Objective To explore the effect of microRNA-129-5p (miR-129-5p) on liver injury in metabolic dysfunction-associated fatty liver disease (MAFLD) rats by regulating the Rho kinase (RhoA)/Rho-associated coiled coil kinase (ROCK) pathway. Methods A total of 28 patients with MAFLD who were treated in our hospital from June to December 2024 were selected as MAFLD group, while another 28 healthy individuals undergoing physical examinations were selected as control group. The expression levels of miR-129-5p, RhoA, and ROCK mRNA in the serum of MAFLD group and control group were detected using qRT-PCR method. The dual luciferase reporter gene assay was used to verify the interaction between miR-129-5p and RhoA. MAFLD rats were constructed, and successfully modeled rats were classified into MAFLD group, NC agomir group (tail vein injection of NC agomir), miR-129-5p agomir group (tail vein injection of miR-129-5p agomir), and LPA (tail vein injection of miR-129-5p agomir+1 mg/kg LPA) randomly, each with 12 rats. Another 12 normal rats were considered as NC group. NC group and MAFLD group were injected with equal amounts of physiological saline. The serum liver function, blood lipids, inflammatory factors, and the expression of TC and TG in liver tissue were measured. QRT-PCR was used to detect miR-129-5p, RhoA, and ROCK mRNA in liver tissue of MAFLD rats. HE staining and Oil Red O staining were used to detect liver tissue pathology. TUNEL staining was applied to detect apoptosis of liver tissue cells. Western blot was applied to detect RhoA and ROCK proteins in liver tissue. Results Compared with control group, the expression of miR-129-5p in the serum of MAFLD group decreased, while the expressions of RhoA and ROCK mRNA increased (P<0.05). miR-129-5p could target negative regulation of RhoA. The liver tissue of NC group was normal. The liver tissues of MAFLD group and NC agomir group showed hepatic steatosis, prominent swelling in morphology, balloon like changes, vacuolization, and many red fat droplets. The rats of miR-129-5p agomir group showed a small amount of mild hepatic steatosis, with small circular vacuoles and prominently reduced lipid droplets observed in cytoplasm. The hepatic steatosis in LPA group further worsened, and the number of fat droplets was further increased. The rats in MAFLD group had higher TC in liver and serum, TG, LDL-C, AST, ALT, TNF-α, IL-6, IL-1β, RhoA mRNA and protein, ROCK mRNA and protein in liver and serum, and apoptosis rate than NC group, and lower HDL-C and miR-129-5p than the NC group (P<0.05). The rats in miR-129-5p agomir group had lower TC in liver and serum, TG, LDL-C, AST, ALT, TNF-α, IL-6, IL-1β, RhoA mRNA and protein, ROCK mRNA and protein in the liver and serum, and apoptosis rate than MAFLD group and NC agomir group, and higher HDL-C and miR-129-5p than MAFLD group and NC agomir group (P<0.05). The LPA group had higher TC in liver and serum, TG, LDL-C, AST, ALT, TNF-α, IL-6, IL-1β, RhoA mRNA and protein, ROCK mRNA and protein in liver and serum, and apoptosis rate than miR-129-5p agomir group, and lower HDL-C than miR-129-5p agomir group (P<0.05). Conclusions MiR-129-5p may alleviate liver injury caused by MAFLD by targeting negative regulation of RhoA/ROCK pathway.
Key words
MicroRNA-129-5p /
/
/
Rho-associated coiled-coil kinase /
/
/
Rho kinase /
/
/
Metabolic dysfunction-associated fatty liver disease /
/
/
Liver injury
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
References
[1] Wu L, Mo W, Feng J, et al. Astaxanthin attenuates hepatic damage and mitochondrial dysfunction in non-alcoholic fatty liver disease by up-regulating the FGF21/PGC-1α pathway[J]. Br J Pharmacol, 2020, 177(16):3760-3777. DOI: 10.1111/bph.15099.
[2] Ying Y, Zhang H, Yu D, et al. Gegen Qinlian Decoction Ameliorates Nonalcoholic Fatty Liver Disease in Rats via Oxidative Stress, Inflammation, and the NLRP3 Signal Axis[J]. Evid Based Complement Alternat Med, 2021, 1(1):6659445-6689455. DOI: 10.1155/2021/6659445.
[3] López-Pastor A R, Infante-Menéndez J, Escribano Ó, et al. miRNA Dysregulation in the Development of Non-Alcoholic Fatty Liver Disease and the Related Disorders Type 2 Diabetes Mellitus and Cardiovascular Disease[J]. Front Med (Lausanne), 2020, 7(1):527059-527070. DOI: 10.3389/fmed.2020.527059.
[4] Ye J, Lin Y, Yu Y, et al. LncRNA NEAT1/microRNA-129-5p/SOCS2 axis regulates liver fibrosis in alcoholic steatohepatitis[J]. J Transl Med, 2020, 18(1):445-456. DOI: 10.1186/s12967-020-02577-5.
[5] Hochreuter MY, Dall M, Treebak JT, et al. MicroRNAs in non-alcoholic fatty liver disease: Progress and perspectives[J]. Mol Metab, 2022, 65(1):101581-101597. DOI: 10.1016/j.molmet.2022.101581.
[6] Lu W, Chen Z, Wen J. The role of RhoA/ROCK pathway in the ischemic stroke-induced neuroinflammation[J]. Biomed Pharmacother, 2023, 165(1): 115141-115151. DOI: 10.1016/j.biopha.2023.115141.
[7] Yu W, Gao Y, Zhao Z, et al. Fumigaclavine C ameliorates liver steatosis by attenuating hepatic de novo lipogenesis via modulation of the RhoA/ROCK signaling pathway[J]. BMC Complement Med Ther, 2023, 23(1): 288-298. DOI: 10.1186/s12906-023-04110-9.
[8] 代宇, 范伟. 基于AMPK/mTOR信号通路探究下调miR-34a对非酒精性脂肪肝大鼠的干预效果[J]. 中国老年学杂志, 2024, 44(6):1473-1477. DOI: 10. 3969 / j. issn. 1005-9202.
Dai Y, Fan W. Investigation on the Intervention Effect of Down-regulating miR-34a on Non-alcoholic Fatty liver disease Rats Based on the AMPK/mTOR Signaling Pathway [J]. Chinese Journal of Gerontology, 2024, 44(6): 1473-1477. DOI: 10. 3969 / j. issn. 1005-9202.
[9] 谷孝月, 刘志明, 赵会新, 等. 川陈皮素调节RhoA/ROCK信号通路对妊娠高血压大鼠内皮功能障碍的影响[J]. 中国优生与遗传杂志, 2024, 32(1): 41-47. DOI: 10.13404/j.cnki.cjbhh.2024.01.006.
Gu XY, Liu ZM, Zhao HX, et al. Effect of nobiletin on endothelial dysfunction in pregnancy-induced hypertension rats by regulating RhoA/ROCK signaling pathway[J]. Chinese Journal of Eugenics and Genetics, 2024, 32(1): 41-47. DOI: 10.13404/j.cnki.cjbhh.2024.01.006
[10]Chen M, Xing J, Pan D, et al. Chinese herbal medicine mixture 919 syrup alleviates nonalcoholic fatty liver disease in rats by inhibiting the NF-κB pathway[J]. Biomed Pharmacother, 2020, 128(1):110286-110295. DOI: 10.1016/j.biopha.2020.110286.
[11]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.
[12]Liu XL, Pan Q, Cao HX, et al. Lipotoxic Hepatocyte-Derived Exosomal MicroRNA 192-5p Activates Macrophages Through Rictor/Akt/Forkhead Box Transcription Factor O1 Signaling in Nonalcoholic Fatty Liver Disease[J]. Hepatology, 2020, 72(2): 454-469. DOI: 10.1002/hep.31050.
[13]Reda D, Elshopakey GE, Albukhari TA, et al. Vitamin D3 alleviates nonalcoholic fatty liver disease in rats by inhibiting hepatic oxidative stress and inflammation via the SREBP-1-c/ PPARα-NF-κB/IR-S2 signaling pathway[J]. Front Pharmacol, 2023, 14(1): 1164512-1164524. DOI: 10.3389/fphar.2023.1164512.
[14]Zubáňová V, Červinková Z, Kučera O, et al. The Connection between MicroRNAs from Visceral Adipose Tissue and Non-Alcoholic Fatty Liver Disease[J]. Acta Medica (Hradec Kralove), 2021, 64(1): 1-7. DOI: 10.14712/18059694.2021.1.
[15]Wei Z, Hang S, Wiredu Ocansey D K, et al. Human umbilical cord mesenchymal stem cells derived exosome shuttling mir-129-5p attenuates inflammatory bowel disease by inhibiting ferroptosis[J]. J Nanobiotechnology, 2023, 21(1): 188-206. DOI: 10.1186/s12951-023-01951-x.
[16]Chang J, Zhang Y, Shen N, et al. MiR-129-5p prevents depressive-like behaviors by targeting MAPK1 to suppress inflammation[J]. Exp Brain Res, 2021, 239(11): 3359-3370. DOI: 10.1007/s00221-021-06203-8.
[17]Huang X, Hou X, Chuan L, et al. miR-129-5p alleviates LPS-induced acute kidney injury via targeting HMGB1/TLRs/NF-kappaB pathway[J]. Int Immunopharmacol, 2020, 89(1): 107016. DOI: 10.1016/j.intimp.2020.107016.
[18]Zhu Y, Hu Y, Cheng X, et al. Elevated miR-129-5p attenuates hepatic fibrosis through the NF-κB signaling pathway via PEG3 in a carbon CCl4 rat model[J]. J Mol Histol, 2021, 52(3):491-501. DOI: 10.1007/s10735-020-09949-7.
[19]Zhang R, Li W, Jiang X, et al. Ferulic Acid Combined With Bone Marrow Mesenchymal Stem Cells Attenuates the Activation of Hepatic Stellate Cells and Alleviates Liver Fibrosis[J]. Front Pharmacol, 2022, 13(1): 863797-863813. DOI: 10.3389/fphar.2022.863797.
[20]Rao J, Qiu J, Ni M, et al. Macrophage nuclear factor erythroid 2-related factor 2 deficiency promotes innate immune activation by tissue inhibitor of metalloproteinase 3-mediated RhoA/ROCK pathway in the ischemic liver[J]. Hepatology, 2022, 75(6): 1429-1445. DOI: 10.1002/hep.32184.
[21]Huang Z, Khalifa MO, Gu W, et al. Hydrostatic pressure induces profibrotic properties in hepatic stellate cells via the RhoA/ROCK signaling pathway[J]. FEBS Open Bio, 2022, 12(6): 1230-1240. DOI: 10.1002/2211-5463.13405.
[22]Ren X, Meng T, Ren X, et al. Fasudil alleviates acetaminophen-induced liver injury via targeting Rhoa/ROCK signal pathway[J]. J Toxicol Sci, 2021, 46(6): 255-262. DOI: 10.2131/jts.46.255.
[23]Peng JH, Leng J, Tian HJ, et al. Geniposide and Chlorogenic Acid Combination Ameliorates Non-alcoholic Steatohepatitis Involving the Protection on the Gut Barrier Function in Mouse Induced by High-Fat Diet[J]. Front Pharmacol, 2018, 9(3): 1399-1411. DOI: 10.3389/fphar.2018.01399.
PDF(4147 KB)
