The effect of perindopril on the NOX4/NLRP3 signaling pathway in TAA-induced liver fibrosis in rats

doi:10.11958/20252146

Abstract

Abstract:

Objective To investigate the effect of perindopril on the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 4 (NOX4)/NOD-like receptor protein 3 (NLRP3) signaling pathway in rat liver fibrosis induced by thioacetamide (TAA). Methods Thirty-two SD rats were randomly divided into the blank group, the model group (TAA 200 mg/kg, twice a week for 6 weeks) and the low/high dose group (TAA+ perindopril 2/8 mg/kg), with 8 rats in each group. Two weeks after modeling, the administration group was given the corresponding dose of perindopril by gavage (for 4 weeks). At the end of the 6th week, liver pathological sections were used to observe pathological changes of liver tissue and degrees of fibrosis and inflammation. The biochemical analyzer was used to detect alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Enzyme-linked immunosorbent assay (ELISA) was used to detect NLRP3 and IL-1β. Immunohistochemistry was used to detect NOX4, NLRP3, Caspase-1, IL-1β protein and α -smooth muscle agonist protein (α-SMA). Results Compared with the blank group, liver collagen fibers were significantly proliferated in the model group, a large number of inflammatory cells infiltrated, and serum levels of ALT and AST, as well as NLRP3 and IL-1β in rats were significantly increased. The average optical density values of positive proteins such as NOX4, NLRP3, Caspase-1, IL-1β and α-SMA in rat liver tissue increased significantly (P < 0.05). Compared with the model group, the proliferation of collagen fibers and inflammatory infiltration were significantly reduced in both the low-dose and high-dose groups, and serum levels of ALT and AST, NLRP3 and IL-1β in rats were significantly decreased. The average optical density values of positive proteins such as NOX4, NLRP3, Caspase-1, IL-1β and α-SMA in rat liver tissue decreased significantly (P < 0.05). Moreover, the degree of liver fibrosis reduction in rats was better in the high-dose group than that in the low-dose group. Conclusion Perindopril may regulate the NLRP3 signaling pathway by inhibiting the expression of NOX4, thereby reducing oxidative stress damage and inflammatory responses and thus delaying the process of liver fibrosis.

Key words: hepatic fibrosis, perindopril, NADPH oxidase 4, NLR family, pyrin domain-containing 3 protein, oxidative stress, inflammation

CLC Number:

R575.2

Figures/Tables 5

References 23

[1]	KONG Y, CHEN Z, NIE Z, et al. Targeting endoplasmic reticulum proteostasis in liver fibrosis:from signaling mechanisms to therapeutic opportunities[J]. Pharmacol Res, 2025, 217:107823. doi:10.1016/j.phrs.2025.107823.
[2]	CHARAN H V, DWIVEDI D K, KHAN S, et al. Mechanisms of NLRP3 inflammasome-mediated hepatic stellate cell activation:therapeutic potential for liver fibrosis[J]. Genes Dis, 2023, 10(2):480-494. doi:10.1016/j.gendis.2021.12.006.
[3]	OSMAN H A, ABUHAMDAH S, HASSAN M H, et al. NLRP3 inflammasome pathway involved in the pathogenesis of metabolic associated fatty liver disease[J]. Sci Rep, 2024, 14(1):19648. doi:10.1038/s41598-024-69764-y.
[4]	ZHAI L, PEI H, YANG Y, et al. NOX4 promotes Kupffer cell inflammatory response via ROS-NLRP3 to aggravate liver inflammatory injury in acute liver injury[J]. Aging (Albany NY), 2022, 14(17):6905-6916. doi:10.18632/aging.204173.
[5]	NG W H, YEO Y H, KIM H, et al. Renin-angiotensin-aldosterone system inhibitor use improves clinical outcomes in patients with metabolic dysfunction-associated steatotic liver diseases:target trial emulation using real-world data[J]. Hepatology, 2025. doi:10.1097/HEP.0000000000001333. [Epub ahead of print ].
[6]	ZHANG X, WONG G L, YIP T C, et al. Angiotensin-converting enzyme inhibitors prevent liver-related events in nonalcoholic fatty liver disease[J]. Hepatology, 2022, 76(2):469-482. doi:10.1002/hep.32294.
[7]	REZA H M, TABASSUM N, SAGOR M A, et al. Angiotensin-converting enzyme inhibitor prevents oxidative stress,inflammation,and fibrosis in carbon tetrachloride-treated rat liver[J]. Toxicol Mech Methods, 2016, 26(1):46-53. doi:10.3109/15376516.2015.1124956.
[8]	LIANG H, ZHANG N, ZHAO L, et al. Perindopril improves cardiac fibrosis through targeting the AngII/AT1R pathway[J]. Cell Mol Biol (Noisy-le-grand), 2023, 69(9):234-238. doi:10.14715/cmb/2023.69.9.36.
[9]	ABD EL-RAHMAN S S. FAYED H M. Targeting AngII/AT1R signaling pathway by perindopril inhibits ongoing liver fibrosis in rat[J]. J Tissue Eng Regen Med, 2019, 13(12):2131-2141. doi:10.1002/term.2940.
[10]	LIU J, BAI Y, YAO W, et al. Using intra-voxel incoherent motion MRI to dynamically evaluate the attenuating effects of donafenib combined with carvedilol in a thioacetamide-induced hepatic fibrosis rat model[J]. MAGMA, 2025. doi:10.1007/s10334-025-01241-7. [Online ahead of print].
[11]	HAO X, XU L, LAN X, et al. Impact of hepatic inflammation and fibrosis on the recurrence and long-term survival of hepatitis B virus-related hepatocellular carcinoma patients after hepatectomy[J]. BMC Cancer, 2024, 24(1):475. doi:10.1186/s12885-024-12187-9.
[12]	RAMOS-TOVAR E, MURIEL P. Molecular mechanisms that link oxidative stress,inflammation,and fibrosis in the liver[J]. Antioxidants (Basel), 2020, 9(12):1279. doi:10.3390/antiox9121279.
[13]	MEGAHED A, GADALLA H, ABDELHAMID F M, et al. Vitamin D ameliorates the hepatic oxidative damage and fibrotic effect caused by thioacetamide in rats[J]. Biomedicines, 2023, 11(2):424. doi:10.3390/biomedicines11020424.
[14]	BIN ATAN N, BIN HADI MF, TEOH V, et al. ARNI versus perindopril for remodeling in HFrEF. A cohort study[J]. J Cardiovasc Pharmacol Ther, 2023,28:10742484231195019. doi:10.1177/10742484231195019.
[15]	TOPCU A, KOSTAKOGLU U, MERCANTEPE T, et al. The cardioprotective effects of perindopril in a model of polymicrobial sepsis:the role of radical oxygen species and the inflammation pathway[J]. J Biochem Mol Toxicol, 2022, 36(8):e23080. doi:10.1002/jbt.23080.
[16]	FENG J, HUANG X, XU Q, et al. Pharmacological inhibition of the ACE/Ang-2/AT1 axis alleviates mechanical ventilation-induced pulmonary fibrosis[J]. Int Immunopharmacol, 2024, 131:111855. doi:10.1016/j.intimp.2024.111855.
[17]	KAMEL E O, HASSANEIN E, AHMED M A, et al. Perindopril ameliorates hepatic ischemia reperfusion injury via regulation of NF-κB-p65/TLR-4,JAK1/STAT-3,Nrf-2,and PI3K/Akt/mTOR signaling pathways[J]. Anat Rec (Hoboken), 2020, 303(7):1935-1949. doi:10.1002/ar.24292.
[18]	李静, 申风俊. 贝那普利对肝纤维化大鼠NOX4和Nrf2的影响[J]. 山西医科大学学报, 2022, 53(5):573-577.
	LI J, SHEN F J. Effect of benazepril on NOX4 and Nrf2 in rats with hepatic fibrosis[J]. J Shanxi Med Univ, 2022, 53(5):573-577. doi:10.13753/j.issn.1007-6611.2022.05.010.
[19]	陈晨晨, 宋珂, 丁樱, 等. 雷公藤多苷对IgA肾病大鼠肾脏保护及肾组织CB2R/NOX4/NLRP3表达的影响[J]. 中国药理学通报, 2024, 40(11):2037-2041.
	CHEN C C, SONG K, DING Y, et al. The effect of Tripterygium glycosides on renal protection and the expression of CB2R/NOX4/NLRP3 in renal tissue of rats with IgA nephropathy[J]. Chinese Journal of Pharmacology, 2024, 40(11):2037-2041. doi:10.12360/CPB202405051.
[20]	ZHU L, TONG H, REN C, et al. Inflammation unleashed:the role of pyroptosis in chronic liver diseases[J]. Int Immunopharmacol, 2024, 141:113006. doi:10.1016/j.intimp.2024.113006.
[21]	冯浩英华, 房广庆, 严玥, 等. 益气活血固肾颗粒通过调节Nox4抑制炎症和氧化应激改善糖尿病肾病肾脏纤维化[J]. 中药新药与临床药理, 2025, 36(6):888-898.
	FENG H Y H, FANG G Q, YAN Y, et al. Yiqi Huoxue Gushen Granules improve renal fibrosis in diabetic nephropathy by regulating Nox4 to inhibit inflammation and oxidative stress[J]. Clinical Pharmacology and New Drug of Chinese Medicine, 2025, 36(6):888-898. doi:10.19378/j.issn.1003-9783.2025.06.006.
[22]	NIE Y, LIU Q, ZHANG W, et al. Ursolic acid reverses liver fibrosis by inhibiting NOX4/NLRP3 inflammasome pathways and bacterial dysbiosis[J]. Gut Microbes, 2021, 13(1):1972746. doi:10.1080/19490976.2021.1972746.
[23]	XU W, SONG S, HUANG Y, et al. Effects of perindopril and valsartan on expression of transforming growth factor-beta-Smads in experimental hepatic fibrosis in rats[J]. J Gastroenterol Hepatol, 2006, 21(8):1250-1256. doi:10.1111/j.1440-1746.2006.04331.x.

组别	ALT	AST
空白组	53.63±7.33	152.88±17.11
模型组	148.63±23.79^a	386.13±35.41^a
低剂量组	82.25±9.45^b	194.25±23.03^b
高剂量组	64.75±9.82^bc	168.50±13.65^bc
F	71.886^＊＊	166.365^＊＊

组别	ALT	AST
空白组	53.63±7.33	152.88±17.11
模型组	148.63±23.79^a	386.13±35.41^a
低剂量组	82.25±9.45^b	194.25±23.03^b
高剂量组	64.75±9.82^bc	168.50±13.65^bc
F	71.886^＊＊	166.365^＊＊

组别	NLRP3	IL-1β
空白组	3.67±0.36	26.59±5.40
模型组	6.52±0.54^a	63.70±10.70^a
低剂量组	5.74±0.75^b	53.30±8.21^b
高剂量组	4.27±0.68^bc	35.35±5.00^bc
F	38.186^＊＊	38.412^＊＊

组别	NLRP3	IL-1β
空白组	3.67±0.36	26.59±5.40
模型组	6.52±0.54^a	63.70±10.70^a
低剂量组	5.74±0.75^b	53.30±8.21^b
高剂量组	4.27±0.68^bc	35.35±5.00^bc
F	38.186^＊＊	38.412^＊＊

组别	α-SMA	NOX4	NLRP3	Caspase-1	IL-1β
空白组	0.136±0.007	0.048（0.044，0.050）	0.146（0.134，0.147）	0.000（0.000，0.040）	0.064（0.059，0.073）
模型组	0.308±0.012^a	0.311（0.298，0.321）^a	0.318（0.314，0.339）^a	0.259（0.223，0.315）^a	0.325（0.320，0.333）^a
低剂量组	0.242±0.008^b	0.179（0.148，0.189）^b	0.276（0.274，0.288）^b	0.176（0.175，0.188）^b	0.286（0.199，0.290）^b
高剂量组	0.183±0.007^bc	0.081（0.079，0.101）^bc	0.168（0.165，0.179）^bc	0.133（0.125，0.137）^bc	0.123（0.096，0.134）^bc
F或H	620.748^＊＊	28.946^＊＊	32.838^＊＊	32.986^＊＊	32.838^＊＊