Schistosoma japonicum cystatin has protective effects against

doi:10.12122/j.issn.1673-4254.2025.01.14

Abstract

Abstract:

Objective To evaluate the protective effect of Schistosoma japonicum cystatin (rSj-Cystatin) in a mouse mode of "two-hit" sepsis. Methods Sixty male C57BL/6 mice randomized equally into sham-operated group, protein group, "two-hit" modeling group, and protein intervention group. In the former two groups, the mice received an intraperitoneal injection of 100 μL PBS followed by exposure of the cecum and then by intraperitoneal injection of 100 μL PBS or 25 μg rSj-Cystatin 30 min later; In the latter two groups, 100 μL PBS containing LPS (5 mg/kg) was injected intraperitoneally 24 h before cecal ligation and puncture (CLP), and 100 μL PBS or 25 μg rSj-Cystatin were injected 30 min after CLP. At 12 h after rSj-Cystatin treatment, 6 mice from each group were sacrificed for detection of TNF-α, IL-6, IL-10, TGF-β, iNOS and Arg-1 in the serum, spleen, liver, lung and kidney tissues using ELISA, for examinations of liver, lung and kidney pathologies with HE staining, and for analysis of CD3⁺CD4⁺CD25⁺Foxp3⁺T cell percentage in the spleen using flow cytometry. The remaining mice were observed for general condition and 72-h survival. Results The 72-h survival rates in the 4 groups were 100%, 100%, 0% and 20%, respectively, showing significant differences between the latter two groups. The mouse models of "two-hit" sepsis exhibited obvious tissue pathologies and significant elevations of TNF-α and IL-6 in both the serum and tissue homogenate, which were significantly ameliorated by rSj-Cystatin treatment. Treatment with rSj-Cystatin also increased IL-10 and TGF-β levels and spleen CD3⁺CD4⁺CD25⁺Foxp3⁺T cell percentage. The septic mouse models also showed increased iNOS levels in all the detected tissues and a decreased Arg-1 level in the kidney, and these changes were obviously improved by rSj-Cystatin treatment. Conclusion rSj-Cystatin has a protective effect against "two-hit" sepsis in mice by regulating the inflammatory microenvironment.

Key words: Schistosoma japonicum cysteine protease inhibitors, lipopolysaccharide, cecal ligation and puncture, sepsis, immunoregulatory cytokines

Wenjuan DUO, Yixiang WANG, Jiaxing WANG, Xinlong XU, Linxian LI, Dongchen YANG, Qili SHEN, Lichun YANG, Xiaojing LIU, Qiwang JING, Liang CHU, Xiaodi YANG. Schistosoma japonicum cystatin has protective effects against "two-hit" sepsis in mice by regulating the inflammatory microenvironment[J]. Journal of Southern Medical University, 2025, 45(1): 110-117.

Figures/Tables 9

Tab. 1 Liver injury scoring criteria

Hepatocyte edema, congestion, inflammatory cell infiltration	Index
None	0
0%-25%	1
25%-50%	2
50%-70%	3
75%-100%	4

Tab. 2 Lung injury scoring criteria

Alveolar hemorrhage and septum thickening, inflammatory cell infiltration	Index
None	0
0%-25%	1
25%-50%	2
50%-70%	3
75%-100%	4

Tab. 3 Kidney injury scoring criteria

Injured renal tubules and shrunk glomerulu	Index
None	0
0%-25%	1
25%-50%	2
50%-70%	3
75%-100%	4

Fig.1 Survival of the mice within 72 h in each group (n=10; **P<0.01, ***P<0.001).

Fig.2 Histopathological staining and pathological injury scoring of mice in each group. ***P<0.001 vs Group C.

Fig.3 Proportion of CD3+CD4+CD25+Foxp3+ T cells in the spleen of mice in each experimental group. Representative three-dimension scatter diagrams of CD3+CD4+CD25+Foxp3+ T cells in the spleen of mice in each group. The percentage of CD3+CD4+CD25+Foxp3+ T cells in the splenocytes of mice in each group, *P<0.05, **P<0.01 vs Group C.

Fig.4 Changes of serum levels of cytokines in each group. **P<0.01, ***P<0.001 vs Group C.

Fig.5 Levels of inflammatory factors in the liver, lung, and kidney tissues of the mice in each group. *P<0.05, **P<0.01, ***P<0.001 vs Group C.

Fig.6 Expression of M1 and M2 type biomarkers iNOS and Arg-1 in the liver, lungs, and kidney tissues in each group. *P<0.05, **P<0.01, ***P<0.001 vs Group C.

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