细胞外基质蛋白1在多房棘球蚴感染小鼠肝纤维化过程中的作用

doi:10.12140/j.issn.1000-7423.2021.03.002

中国寄生虫学与寄生虫病杂志 ›› 2021, Vol. 39 ›› Issue (3): 296-303.doi: 10.12140/j.issn.1000-7423.2021.03.002

细胞外基质蛋白1在多房棘球蚴感染小鼠肝纤维化过程中的作用

李文定¹(), 温浩¹, 侯娇¹, 王明坤², 李亮¹, 李静², 张传山¹^,², 孙兵³, 王慧¹^,²^,^*()

1 新疆医科大学第一附属医院,临床医学研究院,省部共建中亚高发病成因与防治国家重点实验室,乌鲁木齐 830054
2 新疆医科大学基础医学院,乌鲁木齐 830054
3 中国科学院分子细胞科学卓越创新中心,生物化学与细胞生物学研究所,中国科学院大学,上海 200031

收稿日期:2021-02-04 修回日期:2021-03-22 出版日期:2021-06-30 发布日期:2021-07-05
通讯作者: 王慧
作者简介:李文定（1992-）,男,硕士研究生,从事棘球蚴感染所致宿主肝纤维化研究。E-mail： wendingli8506@163.com
基金资助:
新疆维吾尔自治区重点实验室开放课题(2019D04021);国家自然科学基金(81860359);省部共建中亚高发病成因与防治国家重点实验室开放课题项目(SKL-HIDCA-2020-5)

Role of extracellular matrix protein 1 in the liver fibrosis induced by Echinococcus multilocularis infection in mice

LI Wen-ding¹(), WEN Hao¹, HOU Jiao¹, WANG Ming-kun², LI liang¹, LI Jing², ZHANG Chuan-shan¹^,², SUN Bing³, WANG Hui¹^,²^,^*()

1 State Key Laboratory of Pathogenesis, Prevention and Treatment of High Incidence Diseases in Central Asia, Clinical Medicine Institute, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054,China
2 Basic Medical College, Xinjiang Medical University, Urumqi 830054, China
3 Chinese Academy of Sciences Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China

Received:2021-02-04 Revised:2021-03-22 Online:2021-06-30 Published:2021-07-05
Contact: WANG Hui
Supported by:
Xinjiang Uyghur Autonomous Region Key Laboratory Open Research Program Project(2019D04021);National Natural Science Foundation of China(81860359);State Key Laboratory of Pathogenesis, Prevention and Treatment of Central Asia High Incidence Diseases Fund(SKL-HIDCA-2020-5)

摘要/Abstract

摘要：

目的探讨多房棘球蚴不同感染时间小鼠肝组织细胞外基质蛋白1（extracellular matrix protein 1,ECM1）的表达水平以及与肝纤维化过程的相关性。方法将40只雌性 C57BL/6小鼠随机分为多房棘球蚴感染组和对照组（20只/组）,感染组经肝门静脉接种2 000个原头节/鼠,对照组注射等量生理盐水。分别于感染后第1、6、12和24周,两组各取5只小鼠采集肝组织,切片后HE染色观察多房棘球蚴感染后小鼠肝组织病理学变化;天狼星红染色以及α-SMA染色检测小鼠肝纤维化程度;免疫组织化学染色检测肝组织中ECM1的表达水平与分布。采用Pearson相关系数法分析ECM1表达水平与肝纤维化水平的相关性。结果 HE染色结果显示,多房棘球蚴感染后6周,小鼠肝组织内可见具有明显生发层结构的病灶形成,周围炎性细胞浸润明显且伴有纤维组织增生。天狼星红染色结果显示,感染组小鼠肝组织内病灶周围胶原沉积面积占比在感染后1、6、12和24周分别为（6.97 ± 0.07）%、（10.39 ± 0.02）%、（17.31 ± 1.78）%和（22.24 ± 1.07）%,均高于对照组（ P < 0.05）;α-SMA染色结果显示,感染组肝组织病灶周围α-SMA阳性染色面积占比在感染后1、6、12和24周分别为（5.31 ± 0.39）%、（9.97 ± 1.3）%、（16.16 ± 0.17）%和（19.01 ± 0.49）%,均高于对照组（ P < 0.05）;且随着多房棘球蚴感染小鼠时间的延长病灶周围肝纤维化程度不断加重。免疫组化检测结果显示,ECM1主要表达在病灶周围的炎性细胞带中,少量表达在肝窦;感染组ECM1阳性染色面积占比在感染后1、6、12和24周分别为（8.60 ± 0.44）%、（13.90 ± 0.57）%、（16.37 ± 0.77）%和（19.50 ± 0.50）%,均高于对照组（ P < 0.05）。Pearson相关性分析显示,ECM1表达水平与天狼星红阳性染色区域面积（ r = 0.900,P < 0.01）及α-SMA阳性染色区域面积占比（ r = 0.941,P < 0.01）均呈正相关。结论 ECM1在多房棘球蚴不同感染时间小鼠肝组织的表达均较对照组升高,并与肝纤维化程度呈正相关。

关键词: 多房棘球蚴, 感染, 肝纤维化, 细胞外基质蛋白1

Abstract:

Objective To investigate the role of extracellular matrix protein 1 (ECM1) in the liver fibrosis of mice infected with Echinococcus multilocularis (Em) by detecting the expression of ECM1 and the relationship with the liver fibrosis process. Methods Forty female C57BL/6 mice were randomly divided into the Em infection group (n = 20) and the control group (n = 20). For the Em infection group, 2 000 Em protoscoleces were collected under aseptic condition and inoculated through hepatic portal vein to establish the Em infection mouse model. Mice in the control group were injected with the same volume of saline in the same way. Liver tissues were collected at 1, 6, 12 and 24 weeks after Em infection. Histopathological changes were observed through hematoxylin eosin (HE) staining. The degree of liver fibrosis was evaluated through Sirius red staining and α-SMA staining. Immunohistochemical assay was performed to detect ECM1 expression in the mouse liver. The correlation between ECM1 expression and the degree of liver fibrosis was analyzed by Pearson post analysis. Results HE staining showed that lesions with obvious germinal layer structure were formed at 6 weeks after Em infection, showing obvious infiltration of inflammatory cells in surrounding tissues, accompanied by proliferation of fibrous tissue. Sirius red staining showed that the percentages of collagen deposition area in the mice liver tissue around the metacestodes lesion at week 1, 6 weeks, 12, and 24 after infection were (6.97 ± 0.07)%, (10.39 ± 0.02)%, (17.31 ± 1.78)%, and (22.24 ± 1.07)%, respectively, which were significantly higher than those in the control group ( P < 0.05). The α-SMA staining showed that the percentages of α-SMA-positive area around the lesion in the Em infection group were (5.31 ± 0.39)%, (9.97 ± 1.3)%, (16.16 ± 0.17)%, and (19.01 ± 0.49)%, respectively, at week 1, 6, 12, and 24 after infection, which were also significantly higher than that in the control group ( P < 0.05). These results indicated that the degree of liver fibrosis around the lesion gradually increased with the duration of Em infection. Immunohistochemical staining of ECM1 showed that ECM1 was mainly expressed in the inflammatory cell band around the lesion The percentages of ECM1-positive area in the Em infection group were (8.60 ± 0.44)%, (13.90 ± 0.57)%, (16.37 ± 0.77)%, and (19.50 ± 0.50)%, respectively, at week 1, 6, 12, and 24 after infection, which were significantly higher than those in the control group ( P < 0.05). Pearson correlation analysis showed that the expression level of ECM1 was positively correlated with the positive area with Sirius red staining ( r = 0.900, P < 0.01) and the percentage of α-SMA-positive area ( r = 0.941,P < 0.01). Conclusion ECM1 expression in the Em infected mouse liver at different time points post-infection was significantly higher than that in control group, and was positively correlated with the degree of liver fibrosis.

Key words: Echinococcus multiloculari, Infection, Liver fibrosis, Extracellular matrix protein 1

中图分类号:

R532.32

李文定, 温浩, 侯娇, 王明坤, 李亮, 李静, 张传山, 孙兵, 王慧. 细胞外基质蛋白1在多房棘球蚴感染小鼠肝纤维化过程中的作用[J]. 中国寄生虫学与寄生虫病杂志, 2021, 39(3): 296-303.

LI Wen-ding, WEN Hao, HOU Jiao, WANG Ming-kun, LI liang, LI Jing, ZHANG Chuan-shan, SUN Bing, WANG Hui. Role of extracellular matrix protein 1 in the liver fibrosis induced by Echinococcus multilocularis infection in mice[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2021, 39(3): 296-303.

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参考文献 20

[1]	Wen H, Vuitton L, Tuxun T, et al. Echinococcosis: advances in the 21st century[J]. Clin Microbiol Rev, 2019,32(2):e00075-e00018.
[2]	Aji T, Dong JH, Shao YM, et al. Ex vivo liver resection and auto transplantation as alternative to allotransplantation for end-stage hepatic alveolar echinococcosis[J]. J Hepatol, 2018,69(5):1037-1046. doi: 10.1016/j.jhep.2018.07.006
[3]	Hou XL, Li LH, Li L, et al. Changes in subsets and functional exhaustion of CD4⁺ T cells in spleens of mice infected with Echinococcus multilocularis[J]. Chin J Parasitol Parasit Dis, 2020,38(5):611-618, 624. (in Chinese)
	( 侯昕伶, 李玲慧, 李亮, 等. 多房棘球蚴感染小鼠脾CD4⁺ T细胞亚群及其功能耗竭的变化[J]. 中国寄生虫学与寄生虫病杂志, 2020,38(5):611-618, 624.)
[4]	Wen H. Hydatidosis [M]. Beijing: People’s Medical Publishing House, 2015: 553-554. (in Chinese)
	( 温浩. 包虫病学[M]. 北京: 人民卫生出版社, 2015: 553-554.)
[5]	Zhang CS. Study on TIGIT and PD-1 mediated T cells exhaustion in hepatic alveolar echinococcosis[D]. Urumqi: Xinjiang Medical University, 2017: 79-80. (in Chinese)
	( 张传山. TIGIT和PD-1介导泡型包虫病患者T淋巴细胞免疫耗竭的研究[D]. 乌鲁木齐: 新疆医科大学, 2017: 79-80.)
[6]	Fan WG, Liu TH, Chen W, et al. ECM1 prevents activation of transforming growth factor β, hepatic stellate cells, and fibrogenesis in mice[J]. Gastroenterology, 2019,157(5):1352-1367. doi: 10.1053/j.gastro.2019.07.036
[7]	Chen H, Jia WD, Li JS. ECM1 promotes migration and invasion of hepatocellular carcinoma by inducing epithelial-mesenchymal transition[J]. World J Surg Oncol, 2016,14(1):195. doi: 10.1186/s12957-016-0952-z pmid: 27460906
[8]	Xiong GP. Study on the role and mechanism of ECM1 in cholangiocarcinoma migration[D]. Fuzhou: Fujian Medical University, 2012: 47-50. (in Chinese)
	( 熊共鹏. ECM1在胆管癌转移中的作用及机制研究[D]. 福州: 福建医科大学, 2012: 47-50.)
[9]	Wang H, Li J, Guo BP, et al. In vitro culture of Echinococcus multilocularis producing protoscoleces and mouse infection with the cultured vesicles[J]. Parasit Vectors, 2016,9(1):411. doi: 10.1186/s13071-016-1687-y
[10]	Zhang CS, Shao YM, Yang ST, 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
[11]	Fukasawa H, Yamamoto T, Togawa A, et al. Down-regulation of Smad7 expression by ubiquitin-dependent degradation contributes to renal fibrosis in obstructive nephropathy in mice[J]. Proc Natl Acad Sci USA, 2004,101(23):8687-8692. doi: 10.1073/pnas.0400035101
[12]	Wu H, Chen GY, Wang JY, et al. TIM-4 interference in Kupffer cells against CCL4-induced liver fibrosis by mediating Akt1/mitophagy signalling pathway[J]. Cell Prolif, 2020,53(1):e12731.
[13]	Martí-Rodrigo A, Alegre F, Moragrega ÁB, et al. Rilpivirine attenuates liver fibrosis through selective STAT1-mediated apoptosis in hepatic stellate cells[J]. Gut, 2020,69(5):920-932. doi: 10.1136/gutjnl-2019-318372 pmid: 31530714
[14]	Hardy SA, Mabotuwana NS, Murtha LA, et al. Novel role of extracellular matrix protein 1 (ECM1) in cardiac aging and myocardial infarction[J]. PLoS One, 2019,14(2):e0212230. doi: 10.1371/journal.pone.0212230
[15]	Han Z, Ni J, Smits P, et al. Extracellular matrix protein 1 (ECM1) has angiogenic properties and is expressed by breast tumor cells[J]. FASEB J, 2001,15(6):988-994. pmid: 11292659
[16]	Wang LP, Yu JY, Ni J, et al. Extracellular matrix protein 1 (ECM1) is over-expressed in malignant epithelial tumors[J]. Cancer Lett, 2003,200(1):57-67. doi: 10.1016/S0304-3835(03)00350-1
[17]	Liu LQ. Study on the function of extracellular matrix protein 1 in gastric cancer metastasis and its mechanism[D]. Jinan: Shandong University, 2019: 15-17. (in Chinese)
	( 刘林青. 细胞外基质蛋白1在胃癌转移中的作用及其机制研究[D]. 济南: 山东大学, 2019: 15-17.)
[18]	Niu L, Song XG, Wang N, et al. Tumor-derived exosomal proteins as diagnostic biomarkers in non-small cell lung cancer[J]. Cancer Sci, 2019,110(1):433-442. doi: 10.1111/cas.2019.110.issue-1
[19]	Jiang HJ, Gui XW, Guo LJ, et al. Expression and angiogenic effect of VEGFA/VEGFR2 in mice hepatic metacestode tissue of Echinococcus multilocularis[J]. Chin J Parasitol Parasit Dis, 2020,38(6):673-681. (in Chinese)
	( 姜慧娇, 桂显伟, 郭黎姣, 等. VEGFA/VEGFR2在小鼠肝多房棘球蚴组织血管生成中的表达及作用[J]. 中国寄生虫学与寄生虫病杂志, 2020,38(6):673-681.)
[20]	Wang JH, Cardoso R, Marreros N, et al. Foxp3⁺ T regulatory cells as a potential target for immunotherapy against primary infection with Echinococcus multilocularis eggs[J]. Infect Immun, 2018,86(10):e00542-18.

细胞外基质蛋白1在多房棘球蚴感染小鼠肝纤维化过程中的作用

Role of extracellular matrix protein 1 in the liver fibrosis induced by Echinococcus multilocularis infection in mice

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