Roles of nuclear factor-κB/myeloid differentiation factor 88 in the liver fibrosis of cystic echinococcosis patients

doi:10.12140/j.issn.1000-7423.2021.06.008

Abstract

Abstract:

Objective To investigate roles of nuclear factor-κB/myeloid differentiation factor 88 (NF-κB/MyD88) in the liver fibrosis of patients with cystic echinococcosis (CE). Methods Liver samples were collected from the liver lesion tissue surgically resected from CE patients, by taking the tissue proximal to the lesion (0.5 cm to the outer cyst wall) and distant normal tissue (2 cm to outer cyst wall). Pathological changes of liver samples were observed by HE staining. Collagen deposition was observed with Masson staining. Expression of hepatic stellate cell activation marker α-smooth muscle actin (α-SMA), NF-κB p65 and MyD88 in the liver tissues of CE patients was detected by immunohistochemical staining and qRT-PCR analysis. Pearson correlation coefficient method was used to analyze the correlation of the expression level of NF-κB p65 and MyD88 with the positive area of collagen deposition, and expression level of α-SMA. Results Forty liver lesion samples resected from CE patients were collected, among them there were eight CE1, ten CE2, seven CE3, nine CE4 and six CE5. Of the patients receiving surgery, 24 were males and 16 females, aged from 20 to 45 years with the median of (38.2 ± 12.9) year. HE staining showed that inflammatory microenvironment was formed between CE lesion and proximal liver tissue, presenting massive lymphocyte infiltration, accompanied by increase of granulocyte to different degree. Masson staining showed that collagen was mainly deposited in the inflammatory cell band in the tissue proximal to lesion, around portal areas, biliary ducts and blood vessels. The positive area of collagen deposition in proximal-lesion tissue (20.29 ± 3.96)% was higher than that (2.87 ± 1.74)% in distant normal liver tissue (t = 13.640, P < 0.01). Immunohistochemical staining demonstrated that α-SMA, NF-κB p65 and MyD88 positive cells mainly highly expressed in the inflammatory cell band around proximal-lesion tissue, which was (13.47 ± 3.47)%, (7.30 ± 1.40)% and (7.47 ± 1.86)%, respectively. However, their expression in distant normal liver tissue was quite low, which was (3.43 ± 0.56)%, (1.08 ± 0.29)%, (0.36 ± 0.05)%, respectively, showing significant difference of expression in proximal and distant location(t = 5.682, 18.530, 5.087, P < 0.01). qRT-PCR analysis indicated that relative expression amount of α-SMA, NF-κB p65, MyD88 mRNA in proximal-lesion samples was 5.05 ± 0.42, 3.71 ± 0.33, 7.11 ± 0.50, respectively, which was significantly higher than those in the distant normal tissues (1.07 ± 0.01, 1.02 ± 0.03, 1.07 ± 0.02, respectively) (t = 39.750, 34.130, 50.960, P < 0.01). The expression of NF-κB p65 and MyD88 was significantly correlated with the and positive area of collagen deposition and α-SMA expression (r = 0.98, 0.97, 0.98, P < 0.01). Conclusion NF-κB p65/MyD88 expression in the proximal-lesion liver tissue of hepatic CE patients was significantly higher than that in the distant normal liver tissue, which was positively correlated with the degree of liver fibrosis.

Key words: Cystic echinococcosis, NF-κB, MyD88, Liver fibrosis

CLC Number:

R532.32

MA Wen-mei, SANG Wei, AIMAITI Ya-sen, ZUO Li-ke, FU Li, MIAO Na. Roles of nuclear factor-κB/myeloid differentiation factor 88 in the liver fibrosis of cystic echinococcosis patients[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2021, 39(6): 779-783.

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References 19

[1]	Hao W, Lucine V, Tuerhongjiang T, et al. Echinococcosis in the 21st century[J]. Clin Microbiol Rev, 2019, 32(2): e00075-93.
[2]	Budke CM, Carabin H, Patrick CN, et al. A systematic review of the literature on cystic echinococcosis frequency worldwide and its associated clinical manifestations[J]. Am J Trop Med Hyg, 2013, 88(6): 1011-1027. doi: 10.4269/ajtmh.12-0692
[3]	Craig PS, Larrieu E. Control of cystic echinococcosis/hydatidosis: 1863—2002[J]. Adv Parasitol, 2006, 61: 443-508.
[4]	Labsi M, Soufli I, Khelifi L, et al. In vivo treatment with IL-17A attenuates hydatid cyst growth and liver fibrogenesis in an experimental model of echinococcosis[J]. Acta Trop, 2018, 181: 6-10. doi: 10.1016/j.actatropica.2018.01.014
[5]	Hong KK, Gwak MJ, Song J, et al. NF-κB pathway activation and PTEN downregulation in psoriasis[J]. Br J Dermatol, 2016, 174(2): 433-435. doi: 10.1111/bjd.14106 pmid: 26302365
[6]	Tsuruta D. NF-kappaB links keratinocytes and lymphocytes in the pathogenesis of psoriasis[J]. Recent Pat Inflamm Allergy Drug Discov, 2009, 3(1): 40-48. doi: 10.2174/187221309787158399
[7]	Ji R, Liang RW, Guan ZY, et al. The role of TLR4/NF-κB signaling pathway in Cryptosporidim parvum infection[J]. Chin J Parasitol Parasit Dis, 2018, 36(4): 361-365. (in Chinese)
	(汲蕊, 梁瑞文, 管志玉, 等. TLR4/NF-κB信号通路在微小隐孢子虫感染中的作用机制[J]. 中国寄生虫学与寄生虫病杂志, 2018, 36(4): 361-365.)
[8]	Wang S, Zhang Z, Wang Y, et al. Toxoplasma gondii excretory/secretory antigens (TgESAs) suppress pro-inflammatory cytokine secretion by inhibiting TLR-induced NF-κB activation in LPS-stimulated murine macrophages[J]. Oncotarget, 2017, 8(51): 88351-88359. doi: 10.18632/oncotarget.v8i51
[9]	Zhang C, Shao Y, Yang S, et al. T-cell tolerance and exhaustion in the clearance of Echinococcus multilocularis: role of inoculum size in a quantitative hepatic experimental model[J]. Sci Rep, 2017, 7(1): 11153. doi: 10.1038/s41598-017-11703-1
[10]	Lopez-Luis BA, Valdivia-Cayoja AR, Belaunzaran-Zamudio PF, et al. An immunocompromised patient & multiorgan cystic echinococcosis[J]. QJM-INT J MED, 2019, 112(3): 215-217. doi: 10.1093/qjmed/hcy306
[11]	Joyce JA, Fearon DT. T cell exclusion, immune privilege, and the tumor microenvironment[J]. Science, 2015, 348(6230): 74-80. doi: 10.1126/science.aaa6204
[12]	Reynaert H, Thompson MG, Thomas T, et al. Hepatic stellate cells: roles in mivrovirulation and pathophysiology of portal hypertension[J]. Gat, 2002, 50: 571-581.
[13]	Dietrich GG, Gotze O, Geier A. Molecular changes in hepatic metabolism and transport in cirrhosis and their functional importance[J]. World J Gastroentrol, 2016, 22(1): 72-88.
[14]	Cai JY, Shao YX, Wang K, et al. Paeoniflorin inhibits activation of Raw 264.7 macrophages induced by high glucose via JAK2/STAT3 signaling pathway[J]. Chin Pharmacol Bull, 2019, 35(1): 56-62. (in Chinese)
	(蔡建月, 邵云侠, 王坤, 等. 芍药苷通过JAK2/STAT3信号通路抑制高糖诱导的RAW264.7巨噬细胞激活[J]. 中国药理学通报, 2019, 35(1): 56-62.)
[15]	Zhong J, Wang H, Chen W, et al. Ubiquitylation of MFHAS1 by the ubiquitin ligase praja2 promotes M1 macrophage polarization by activating JNK and p38 pathways[J]. Cell Death Dis, 2017, 8(5): e2763. doi: 10.1038/cddis.2017.102
[16]	Nakano T, Fukuda D, Koga J, et al. Delta-Like ligand 4-Notch signaling in macrophage activation[J]. Arterioscler Thromb Vasc Biol, 2016, 36(10): 2038-2047. doi: 10.1161/ATVBAHA.116.306926
[17]	Czimmerer Z, Daniel B, Horvath A, et al. The transcription factor STAT6 mediates direct repression of inflammatory enhancers and limits activation of alternatively polarized macrophages[J]. Immunity, 2018, 48(1): 75-90. doi: S1074-7613(17)30568-X pmid: 29343442
[18]	Sarah T, Dalila M, Zine-Charaf AT, et al. Potential role of NF-κB pathway in the immuno-inflammatory responses during human cystic echinococcosis[J]. Acta Trop, 2020, 203(8): 105306. doi: 10.1016/j.actatropica.2019.105306
[19]	Caamano J, Hunter CA. NF-κB family of transcription factors: central regulators of innate and adaptive immune functions[J]. Clin Microbiol Rev, 2002, 15(3): 414-429. doi: 10.1128/CMR.15.3.414-429.2002

基因名称Gene name	引物序列（5′→3′） Primer sequence (5′→3′)
α-平滑肌肌动蛋白 ɑ-SMA	F: AGGCACCCCTGAACCCCAA
	R: CAGCACCGCCTGGATAGCC
核因子-κB p65 NF-κB p65	F: AACAGCAGATGGCCCATACCT
	R: ACGCTGAGGTCCATCTCCTTG
髓样分化分子88 MyD88	F: GGCTGCTCTCAACATGCGA
	R: CTGTGTCCGCACGTTCAAGA
甘油醛-3-磷酸脱氢酶 GAPDH	F: CTGCTCCTCCTGTTCGACAGT
	R: CCGTTGACTCCGACCTTCAC