Objective  To investigate the effect of Asiaticoside (ASI) on liver injury in non-alcoholic fatty liver disease (NAFLD) and its impacts on the microRNA-29b-3p (miR-29b-3p)/forkhead box protein O3 (FoxO3) axis.

Methods  Rats were stochastically grouped into a Control group, a Model group, a low-dose ASI-L group (ASI-L), a high-dose ASI-H group (ASI-H), a high-dose ASI+blank vector group (ASI-H+Vector), and a high-dose ASI+miR-29b-3p over expression group (ASI-H+OV-miR-29b-3p), with 12 rats in each group. The changes in rat liver index were analyzed. HE staining method was applied to observe the pathological changes of liver tissue. Biochemical analyzer was used to detect lipid metabolism indicators and indicators reflecting liver parenchymal damage in rats. The levels of oxidative stress and inflammatory cytokines were detected by the ELISA method. The RT-qPCR method was applied to detect the mRNA levels of miR-29b-3p and FoxO3 in rat liver tissue. The Western blot method was applied to detect the expression levels of FoxO3 and apoptosis-related proteins in rat liver tissue. Dual luciferase activity was applied to validate the targeting relationship between miR-29b-3p and FoxO3.

Results  Compared with the Control group, rats in the Model group showed severe liver tissue damage, liver cell rupture, and extensive infiltration of inflammatory cells, the liver weight and liver index increased, the levels of TC, TG, AST, ALT, MDA, TNF-α, IL-6 obviously increased, the levels of SOD and CAT obviously reduced, the expression level of miR-29b-3p and expression level of Caspase 3 protein in liver tissue obviously increased, the expression level of FoxO3 mRNA and expression levels of FoxO3, Bcl-2 proteins decreased (P<0.05). Compared with the model group, as the dose of ASI increased, the degree of liver tissue damage, liver cell rupture, and inflammatory cell infiltration of rats in the ASI-L and ASI-H groups decreased, the liver weight and liver index gradually decreased, the levels of TC, TG, AST, ALT, MDA, TNF-α, IL-6 in serum gradually decreased, the levels of SOD and CAT gradually increased, the expression level of miR-29b-3p and expression level of Caspase 3 protein in liver tissue gradually decreased, the expression level of FoxO3 mRNA and the expression levels of FoxO3 and Bcl-2 proteins gradually increased. miR-29b-3p overexpression reversed the improvement effect of ASI on liver tissue in NAFLD rats. Dual luciferase activity confirmed the targeting relationship between miR-29b-3p and FoxO3.

Conclusion  ASI can enhance the antioxidant function, regulate lipid metabolism, alleviate inflammation and liver tissue damage in NAFLD rats, which may be related to the down-regulation of miR-29b-3p and the up-regulation of FoxO3 expression.

1.Rong L, Zou J, Ran W, et al. Advancements in the treatment of non-alcoholic fatty liver disease (NAFLD)[J]. Front Endocrinol (Lausanne), 2023, 13(1): 1-18. DOI: 10.3389/fendo.2022.1087260.

2.Guo X, Yin X, Liu Z, et al. Non-alcoholic fatty liver disease (NAFLD) pathogenesis and natural products for prevention and treatment[J]. Int J Mol Sci, 2022, 23(24): 15489-15506. DOI: 10.3390/ijms232415489.

3.Raza S, Rajak S, Upadhyay A, et al. Current treatment paradigms and emerging therapies for NAFLD/NASH[J]. Front Biosci (Landmark Ed), 2021, 26(2): 206-237. DOI: 10.2741/4892.

4.Fan L, Li X, Liu T. Asiaticoside inhibits neuronal apoptosis and promotes functional recovery after spinal cord injury in rats[J]. J Mol Neurosci, 2020, 70(12): 1988-1996. DOI: 10.1007/s12031-020-01601-z.

5.Guo M, Xu J, Wang S, et al. Asiaticoside reduces autophagy and improves memory in a rat model of dementia through mTOR signaling pathway regulation[J]. Mol Med Rep, 2021, 24(3): 645-656. DOI: 10.3892/mmr.2021.12284.

6.Xiao X, Zhang Q. Asiaticoside conveys an antifibrotic effect by inhibiting activation of hepatic stellate cells via the Jagged-1/Notch-1 pathway[J]. J Nat Med, 2023, 77(1): 128-136. DOI: 10.1007/s11418-022-01653-y.

7.Nie J, Li CP, Li JH, et al. Analysis of nonalcoholic fatty liver disease microRNA expression spectra in rat liver tissues[J]. Mol Med Rep, 2018, 18(3): 2669-2680. DOI: 10.3892/mmr.2018.9268.

8.Dalgaard LT, Sørensen AE, Hardikar AA, et al. The microRNA-29 family: role in metabolism and metabolic disease[J]. Am J Physiol Cell Physiol, 2022, 323(2): C367-C377. DOI: 10.1152/ajpcell.00051.2022.

9.Wang L, Zhu X, Sun X, et al. FoxO3 regulates hepatic triglyceride metabolism via modulation of the expression of sterol regulatory-element binding protein 1c[J]. Lipids Health Dis, 2019, 18(1): 197-208. DOI: 10.1186/s12944-019-1132-2.

10.杨仁国, 罗婷婷, 贺微微. miR-29b-3p靶向FoxO3基因改善非酒精性脂肪肝预后的初步研究[J]. 重庆医科大学学报, 2022, 47(9): 1005-1011. [Yang RG, Luo  TT, He WW. miR- 29b- 3p regulates lipid metabolism and fibrosis by targeting IGF-1 in non-alcoholic fatty liver disease[J]. Journal of Chongqing Medical University, 2022, 47(9): 1005-1011.] DOI: 10. 3969 / j.issn.1671-7856.

11.Wang S, Sheng F, Zou L, et al. Hyperoside attenuates non-alcoholic fatty liver disease in rats via cholesterol metabolism and bile acid metabolism[J]. J Adv Res, 2021, 34(1): 109-122. DOI: 10.1016/j.jare.2021.06.001.

12.孙红爽, 李鹏霖, 刘永双, 等. 大黄素对大鼠非酒精性脂肪肝及肝组织11β-羟基类固醇脱氢酶1表达的影响[J]. 实验动物科学, 2022, 39(3): 22-26, 32. [Sun HS, Li PL, Liu YS, et  al. Effects of emodin on the expression of 11β-hydroxy steroid dehydrogenase 1 in nonalcoholic fatty liver and liver tissue of rats[J]. Journal of Experimental Animal Science, 2022, 39(3): 22-26, 32.] DOI: 10.3969/j.issn.1006-6179.2022.03.005.

13.王迎迎, 顾永政. 积雪草苷对代谢相关脂肪性肝病大鼠Nrf2、ROS通路表达的影响[J]. 现代药物与临床, 2024, 39(4): 810-815. [Wang YY, Gu YZ. Effects of asiaticoside on the expression of Nrf2 and ROS pathways in rats with metabolism-related fatty liver disease[J]. Modern Medicine and Clinic, 2024, 39(4): 810-815.] DOI: 10.7501/j.issn.1674-5515.2024.04.002.

14.Liu X, Liu T, Yang K, et al. Antifatigue Effect of asiaticoside in mice by attenuating oxidative stress[J]. Discov Med, 2023, 35(176): 275-282. DOI: 10.24976/Discov.Med.202335176.28.

15.Ma Y, Wen J, Wang J, et al. Asiaticoside antagonizes proliferation and chemotherapeutic drug resistance in hepatocellular carcinoma (HCC) Cells[J]. Med Sci Monit, 2020, 26(1): 1-11. DOI: 10.12659/MSM.924435.

16.Wei Y, Zhang Y, Zhan B, et al. Asiaticoside alleviated NAFLD by activating Nrf2 and inhibiting the NF-κB pathway[J]. Phytomedicine, 2025, 136(1): 1-12. DOI: 10.1016/j.phymed.2024.156317.

17.Tamber SS, Bansal P, Sharma S, et al. Biomarkers of liver diseases[J]. Mol Biol Rep, 2023, 50(9): 7815-7823. DOI: 10.1007/s11033-023-08666-0.

18.Yang K, Chen J, Zhang T, et al. Efficacy and safety of dietary polyphenol supplementation in the treatment of non-alcoholic fatty liver disease: a systematic review and Meta-analysis[J]. Front Immunol, 2022, 13(1): 1-21. DOI: 10.3389/fimmu.2022.949746.

19.Xu C, Xia L, Xu D, et al. Cardioprotective effects of asiaticoside against diabetic cardiomyopathy: activation of the AMPK/Nrf2 pathway[J]. J Cell Mol Med, 2024, 28(2): e18055-e18068. DOI: 10.1111/jcmm.18055.

20.Zhang C, Ji X, Chen Z, et al. Asiaticoside suppresses gastric cancer progression and induces endoplasmic reticulum stress through the miR-635/HMGA1 axis[J]. J Immunol Res, 2022, 2022(1): 1-12. DOI: 10.1155/2022/1917585.

21.Malladi N, Lahamge D, Somwanshi BS, et al. Paricalcitol attenuates oxidative stress and inflammatory response in the liver of NAFLD rats by regulating FOXO3a and NFκB acetylation[J]. Cell Signal, 2024, 121(1): 1-11. DOI: 10.1016/j.cellsig.2024.111299.

22.Zhou J, Li W, Chi X, et al. Inhibition of mmu_circ_0009303 improves metabolic dysfunction-associated steatotic liver disease by regulating lipid metabolism and oxidative stress[J]. Endocr J, 2025, 72(1): 79-91. DOI: 10.1507/endocrj.EJ24-0008.

23.Hu Y, Peng X, Du G, et al. MicroRNA-122-5p inhibition improves inflammation and oxidative stress damage in dietary-induced non-alcoholic fatty liver disease through targeting FOXO3[J]. Front Physiol, 2022, 13(1): 1-12. DOI: 10.3389/fphys.2022.803445.

24.支晓雯, 李兵. 积雪草苷在高糖诱导人角膜上皮细胞损伤修复中的作用及机制研究[J]. 眼科新进展, 2024, 44(5): 360-364. [Zhi XW, Li B. Study on the role and mechanism of asiaticoside in the repair of human corneal epithelial cell damage induced by high glucose[J]. Recent Advances in Ophthalmology, 2024, 44(5): 360-364.] DOI: 10.13389/j.cnki.rao.2024.0070.

25.Fiorucci S, Di Giorgio C, Distrutti E. Obeticholic acid: an update of its pharmacological activities in liver disorders[J]. Handb Exp Pharmacol, 2019, 256(1): 283-295. DOI: 10.1007/164_2019_227.