CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES >

2023 , Vol. 41 >Issue 6: 760 - 765

DOI: https://doi.org/10.12140/j.issn.1000-7423.2023.06.015

SHORT COMMUNICATIONS

Effect of Clonorchis sinensis infection on hepatic fibrosis and immune regulation in mice

  • ZHAO Lei ,
  • LI Jia ,
  • MO Gang ,
  • LI Chun ,
  • HUANG Guoyang ,
  • PENG Xiaohong
Expand
  • 1 Guangxi University Key Laboratory of Pathogenic Biology, Guilin 541004, China
    2 Guangxi Key Laboratory of Molecular Medicine in Liver Injury and Repair, the Affiliated Hospital of Guilin Medical University, Guilin 541004, China

Received date: 2023-06-21

  Revised date: 2023-07-18

  Online published: 2023-12-26

Supported by

National Science Foundation of China(82060376);Guangxi Province Postgraduate Co-training Base for Cooperative Innovation in Basic Medicine (Gui Xue Wei [2020]7)

Fold

Abstract

In order to further understand the liver fibrosis and immune regulatory changes in mice infected with Clonorchis sinensis, BALB/c mice were randomly divided into 2 groups with 30 mice in each group. The infected group were administered 200 μl normal saline with 150 C. sinensis metacercariae through oral gavage and the control group received 200 μl normal saline. Six mice were randomly selected for dissection at 1, 2, 4, 8 and 16 weeks. The paraffin sections of the liver tissue were stained with hematoxylin-eosin (HE) and Masson to observe the progression of fibrosis. The relative transcription level and actin alpha 2 (ACTA2) expression in the liver were detected by qRT-PCR and western blotting separately. The spleen tissue were weighed for calculating the spleen index (spleen weight per 10 g weight, mg/10 g). The flow cytometry was performed to detect the dynamical changes of CD4+ T cells, helper T1 (Th1) cells, Th2 cells, Th17 cells and regulatory T (Treg) cells in spleen. The secretion levels of tumor necrosis factor (TNF), interleukin-6 (IL-6), interferon-γ (IFN-γ), IL-2, IL-4, IL-10, and IL-17A were quantified by cytometric bead array. The data was analyzed by GraphPad Prism 9.0. Two-way ANOVA was used for groups comparison and t test was used for pairwise comparison. The results showed that the mice weight decreased and spleen index increased after infected with C. sinensis and the inflammatory lesions were visible in liver tissue. HE staining showed that C. sinensis larvae and inflammatory cells could be found around the hepatobiliary duct at week 1 after infection. Masson staining revealed that the relative area of collagen fiber deposition was higher than the control group after infection (F = 20.190, P < 0.05). The qRT-PCR results showed that the hepatic ACTA2 relative transcription level of the infected group were higher than the control group at week 2, 4 and 16 after infection (t = 2.042, 2.475, 1.634; all P < 0.01) but lower than the control group at week 8 (t = 2.758, P < 0.05). Western blotting results showed that the hepatic ACTA2 protein relative expression level of the infected group were higher than the control group (F = 3.225, P < 0.01). The flow cytometry results showed that the percentage of splenic CD4+ T cells decreased to (13.4 ± 1.8)% at week 2 after infection and increased to (22.4 ± 1.5)% at week 16. The percentage of Th1 cells increased to (16.9 ± 5.3)% at week 2 and decreased to (3.9 ± 2.6)% at week 16. The percentage of Th2 cells decreased to (2.3 ± 0.6)% at week 4 and increased gradually after week 4. The percentage of Th17 cells increased to (5.3 ± 3.4)% at week 8 and decreased to (2.4 ± 1.4)% at week 16. The percentage of Treg cells decreased to (7.3 ± 1.5)% at week 4 and increased to (13.9 ± 1.2)% at week 16. The serum TNF content of infected group mice increased to (35.16 ± 11.28) pg/ml at week 1 after infection and decreased to (8.98 ± 1.66) pg/ml at week 8. The serum IL-6 content was higher than the control group after infection (t = 2.095, P < 0.05). The serum IL-2 content decreased to (0.09 ± 0.18) pg/ml at week 8 and increased to (3.81 ± 2.79) pg/ml at week 16 after infection. The content of serum IFN-γ, IL-4, IL-10 and IL-17A increased gradually after infection (F = 8.726, 8.068, 6.795, 14.840; all P < 0.05). As a conclusion, the dynamic changes of CD4+ T cells and serum cytokines were closely related to the hepatic fibrosis induced by C. sinensis.

Key words： Clonorchis sinensis; Hepatic fibrosis; Splenic CD4+ T cells; Dynamic change

Cite this article

ZHAO Lei , LI Jia , MO Gang , LI Chun , HUANG Guoyang , PENG Xiaohong . Effect of Clonorchis sinensis infection on hepatic fibrosis and immune regulation in mice[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2023 , 41(6) : 760 -765 . DOI: 10.12140/j.issn.1000-7423.2023.06.015

References

[1] Qian MB, Utzinger J, Keiser J, et al. Clonorchiasis[J]. Lancet, 2016, 387(10020): 800-810.
[2] Zhao TT, Fang YY, Lai YS. Assessment of the burden of clonorchiasis and its temporal changes in China[J]. Chin J Schisto Control. 2021, 33(2): 162-168. (in Chinese)
  (赵婷婷, 方悦怡, 赖颖斯. 中国华支睾吸虫病疾病负担估算及其变化趋势分析[J]. 中国血吸虫病防治杂志, 2021, 33(2): 162-168.)
[3] Xin HL, Yang YC, Jiang ZH, et al. An investigation of human clonorchiasis prevalence in an endemic county in Guangxi Zhuang Autonomous Region, China, 2016[J]. Food Waterborne Parasitol, 2021, 22: e00109.
[4] Zeng XM, Jiang ZH, Shen JQ, et al. Detection and analysis of fecal egg and serum IgG antibody in 146 suspected cases of Clonorchis sinensis infection in Guangxi[J]. Chin J Parasitol Parasit Dis, 2019, 37(6): 730-732. (in Chinese)
  (曾雪梅, 蒋智华, 申继清, 等. 广西146例华支睾吸虫疑似感染者粪样虫卵和血清IgG抗体的检测分析[J]. 中国寄生虫学与寄生虫病杂志, 2019, 37(6): 730-732.)
[5] Huang GH, Zhang B, Lai HT, et al. Analysis on the prevalence of Clonorchis sinensis infection in Lingshan County, Guangxi from 2016 to 2019[J]. Chin J Parasitol Parasit Dis, 2022, 40(5): 673-676. (in Chinese)
  (黄光华, 张波, 赖海涛, 等. 2016—2019年广西灵山县人群华支睾吸虫感染情况分析[J]. 中国寄生虫学与寄生虫病杂志, 2022, 40(5): 673-676.)
[6] Bai XL, Wang N, Zhou J, et al. DX5+ NKT cells’ increase was correlated with liver damage in FVB mice not in BALB/c mice infected by Clonorchis sinensis[J]. Parasite Immunol, 2021, 43(1): e12796.
[7] Wang N, Bai X, Jin XM, et al. The dynamics of select cellular responses and cytokine expression profiles in mice infected with juvenile Clonorchis sinensis[J]. Acta Trop, 2021, 217: 105852.
[8] Hung KS, Lee TH, Chou WY, et al. Interleukin-10 gene therapy reverses thioacetamide-induced liver fibrosis in mice[J]. Biochem Biophys Res Commun, 2005, 336(1): 324-331.
[9] Zhou WC, Zhang QB, Qiao L. Pathogenesis of liver cirrhosis[J]. World J Gastroenterol, 2014, 20(23): 7312-7324.
[10] Zhang MJ, Zhang S. T cells in fibrosis and fibrotic diseases[J]. Front Immunol, 2020, 11: 1142.
[11] Glimcher LH, Murphy KM. Lineage commitment in the immune system: the T helper lymphocyte grows up[J]. Genes Dev, 2000, 14(14): 1693-1711.
[12] Elson CO, Cong Y, Brandwein S, et al. Experimental models to study molecular mechanisms underlying intestinal inflammation[J]. Ann N Y Acad Sci, 1998(859): 85-95.
[13] Lee GR. The balance of Th17 versus Treg cells in autoimmunity[J]. Int J Mol Sci, 2018, 19(3): 730.
[14] G?schl L, Scheinecker C, Bonelli M. Treg cells in autoimmunity: from identification to Treg-based therapies[J]. Semin Immunopathol, 2019, 41(3): 301-314.
[15] Hanley CJ, Waise S, Ellis MJ, et al. Single-cell analysis reveals prognostic fibroblast subpopulations linked to molecular and immunological subtypes of lung cancer[J]. Nat Commun, 2023, 14(1): 387.
[16] Zhao L, Mo G, Li J, et al. Establishment of a model of hepatic bile duct fibrosis in BALB/c mice infected with Clonorchis sinensis[J]. J Pathog Biol, 2022, 17(10): 1160-1163. (in Chinese)
  (赵磊, 莫刚, 李佳, 等. 华支睾吸虫感染BALB/c小鼠肝胆管纤维化模型的建立[J]. 中国病原生物学杂志, 2022, 17(10): 1160-1163.)
[17] Zhao L, Shi MC, Zhou LN, et al. Clonorchis sinensis adult-derived proteins elicit Th2 immune responses by regulating dendritic cells via mannose receptor[J]. PLoS Negl Trop Dis, 2018, 12(3): e0006251.
[18] Taylor AE, Carey AN, Kudira R, et al. Interleukin 2 promotes hepatic regulatory T cell responses and protects from biliary fibrosis in murine sclerosing cholangitis[J]. Hepatology, 2018, 68(5): 1905-1921.
[19] Yan C, Zhang BB, Hua H, et al. The dynamics of Treg/Th17 and the imbalance of Treg/Th17 in Clonorchis sinensis-infected mice[J]. PLoS One, 2015, 10(11): e0143217.
[20] Dirchwolf M, Podhorzer A, Marino M, et al. Immune dysfunction in cirrhosis: distinct cytokines phenotypes according to cirrhosis severity[J]. Cytokine, 2016, 77: 14-25.
[21] Rey I, Effendi-Ys R. Association between serum IL-6, IL-10, IL-12, and IL-23 levels and severity of liver cirrhosis[J]. Med Arch, 2021, 75(3): 199-203.
[22] Velavan TP, Ojurongbe O. Regulatory T cells and parasites[J]. J Biomed Biotechnol, 2011, 2011: 520940.
[23] Ma X, Hua J, Mohamood AR, et al. A high-fat diet and regulatory T cells influence susceptibility to endotoxin-induced liver injury[J]. Hepatology, 2007, 46(5): 1519-1529.
Options
Outlines

/

〈 〉