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. 2022 Jul 18:13:915193.
doi: 10.3389/fphys.2022.915193. eCollection 2022.

SIRT3 inhibitor 3-TYP exacerbates thioacetamide-induced hepatic injury in mice

Chunxia Shi  1 , Fangzhou Jiao  2 , Yao Wang  1 , Qian Chen  1 , Luwen Wang  1 , Zuojiong Gong  1
Affiliations

SIRT3 inhibitor 3-TYP exacerbates thioacetamide-induced hepatic injury in mice

Chunxia Shi et al. Front Physiol. .

Abstract

The purpose of the study was to explore the effects of SIRT3 inhibitor 3-TYP on acute liver failure (ALF) in mice and its underlying mechanism. The mice were treated with thioacetamide (TAA, 300 mg/kg) for inducing ALF model. 3-TYP (50 mg/kg) was administered 2 h prior to TAA. The liver histological changes were measured by HE staining. Blood samples were collected for analysis of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). MDA and GSH were used to evaluate the oxidative stress of liver. The expression levels of inflammatory cytokines (TNF-α and IL-1β) were measured by ELISA and Western blotting. The cell type expression of IL-1β in liver tissue was detected by immunofluorescent staining. The expression of SIRT3, MnSOD, ALDH2, MAPK, NF-κB, Nrf2/HO-1, p-elF2α/CHOP, and cleaved caspase 3 was determined by Western blotting. TUNEL staining was performed to detect the apoptosis cells of liver tissues. 3-TYP exacerbated the liver injury of ALF mice. 3-TYP increased the inflammatory responses and activation of MAPK and NF-κB pathways. In addition, 3-TYP administration enhanced the damage of oxidative stress, endoplasmic reticulum stress, and promoted hepatocyte apoptosis in ALF mice. 3-TYP exacerbates thioacetamide-induced hepatic injury in mice. Activation of SIRT3 could be a promising target for the treatment of ALF.

Keywords: 3-TYP; SIRT3; acute liver failure; inflammation; thioacetamide.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Effect of 3-TYP on liver damage and survival rate in each group, n = 10 per group. (A) The gross morphology of liver tissues. (B) HE staining of liver tissues. (C) serum ALT levels. (D) serum AST levels. (E) The 24 h survival rates of mice. *p < 0.05, vs. the control group. #p < 0.05, vs. the TAA group.
FIGURE 2
FIGURE 2
Effect of 3-TYP on inflammatory response in each group, n = 10 per group. (A) liver contents of GSH in liver tissues. (B) the MDA level in liver tissues. (C) the serum IL-1β levels. (D) the serum TNF-α levels. (E) The protein expressions of IL-1β and TNF-α in liver tissues were analyzed by western blotting. (F) The quantitative values of IL-1β. (G) The quantitative values of TNF-α. (H) The protein expressions of SIRT3, MnSOD, and ALDH2 in liver tissues were analyzed by western blotting. (I) The quantitative values of SIRT3. (J) The quantitative values of MnSOD. (K) The quantitative values of ALDH2. *p < 0.05, vs. the control group. #p < 0.05, vs. the TAA group.
FIGURE 3
FIGURE 3
Effect of 3-TYP on the macrophage expression of IL-1β in each group, n = 10 per group. (A) Double immunofluorescent staining with macrophage marker anti-CD68 (green) and anti-IL-1β (red). (B) the quantitative co-expressions of CD68 and IL-1β. *p < 0.05, vs. the control group. #p < 0.05, vs. the TAA group. → the co-localization of CD68 and IL-1β.
FIGURE 4
FIGURE 4
Effect of 3-TYP on activation of MAPK and NF-κB pathways in each group, n = 10 per group. (A) The expressions of p-P38, P38, p-ERK1/2, ERK1/2, p-JNK, and JNK in liver tissues were detected by western blotting. (B) The quantitative values of p-P38/P38. (C) The quantitative values of p-ERK/ERK. (D) The quantitative blots of p-JNK/JNK. (E) The quantitative blots of p-P65/P65. *p < 0.05, vs. the control group. #p < 0.05, vs. the TAA group.
FIGURE 5
FIGURE 5
Effect of 3-TYP on activation of oxidative stress and endoplasmic reticulum stress in each group, n = 10 per group. (A) The expressions of Nrf2 and HO-1 in liver tissues were analyzed by western blotting. (B) The quantitative blots of Nrf2. (C) The quantitative blots of HO-1. (D) The expressions of p-elF2α, elF2α, and CHOP in liver tissues were measured by western blotting. (E) The quantitative values of p-elF2α/elF2α. (F) The quantitative values of CHOP. *p < 0.05, vs. the control group. #p < 0.05, vs. the TAA group.
FIGURE 6
FIGURE 6
Effects of 3-TYP on liver cell apoptosis in each group, n = 10 per group. (A) The apoptosis cells of liver samples were measured by TUNEL staining. (B) Quantification of apoptosis. (C) The expressions of cleaved caspase 3 were detected by western blotting. (D) The quantitative values of cleaved caspase 3. *p < 0.05, vs. the control group. #p < 0.05, vs. the TAA group.
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