Xie Zhuo Tiao Zhi formula ameliorates chronic alcohol-induced liver injury in mice

Abstract

This study aimed to evaluate the protective role and potential mechanisms of Xie Zhuo Tiao Zhi decoction (XZTZ) on alcohol-associated liver disease (ALD). XZTZ significantly alleviated alcohol-induced liver dysfunction, based on histological examinations and biochemical parameters after 4-week administration. Mechanically, alcohol-stimulated hepatic oxidative stress was ameliorated by XZTZ, accompanied by the improvement of Nrf2/Keap1 expression and alcohol-activated phosphorylation of pro-inflammatory transcription factors, including JNK, P38, P65, and IκBα, were rescued by XZTZ. In conclusion, XZTZ demonstrates potential in alleviating alcohol-induced liver injury, oxidative stress, and inflammation possibly through modulation of Nrf2/Keap1 and MAPKs/NF-κB signaling pathways, suggesting its potential as a therapeutic option for patients with alcoholic liver disease.

Keywords: Xie Zhuo Tiao Zhi decoction; alcohol-associated liver disease; liver inflammation; liver injury; oxidative stress.

FIGURE 1

The XZTZ rescues alcohol-induced liver injury in C57BL/6J mice. (A) The experimental design graphic; (B, C) Serum levels of ALT and AST; (D) H&E stained sections of the liver (×200 and ×400); (E) Western blot verifying the effects of XZTZ on expression of cleaved-Caspase3 and Bcl-2 with alcohol treatment. ns, not significant; *p < 0.05, **p < 0.01 and ***p < 0.001 indicate statistically significant differences.

FIGURE 2

The XZTZ alleviates alcohol-induced hepatic oxidative stress in C57BL/6J mice. (A) The level of MDA in mice livers; (B–D) The activities of SOD, CAT, and GSH-Px in mice livers; (E) The protein level of CYP2E1 in the liver of mice. ns, not significant; *p < 0.05, **p < 0.01 and ***p < 0.001 indicate statistically significant differences.

FIGURE 3

The XZTZ inhibits hepatic oxidative stress via activating the Nrf2-Keap1 pathway. (A) Western blot verifying the nuclear protein level of Nrf2 and the total protein level of Keap1. LaminB is used as an internal control for nuclear protein and β-actin is used as an internal control for total protein; (B) qRT-PCR verifying the expression levels of HO-1, GCLC, and GPX-1 mRNAs. ns, not significant; *p < 0.05, **p < 0.01 and ***p < 0.001 indicate statistically significant differences.

FIGURE 4

The XZTZ improves alcohol-induced liver inflammation in C57BL/6J mice. (A) Representative stainings of MPO-positive cells in mice livers. Scale bars: 200μM; (B) Gene expression levels of hepatic TNFA, IL-6, IL-1B, and MCP-1. *p < 0.05, **p < 0.01 and ***p < 0.001 indicate statistically significant differences.

FIGURE 5

The XZTZ eases hepatic inflammatory response via MAPKs/NF-kB pathways. (A) The expression of hepatic p-JNK/JNK and p-P38/P38; (B) The expression of hepatic p-P65/P65 and p-IκBα/IκBα. ns, not significant; *p < 0.05 and **p < 0.01 indicate statistically significant differences.

FIGURE 6

Schematic diagram of our study.