Recombinant Schistosoma japonicum cystatin alleviates acute liver injury in mice by inhibiting endoplasmic reticulum stress, inflammation and hepatocyte apoptosis

doi:10.12122/j.issn.1673-4254.2024.06.13

Abstract

Abstract:

Objective To investigate the protective effect of recombinant Schistosoma japonicum cystatin (rSj-Cys) against acute liver injury induced by lipopolysaccharide (LPS) and D-GalN in mice. Methods Adult male C57BL/6J mice with or without LPS/D-GaIN-induced acute liver injury were given intraperitoneal injections of rSj-Cys or PBS 30 min after modeling (n=18), and serum and liver tissues samples were collected from 8 mice in each group 6 h after modeling. The survival of the remaining 10 mice in each group within 24 h was observed. Serum levels of ALT, AST, TNF‑α and IL-6 of the mice were measured, and liver pathologies was observed with HE staining. The hepatic expressions of macrophage marker CD68, Bax, Bcl-2 and endoplasmic reticulum stress (ERS)-related proteins were detected using immunohistochemistry or immunoblotting, and TUNEL staining was used to detect hepatocyte apoptosis. Results The survival rates of PBS- and rSj-Cys-treated mouse models of acute liver injury were 30% and 80% at 12 h and were 10% and 60% at 24 h after modeling, respectively; no death occurred in the two control groups within 24 h. The mouse models showed significantly increased serum levels of AST, ALT, IL-6 and TNF-α and serious liver pathologies with increased hepatic expressions of CD68 and Bax, lowered expression of Bcl-2, increased hepatocyte apoptosis, and up-regulated expressions of ERS-related signaling pathway proteins GRP78, CHOP and NF-κB p-p65. Treatment of the mouse models significantly lowered the levels of AST, ALT, IL-6 and TNF‑α, alleviated liver pathologies, reduced hepatic expressions of CD68, Bax, GRP78, CHOP and NF‑κB p-p65, and enhanced the expression of Bcl-2. In the normal control mice, rSj-Cys injection did not produce any significant changes in these parameters compared with PBS. Conclusion rSj-Cys alleviates LPS/D-GalN-induced acute liver injury in mice by suppressing ERS, attenuating inflammation and inhibiting hepatocyte apoptosis.

Key words: acute liver injury, Schistosoma japonicum cystatin, endoplasmic reticulum stress, inflammatory response, apoptosis

Lingjun LU, Xiaodi YANG, Huaping ZHANG, Yuan LIANG, Xiulan SHI, Xin ZHOU. Recombinant Schistosoma japonicum cystatin alleviates acute liver injury in mice by inhibiting endoplasmic reticulum stress, inflammation and hepatocyte apoptosis[J]. Journal of Southern Medical University, 2024, 44(6): 1126-1134.

Figures/Tables 8

Fig.1 Effect of rSj-Cys on survival of the mice after acute liver injury.

Fig.2 Effects of rSj-Cys on serum ALT and AST levels in mice with acute liver injury. ##P<0.01 vs Control group; **P<0.01 vs LPS/D-GaIN group.

Fig.3 Effect of rSj-Cys on serum IL-6 and TNF-α levels in mice with acute liver injury. ##P<0.01 vs Control group; **P<0.01 vs LPS/D-GaIN group.

Fig.4 Pathological changes in the liver of the mice in different groups (HE staining, original magnification: ×200). A: Control group; B: LPS/D-GalN group; C: LPS/D-GalN+rSj-Cys group; D: rSj-Cys group.

Fig.5 　Effect of rSj-Cys on CD68 expression in liver tissue of mice with acuteliver injury (Immunohistochemistry, ×400). A: Blank control group; B: LPS/DGalNgroup; C: LPS/D-GalN+rSj-Cys group; D: rSj-Cys control group; ##P<0.01 vs normal group; **P<0.01 vsLPS/D-GaIN group.

Fig.6 Effect of rSj-Cys on Bax and Bcl-2 expression in the liver tissue of mice with acute liver injury. #P<0.05 vs normal group; ##P<0.01 vs normal group; *P<0.05 vs normal group; **P<0.01 vs model group.

Fig.7 　Effect of rSj-Cys on hepatocyte apoptosis in the livertissue of mice with acute liver injury detected by TUNELstaining (×200). Normal cell nuclei are stained blue andapoptotic cell nuclei are stained red. ##P<0.01 vs Controlgroup; **P<0.01 vs LPS/D-GaIN group.

Fig.8 Effect of rSj-Cys on expressions of endoplasmic reticulum stress-related signaling pathway proteins in mice with acute liver injury. ##P<0.01 vs Control group; *P<0.05 vs, **P<0.01 vs LPS/D-GaIN group.

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