Role of Cytokines on the Progression of Liver Fibrosis in Mice Infected with Echinococcus multilocularis

Figures & data

Table 1 Hepatic Infection of Echinococcus multilocularis Severity Score (Mouse)

Degree of Inflammation		Vesicles* (Number)	Abnormal Structures and Substances	Score
Hepatic Lobule	Portal Area and Surrounding
No inflammation	No inflammation	0	No obvious change	0
Degeneration, small amount spotted and focal necrosis	Mild inflammation (Inflammatory cells occupy less than 1/3 of the portal area)	1–3	Granuloma	1
Degeneration, large amount spotted and focal necrosis		4–6		2
Confluent necrosis or bridging necrosis	Moderate inflammation (Inflammatory cells occupy 1/3~2/3 of the portal area)	7–9	Laminated layer and germinal layer	3
Extensive bridging of necrosis and involved many hepatic lobules	Severe inflammation (Inflammatory cells occupy more than 2/3 of the portal area)	≥10	Brood capsule and protoscolex	4

Note: *The average number of vesicle counts was observed in at least 3 different fields under low power microscope (magnification, ×100).

Table 2 Primer Sequence

Gene (Mouse)	Primer (5ʹ→3ʹ)
iNOS	Forward:GCAGAGATTGGAGGCCTTGTG
	Reverse:GGGTTGTTGCTGAACTTCCAGTC
IL-10	Forward:ATAAGAGCAAGGCAGTGGAGC
	Reverse:GGCCTTGTAGACACCTTGGTC
TGF-β	Forward:AGCTGCGCTTGCAGAGATTA
	Reverse:GACAGCCACTCAGGCGTATC
α-SMA	Forward:GTCCCAGACATCAGGGAGTAA
	Reverse:TCGGATACTTCAGCGTCAGGA
Col1α1	Forward:TAAGGGTCCCCAATGGTGAGA
	Reverse:GGGTCCCTCGACTCCTACAT
MMP9	Forward:GGACCCGAAGCGGACATTG
	Reverse:CGTCGTCGAAATGGGCATCT
caspase 3	Forward:CTCGCTCTGGTACGGATGTG
	Reverse:TCCCATAAATGACCCCTTCATCA
GAPDH	Forward:TGGTGAAGCAGGCATCTGAG
	Reverse:TGAAGTCGCAGGAGACAACC

Abbreviations: iNOS, inducible nitric oxide synthase; IL-10, interleukin-10; TGF-β, transforming growth factor-β; α-SMA, α-smooth muscle actin; Col1α1, collagen type I alpha 1; MMP9, matrix metalloproteinase 9; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Figure 1 Pathological changes in the liver of mice infected with Echinococcus multilocularis (x200).

Note: Immune cell infiltration and fibrosis change are showed with arrows.

Figure 2 (A) Liver inflammatory severity scores Note, n=8 in each group, ♦Indicates the score of inflammatory severity in each mouse; (B) dynamic quantification of fibrosis area Note, ◊Indicates the average area percent in Masson staining of different group; (C) liver function in mice infected with Echinococcus multilocularis.

Note: *Compared with the control group, P < 0.05.

Abbreviations: ALT, alanine aminotransferase; AST, aspartate aminotransferase.

Figure 3 Dynamic changes of inflammatory cytokines in immunohistochemistry (x200).

Abbreviations: iNOS, inducible nitric oxide synthase; IL-10, interleukin-10; TGF-β, transforming growth factor-β.

Figure 4 (A and C) Dynamic quantification of cytokines’ expression in protein level; (B and D) Dynamic quantification of cytokines’ expression in mRNA level.

Notes: *Compared with the control group, P < 0.05; ^#Compared with the infection model of 2d group; ◊Compared with the infection model of 8d group.

Abbreviations: iNOS, inducible nitric oxide synthase; IL-10, interleukin-10; TGF-β, transforming growth factor-β; α-SMA, α-smooth muscle actin; Col1α1, collagen type I alpha 1; MMP9, matrix metalloproteinase 9.

Figure 5 Dynamic changes of fibrosis related cytokines and cell apoptosis in immunohistochemistry (x200).

Abbreviations: α-SMA, α-smooth muscle actin; Col1α1, collagen type I alpha 1; MMP9, matrix metalloproteinase 9.