, mRNA and protein expression of α-smooth muscle actin (α-SMA) and recombinant collagen type I alpha 1 (COL1A1), and mRNA of recombinant collagen type IV alpha 2 (COL4A2) were significantly increased, and the thymus index was sigficantly decreased (P<0.05); compared with the model group, the liver injury of mice in the Atractylodes lancea administration group reduced liver injury, its liver index, spleen index, activity of serum ALT and AST, degree of liver fibrosis, mRNA and protein expression of α-SMA and COL1A1, and mRNA of COL4A2 were significantly decreased, and the thymus index was sigficantly increased (P<0.05).

Conclusion   Atractylodes lancea can improve liver function and alleviate tissue pathological damage in mice with liver fibrosis, which may be related to activating pathways such as PI3K/Akt, inhibiting oxidative stress and inflammatory reactions, and intervening in liver fibrosis.

1.Chuang CJ, Wu YF, Wu KH, et al. Patients with liver cirrhosis as frequent attenders of emergency departments[J]. Emerg Med Int, 2020, 2020: 8289275. DOI: 10.1155/2020/8289275.

2.Kisseleva T, Brenner D. Molecular and cellular mechanisms of liver fibrosis and its regression[J]. Nat Rev Gastroenterol Hepatol, 2021, 18(3): 151-166. DOI: 10.1038/s41575-020-00372-7.

3.Li H. Advances in anti-hepatic fibrotic therapy with Traditional Chinese Medicine herbal formula[J]. J Ethnopharmacol, 2020, 251: 112442. DOI: 10.1016/j.jep.2019.112442.

4.吴昊, 李梦玲, 邱志海. 五灵胶囊对慢性乙型肝炎患者（肝郁脾虚证）肝纤维化指标及氧化应激反应的影响 [J]. 现代诊断与治疗, 2023, 34(24): 3664-3666. https://d.wanfangdata.com.cn/periodical/xdzdyzl202324011.

5.窦婧, 牛丽娜, 王晓忠. 基于数据挖掘探讨“和法”在肝郁脾虚型肝纤维化治疗中的用药规律[J]. 现代中西医结合杂志, 2024, 33(3): 384-389, 398. DOI: 10.3969/j.issn.1008-8849.2024.03.017.

6.安振涛, 魏兰福, 黄钰萍, 等. 肝纤维化中医证候规范化与客观化研究进展[J]. 中国中医基础医学杂志, 2020, 26(2): 280-282. DOI: 10.3969/j.issn.1006-3250. 2020.02.042.

7.中国药典2020年版. 一部[S]. 2020: 155-168.

8.沙多依, 杭永付, 宋菲, 等. 北苍术炮制前后挥发油部位保肝作用比较研究[J]. 现代中药研究与实践, 2010, 24(4): 41-43. [Sha DY, Hang YF, Song F, et al. Study on the protecting-liver effect of volatile oil of pre and post processing Cangzhu[J]. Research and Practice on Chinese Medicines, 2010, 24(4): 41-43.] DOI: 10.13728/j.1673-6427.2010.04.014.

9.靳露露. 基于真核有参转录组测序分析技术探讨越鞠保和丸改善肝纤维化小鼠抑郁样行为[D]. 天津: 天津中医药大学, 2022. DOI: 10.27368/d.cnki.gtzyy.2022.000288.

10.蔡金阳, 岳天祥, 张月林, 等. 经典名方越鞠丸的历史沿革与处方考证[J]. 中草药, 2023, 54(19): 6511-6519. [Cai JY, Yue TX, Zhang YL, et al. Historical evolution and prescription research of the classic prescription Yueju pill[J]. Chinese Herbal Medicine, 2023, 54(19): 6511-6519.] DOI: 10.7501/j.issn.0253-2670.2023.19.031.

11.Chae HS, Kim YM, Chin YW. Atractylodin inhibits interleukin-6 by blocking NPM-ALK activation and MAPKs in HMC-1[J]. Molecules, 2016, 21(9): 1169. DOI: 10.3390/molecules21091169.

12.Li Q, Tan JX, He Y, et al. Atractylenolide III ameliorates non-alcoholic fatty liver disease by activating hepatic adiponectin receptor 1-mediated AMPK pathway[J]. Int J Biol Sci, 2022, 18(4): 1594-1611. DOI: 10.7150/ijbs.68873.

13.Wang F, Li Z, Chen L, et al. Inhibition of ASCT2 induces hepatic stellate cell senescence with modified proinflammatory secretome through an IL-1α/NF-κB feedback pathway to inhibit liver fibrosis[J]. Acta Pharm Sin B, 2022, 12(9): 3618-3638. DOI: 10.1016/j.apsb.2022.03.014.

14.王倩. 苍术化学成分药理作用研究进展[J]. 生物化工, 2023, 9(1): 158-162. [Wang Q. Research progress on pharmacological action of chemical components of Atractylodes lancea[J]. Biochemistry, 2023, 9(1): 158-162.] DOI: 10.3969/j.issn.2096-0387.2023.01.038.

15.曹阳, 戴国梁, 李斐然, 等. LC-MS/MS测定大鼠血浆中白术内酯Ⅰ、Ⅱ、Ⅲ的浓度及苍术-香附提取物在大鼠体内的药动学研究[J]. 中国药学杂志, 2023, 58(17): 1582-1588. [Cao Y, Dai GL, Li FR, et al. Determination of atractylenolide Ⅰ, Ⅱ and Ⅲ in rat plasma by LC-MS/MS and study on pharmacoki-netics of Atractylodes-Cyperi rhizoma extract in rats[J]. Chinese Journal of Pharmacy, 2023, 58(17): 1582-1588.] DOI: 10.11669/cpj.2023.17.007.

16.Xie Z, Lin M, He X, et al. Chemical constitution, pharmacological effects and the underlying mechanism of atractylenolides: a review[J]. Molecules, 2023, 28(10): 3987. DOI: 10.3390/molecules28103987.

17.Qu L, Liu C, Ke C, et al. Atractylodes lancea rhizoma attenuates DSS-induced colitis by regulating intestinal flora and metabolites[J]. Am J Chin Med, 2022, 50(2): 525-552. DOI: 10.1142/S0192415X22500203.

18.Ke C, Gao J, Tu J, et al. Ganfule capsule alleviates bile duct ligation-induced liver fibrosis in mice by inhibiting glutamine metabolism[J]. Front Pharmacol, 2022, 13: 930785. DOI: 10.3389/fphar.2022.930785.

19.Roehlen N, Crouchet E, Baumert TF. Liver fibrosis: mechanistic concepts and therapeutic perspectives[J]. Cells, 2020, 9(4): 875. DOI: 10.3390/cells9040875.

20.范松松, 曹学峰, 赵荣楠, 等. 胆汁淤积性肝病纤维化机制研究进展[J]. 中国现代医生, 2023, 61(1): 125-128. DOI: 10.3969/j.issn.1673-9701.2023.01.028.

21.杨婷. 双环醇改善小鼠胆总管结扎诱导的胆汁淤积性肝损伤[D]. 天津: 天津医科大学, 2020. DOI: 10.27366/d.cnki.gtyku.2020.001251.

22.侯晓荣, 赵靖, 赵佳, 等. 五味子油联合莪术油防治肝纤维化的作用及机制研究[J]. 中草药, 2022, 53(4): 1059-1067. [Hou XR, Zhao J, Zhao J, et al. Effect and mechanism of Schisandra chinensis oil combined with Zedoary Turmeric oil on liver fibrosis[J]. Chinese Herbal Medicine, 2022, 53(4): 1059-1067.] DOI: 10.7501/j.issn.0253-2670.2022.04.012.

23.Lin WC, Lin WL. Ameliorative effect of Ganoderma lucidum on carbon tetrachloride-induced liver fibrosis in rats[J]. World J Gastroenterol, 2006, 12(2): 265-270. DOI: 10.3748/wjg.v12.i2.265.

24.支廷威, 刘昌辉. 京尼平苷酸通过调控法尼醇X受体对胆管结扎诱导的胆汁淤积小鼠的影响[J]. 中药新药与临床药理, 2022, 33(12): 1606-1613. [Zhi TW, Liu CH. Effect of geniposidic acid on bile duct ligation-induced cholestasis mice through modulation of farnesoid X receptor[J]. New Traditional Chinese Medicine and Clinical Pharmacology, 2022, 33(12): 1606-1613.] DOI: 10.19378/j.issn.1003-9783.2022.12.003.

25.Magee N, Zou A, Ghosh P, et al. Disruption of hepatic small heterodimer partner induces dissociation of steatosis and inflammation in experimental nonalcoholic steatohepatitis[J]. J Biol Chem, 2020, 295(4): 994-1008. DOI: 10.1074/jbc.RA119.010233.

26.刘蓉. 白术内酯Ⅰ调控肝星状细胞自噬及其对肝纤维化的作用机制研究[D]. 成都: 成都医学院, 2023. DOI: 10.27843/d.cnki.gcdyy.2023.000269.

27.Huang K, Du M, Tan X, et al. PARP1-mediated PPARα poly (ADP-ribosyl) ation suppresses fatty acid oxidation in non-alcoholic fatty liver disease[J]. J Hepatol, 2017, 66(5): 962-977. DOI: 10.1016/j.jhep.2016.11.020.

28.Cheng X, Han ZX, Su ZJ, et al. Network pharmacology-based exploration on the intervention of Qinghao Biejia decoction on the inflammation-carcinoma transformation process of chronic liver disease via MAPK and PI3k/AKT pathway[J]. Biomed Res Int, 2022, 2022: 9202128. DOI: 10.1155/2022/9202128.