Tianjin Medical Journal ›› 2022, Vol. 50 ›› Issue (1): 5-10.doi: 10.11958/20211111

• Monograph-Biliary Atresia and Liver Transplantation • Previous Articles     Next Articles

Study on the mechanism of EGF-ERK1/2-IL-8 pathway in cholangitis after Kasai procedure in biliary atresia

ZHENG Qipeng1, 2, LI Mengdi1, 2, ZHANG Cong1, 2, CHEN Lingzhi1, 2, ZHAO Yilin1, 2, YANG Fang1, 2, LIU Gengxin1, 2, ZHAN Jianghua2△#br#   

  1. 1 Graduate School, Tianjin Medical University, Tianjin 300070, China; 2 Department of General Surgery, 
    Tianjin Children's Hospital
  • Received:2021-05-12 Revised:2021-08-26 Published:2022-01-15 Online:2022-01-19
  • Contact: zhanjianghua E-mail:zhanjianghuatj@163.com

Abstract:

Abstract: Objective To investigate the expression level of interleukin-8 (IL-8) in children with biliary atresia (BA) and its relationship with cholangitis after Kasai procedure, and to explore the upstream signaling pathway regulating IL-8. Methods (1) A total of 33 children with BA (18 cases with cholangitis and 15 cases without cholangitis after Kasai procedure), 6 children with biliary dilatation and 36 healthy children were included in this study. Serum samples were collected from children with BA and healthy infants. The serum expression levels of IL-8 and epidermal growth factor (EGF) were determined by enzyme-linked immunosorbent assay (ELISA). The activation of extracellular signal-regulated kinase 1/2 (ERK1/2) was detected by immunohistochemical staining in liver tissue samples from 6 children with BA and 6 children with biliary dilatation. (2) Human normal hepatocytes L-02 were cultured in vitro, and treated with phosphate buffer (PBS) and 25, 50 and 100 μg/L EGF for 24 h. The level of IL-8 was detected by quantitative polymerase chain reaction (qPCR). Cell lines HIBEpic, L-02 and LX-2 were treated with PBS and 100 μg/L EGF for 24 h, and the levels of IL-8 were detected by qPCR. L-02 cells were treated with 100 μg/L EGF and/or 5 mmol/L ERK pathway inhibitor KO-947, and the activation of ERK1/2 and the expression level of IL-8 were detected. Results (1) The serum level of IL-8 was significantly higher in the BA group than that in the normal control group (P<0.01), and the serum level of IL-8 was significantly higher in the cholangitis group than that in the non-cholangitis group (P<0.01). Serum EGF was positively correlated with IL-8 in the BA group (rs=0.422, P<0.01). Immunohistochemical staining showed that p-ERK1/2 was highly expressed in liver tissues of BA group. (2) The IL-8 expression in L-02 cells increased in a concentration-dependent manner after PBS and 25, 50, 100 μg/L EGF intervention. There were no significant differences in the expression levels of IL-8 in HIBEpic and LX-2 cells after 100 μg/L EGF treatment (P>0.05), but the expression of L-8 was increased in L-02 cells (P<0.01). EGF at 100 μg/L also induced the expression of p-ERK1/2 in L-02 cells, and KO-947 could reverse the changes. Conclusion The serum IL-8 expression is significantly increased in children with BA before surgery, which is closely related to the postoperative cholangitis of Kasai. EGF could promote the expression of IL-8 in hepatocytes through ERK1/2 signaling pathway. 

Key words: biliary atresia, cholangitis, interleukin-8, epidermal growth factor, extracellular signal-regulated MAP kinases

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