外源添加或siRNA干扰ACE2对油酸致NAFLD细胞氧化应激和炎性损伤的保护作用及机制

doi:10.3969/j.issn.1674-7968.2025.12.011

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Abstract Nonalcoholic fatty liver disease (NAFLD) is a chronic liver disease characterized by the production of large amounts of lipid deposits in the liver, which poses a serious health risk to humans. In recent years, there has been a gradual increase in the incidence of fatty liver disease in pets and farm animals, which has resulted in serious economic losses to the farming industry. Angiotensin-converting enzyme 2 (ACE2) has been shown to positively regulate anti-inflammatory, anti-injury and anti-fibrotic effects in a variety of diseases, including fatty liver. The previous study of our group found that ACE2 might exert some anti-injury effects on NAFLD induced by exogenous stimuli, but its exact protective mechanism is not clear. In this study, normal rat (Rattus norvegicus) hepatic stromal BRL-3A cells were induced with 0.025~0.200 mmol/L oleic acid (OA) for 24 h to establish a non-alcoholic fatty liver disease (NAFLD) cell model, and hepatocellular injury markers were measured to determine the optimal induction conditions. Subsequently, the cells were treated with either exogenous ACE2 active protein or siRNA targeting ACE2, after which lipid deposition was observed through staining, and changes in parameters including total anti-oxidative capacity (T-AOC), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and inflammatory factors were detected. Western blot detected the protein expression of cellular ACE2, and mas receptor (MasR); ELISA detected the content of Ang Ⅱ and Ang 1-7 in the supernatant of the cells, and clarified the effect of ACE2 on NAFLD cellular damage. Results showed that: 1) NAFLD cell model was successfully established, analyzed by oil red staining to detect the changes of triglyceride (TG) content and reactive oxygen species (ROS) in hepatocytes, and the induction condition of NAFLD cell model was selected to be 0.100 mmol/L oleic acid for 24 h, and 3 different concentrations of OA (low, medium, and high (0.025, 0.100, and 0.200 mmol/L)) were determined for the follow-up test. 2) Compared with the control group, ACE2 protein expression was significantly down-regulated in the high concentration OA-treated group (P<0.05), and there was no significant difference in ACE2 expression in the low- and medium-concentration groups, the expression levels of ACE2 and MasR in the group treated with the addition of ACE2-activated proteins were significantly elevated (P<0.05), and after the transfection of siRNA-ACE2, the expression level of ACE2 was significantly reduced (P<0.05), and MasR expression level was reduced but the difference was not significant; 3) In medium concentration OA treatment, ACE2 and MasR expression was up-regulated, Ang 1-7 and Ang Ⅱ levels were increased, lipid accumulation was increased, oxidative stress was aggravated, antioxidant capacity was weakened, inflammatory factors and NO content were extremely significantly elevated (P<0.01), iNOS activity was extremely significantly enhanced (P<0.01), and anti-inflammatory factor content was extremely significantly reduced (P<0.01); 4) Compared with the OA-treated group, the ACE2 active protein-treated group cellular ACE2, MasR, and Ang 1-7 levels were significantly or extremely significantly up-regulated (P<0.05 or P<0.01) and Ang Ⅱ levels were down-regulated. The relative intracellular lipid droplet area and TG content were significantly reduced (P<0.05), while ALT, AST viability and T-AOC were enhanced to different degrees, and oxidative stress and inflammatory response were reduced.Transfection with siRNA-ACE2 treatment reversed the above changes. The experimental results suggested that treatment with ACE2-activated protein could alleviate cellular lipid deposition, attenuated cellular oxidative stress and inflammatory response, and had a certain alleviating effect on OA-induced NAFLD in rat hepatocytes. The mechanism was mainly realized by degrading Ang Ⅱ, activating Ang 1-7/Mas, and acting on the NO/iNOS signaling pathway. Thus, this study revealed a mechanism by which ACE2 alleviates NAFLD at the cellular level and provide an important theoretical foundation for developing ACE2 as a potential therapeutic target for NAFLD.

Key words： Angiotensin converting enzyme 2 (ACE2) Nonalcoholic fatty liver disease (NAFLD) Small interfering RNA (siRNA) interference Oxidative stress Inflammatory response

Received: 24 February 2025

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Corresponding Authors: * Corresponding author, zhangyuanshu@njau.edu.cn

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	XU Jia-Jing
	WANG Huan-Huan
	MA Chang
	CHEN Xi-Wen
	WANG Gong-Min
	LIU Ying
	ZHANG Yuan-Shu

Cite this article:

XU Jia-Jing,WANG Huan-Huan,MA Chang, et al. Protective Effects and Mechanisms of Exogenous Addition or siRNA Interference with ACE2 on Oxidative Stress and Inflammatory Injury in NAFLD Cells Caused by Oleic Acid[J]. 农业生物技术学报, 2025, 33(12): 2677-2690.

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https://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2025.12.011 OR https://journal05.magtech.org.cn/Jwk_ny/EN/Y2025/V33/I12/2677

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