Screening of marker of hepatic stellate cells apoptosis in schistosomiasis japonica-induced liver fibrosis

doi:10.12140/j.issn.1000-7423.2025.04.005

Abstract

Abstract:

Objective To screen molecular markers suitable for labeling host hepatic stellate cell (HSC) apoptosis in schistosomiasis japonica-induced liver fibrosis (SSLF). Methods Amino acid sequences of apoptosis-related proteins from Mus musculus and Schistosoma japonicum were obtained through literature review and database retrieval, and homology analysis of these proteins was performed using the Protein BLAST tool on the NCBI website. Liver tissues were collected from mice infected with S. japonicum 4, 8 and 12 weeks post-infection, and uninfected mice served as controls. Liver tissues were sectioned, and the expression of HSC marker Desmin and apoptosis-related molecules was detected using immunofluorescence staining. The dynamics of mouse HSC apoptosis were analyzed at different time points post-infection to identify molecular markers indicative of host cell apoptosis in SSLF. In addition, sequence homology and phylogenetic analysis of apoptosis-related proteins sequences from different adaptive hosts of S. japonicum were performed using the software Jalview and MEGA11. All data were analyzed with one-way analysis of variance (ANOVA) using the software GraphPad Prism 8. Results Sixteen S. japonicum apoptosis-related proteins and eighteen M. musculus apoptosis-related proteins were identified through literature review and database retrieval. Sequence similarity analysis showed 20% to 80% homology between sequences of M. musculus and S. japonicum apoptosis-related proteins, and no homologous proteins of M. musculus Fas or Fas ligand (FasL) were found in S. japonicum. Immunofluorescence staining showed remarkable distribution of apoptosis-related proteins (green fluorescence) Cleaved caspase 3, B cell-lymphoma-2 associated death promoter (Bad), and FasL surrounding egg granulomas in mice 8 weeks post-infection. Notably, Cleaved caspase 3 and Bad bound specifically to both mouse liver tissues and schistosome eggs, while FasL exclusively labeled host liver tissues, and was co-localized significantly with Desmin (red fluorescence). More importantly, the intensity of FasL and Desmin co-localization increased over time points post-infection. The proportion of FasL⁺Desmin⁺ cells (apoptotic HSCs) in Desmin⁺ cells (HSCs) was comparable between infected (0) and uninfected mice (0) 4 weeks post-infection (P > 0.05), and the proportions were 35.36% and 89.12% in infected mice 8 and 12 weeks post-infection, which were both significantly higher than in the uninfected group (t = 13.38, 28.77; both P < 0.01). Sequence alignment analysis showed that the homology of M. musculus FasL sequence was 91.04% with Rattus norvegicus, 80.57% with Microtus fortis, 75.09% with Homo sapiens, 76.24% with Sus scrofa, 76.34% with Ovis aries, 77.99% with Bubalus bubalis, 75.27% with Equus caballus, 74.48% with Canis lupus familiaris, and 77.86% with Felis catus. Phylogenetic analysis indicated that M. musculus FasL sequence was more closely related to R. norvegicus than to M. fortis, H. sapiens, S. scrofa, O. aries, B. bubalis, E. caballus, C. lupus familiaris, and F. catus. Conclusion FasL may be feasible tolabel host HSCs apoptosis in schistosomiasis japonica-induced liver fibrosis.

Key words: Schistosomiasis japonica, Liver fibrosis, Hepatic stellate cell apoptosis, Fas ligand

CLC Number:

R532.21

DONG Bowen, ZHONG Haoran, ZHU Danlin, LI Hao, LU Ke, FU Zhiqiang, LIU Jinming, JIN Yamei. Screening of marker of hepatic stellate cells apoptosis in schistosomiasis japonica-induced liver fibrosis[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2025, 43(4): 475-481.

Figures/Tables 4

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日本血吸虫凋亡蛋白 S. japonicum-derived apoptotic molecules	蛋白登录号 Accession no.	小鼠凋亡蛋白 M. musculus-derived apoptotic molecules	蛋白登录号 Accession no.	序列相似性/% Similarity/%
SjAIF	TNN08053.1	AIF	NP_001277293.1	32.75
SjAPAF	KAH8853392.1	APAF	NP_001269876.1	23.42
SjATM	KAH8866387.1	ATM	AAH67212.1	33.75
SjBad	AAW26442	Bad	NP_001272382.1	28.12
SjBak	CAX70134.1	Bak	NP_031549.2	36.00
SjBax	KAH8866109.1	Bax	NP_001398925.1	29.31
SjBcl2	KAH8856581.1	Bcl2	NP_033871.2	34.43
SjBIRP	TNN15698.1	BIRC6	NP_031592.3	60.19
SjCaspase 3	KAH8863222.1	Caspase 3	NP_001271338.1	39.38
SjCaspase 7	TNN10203.1	Caspase 7	NP_031637.1	40.40
SjCaspase 8	TNN08539.1	Caspase 8	NP_001073595.1	42.57
SjCaspase 9	Sjp_0071380	Caspase 9	NP_001264861.1	31.92
SjCIAP	TNN04927.1	Anamorsin	NP_598902.1	30.65
SjCyc	AAP06143.1	Cyc	AEP27250.1	80.00
/	/	Fas	NP_001140180.1	0
/	/	FasL	NP_034307.1	0
SjIAP	AAV97787.1	IAP1	NP_031490.2	46.00
Sjp53	TNN06778.1	TP53	NP_001120705.1	35.40
SjTNFR	TNN08705.1	TNF-R	NP_035739.2	30.14