Echinococcus granulosus cyst fluid promotes activation and migration of hepatic stellate cells

doi:10.12140/j.issn.1000-7423.2025.01.011

Abstract

Abstract:

Objective To explore the effects and mechanisms of Echinococcus granulosus cyst fluid (CF) on the activation and migration of hepatic stellate cells (HSC). Methods Liver lesion tissues of cystic echinococcosis (CE) patients were obtained, and the fibrosis of liver tissues was observed using hematoxylin and eosin (HE) staining and Masson staining. HSCs were cultured in vitro for 48 and 72 hours in culture medium without CF and with 10%, 20% and 40% CF, respectively. Western blotting was used to detect the relative protein expression levels of α-mooth muscle actin (α-SMA), collagen type I alpha 1 (Col1a1), phosphatidylinositol 3 kinase (PI3K), phosphorylated PI3K (p-PI3K), protein kinase B (Akt) and phosphorylated Akt (p-Akt). Real-time quantitative PCR (qPCR) was used to detect the relative mRNA transcription levels of α-SMA, Col1a1 and Col3a1. Scratch assay and Transwell assays were conducted to assess the effects of CF on HSC migration. Results HE staining showed that the outer capsule of E. granulosus in the liver lesion tissues of CE patients was transparent and homogeneous with fibrous strips, and Masson staining showed a large amount of collagen fiber deposition in the outer capsule. Western blotting results showed that after 72 hours of intervention, the relative protein expression levels of α-SMA in the control group, 10% CF group, 20% CF group and 40% CF group were 0.359 ± 0.043, 0.641 ± 0.088, 0.900 ± 0.084 and 1.111 ± 0.027, respectively. The 10% CF group, 20% CF group, and 40% CF group had higher levels than the control group (t = 5.236, 10.050, 13.980; all P < 0.05). The relative expression levels of Col1a1 in each group were 0.392 ± 0.057, 0.546 ± 0.022, 0.854 ± 0.034 and 1.127 ± 0.057, respectively. The 10% CF group, 20% CF group and 40% CF group had higher levels than the control group (t = 4.228, 12.630, 20.090; all P < 0.05). qPCR results showed that after 72 hours of intervention, the relative mRNA transcription levels of α-SMA in the control group, 10% CF group, 20% CF group and 40% CF group were 1.000 ± 0.093, 1.437 ± 0.106, 1.453 ± 0.105 and 1.697 ± 0.154, respectively. The 10% CF group, 20% CF group and 40% CF group had higher levels than the control group (t = 4.604, 4.769, 7.339; all P < 0.05). The relative mRNA transcription levels of Col1a1 in each group were 0.856 ± 0.042, 1.067 ± 0.049, 1.283 ± 0.128 and 1.582 ± 0.046, respectively. The 20% CF group and 40% CF group had higher levels than the control group (t = 6.932, 11.790; both P < 0.05). The relative mRNA transcription levels of Col3a1 in each group were 0.611 ± 0.054, 0.908 ± 0.041, 1.000 ± 0.045 and 1.239 ± 0.101, respectively. The 10% CF group, 20% CF group and 40% CF group had higher levels than the control group (t = 5.599, 7.346, 11.850; all P < 0.05). Western blotting results showed that the relative protein expression levels of p-PI3K/PI3K in the control group, 10% CF group, 20% CF group and 40% CF group were 0.346 ± 0.050, 0.716 ± 0.054, 0.941 ± 0.114 and 1.276 ± 0.004, respectively. The 10% CF group, 20% CF group, and 40% CF group had higher levels than the control group (t = 6.669, 10.740, 16.780; all P < 0.01). The relative protein expression levels of p-Akt/Akt in each group were 0.524 ± 0.111, 0.815 ± 0.019, 1.043 ± 0.052 and 1.333 ± 0.054, respectively. The 10% CF group, 20% CF group and 40% CF group had higher levels than the control group (t = 5.268, 9.413, 14.670; all P < 0.01). Scratch assay results showed that after 24 hours of intervention, the cell migration rates in the 10% CF group, 20% CF group and 40% CF group were (73.6 ± 2.1)%, (88.2 ± 2.1)% and (96.5 ± 1.2)%, respectively, all higher than (55.1 ± 2.9)% in the control group (t = 10.510, 18.820, 23.540; all P < 0.05). Transwell assay results showed that after 24 hours of intervention, the average number of cells per field in the control group, 10% CF group, 20% CF group and 40% CF group were (62.333 ± 8.145), (82.333 ± 8.505), (132.333 ± 17.620) and (164.333 ± 11.060), respectively. The 20% CF group and 40% CF group had higher numbers than the control group (t = 7.173, 10.450; both P < 0.01). Conclusion E. granulosus CF could promote the activation of HSC and enhance their migration ability through the PI3K/Akt pathway.

Key words: Echinococcus granulosus, Hepatic stellate cells, Cyst fluid, Fibrosis

CLC Number:

R532.32

CUI Jie, WU Xiya, LIN Zhiyi, CAI Kefeng, WU Zhengzhan, XIONG Zitong, HUANG Yanxin, FANG Fuzhong, XIN Zirui, ZHANG Hongwei. Echinococcus granulosus cyst fluid promotes activation and migration of hepatic stellate cells[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2025, 43(1): 69-75.

Figures/Tables 7

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基因 Gene	引物序列（5′→3′） Primer sequence (5′→3′)
α-SMA	F：CTATGAGGGCTATGCCTTGCC
	R：GCTCAGCAGTAGTAACGAAGGA
Col3a1	F：TTGAAGGAGGATGTTCCCATCT
	R：ACAGACACATATTTGGCATGGTT
Col1a1	F：GTGCGATGACGTGATCTGTGA
	R：CGGTGGTTTCTTGGTCGGT
β-actin	F：AGTGTGACGTTGACATCCGTA
	R：CCAGAGCAGTAATCTCCTTCT