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. 2018 Nov;22(11):5486-5493.
doi: 10.1111/jcmm.13819. Epub 2018 Aug 9.

rSjP40 suppresses hepatic stellate cell activation by promoting microRNA-155 expression and inhibiting STAT5 and FOXO3a expression

Dandan Zhu  1 , Chunzhao Yang  1 , Pei Shen  2 , Liuting Chen  1 , Jinling Chen  1 , Xiaolei Sun  1 , Lian Duan  3 , Li Zhang  1 , Jinhua Zhu  1 , Yinong Duan  1
Affiliations

rSjP40 suppresses hepatic stellate cell activation by promoting microRNA-155 expression and inhibiting STAT5 and FOXO3a expression

Dandan Zhu et al. J Cell Mol Med. 2018 Nov.

Abstract

Activation of hepatic stellate cells (HSCs) is the central event of the evolution of hepatic fibrosis. Schistosomiasis is one of the pathogenic factors which could induce hepatic fibrosis. Previous studies have shown that recombinant Schistosoma japonicum egg antigen P40 (rSjP40) can inhibit the activation and proliferation of HSCs. MicroRNA-155 is one of the multifunctional noncoding RNA, which is involved in a series of important biological processes including cell development, proliferation, differentiation and apoptosis. Here, we try to observe the role of microRNA-155 in rSjP40-inhibited HSC activation and explore its potential mechanisms. We found that microRNA-155 was raised in rSjP40-treated HSCs, and further studies have shown that rSjP40 enhanced microRNA-155 expression by inhibiting STAT5 transcription. Up-regulated microRNA-155 can down-regulate the expression of FOXO3a and then participate in rSjP40-inhibited expression of α-smooth muscle actin (α-SMA) and collagen I. Furthermore, we observed microRNA-155 inhibitor could partially restore the down-regulation of FOXO3a, α-SMA and collagen I expression in LX-2 cells induced by rSjP40. Therefore, our research provides further insight into the mechanism by which rSjP40 could inhibit HSC activation via miR-155.

Keywords: Schistosoma japonicum; hepatic fibrosis; hepatic stellate cells; microRNA-155; rSjP40.

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Figures

Figure 1
Figure 1
miR‐155 expression is up‐regulated in LX‐2 cells treated with rSjP40. The expression levels of miR‐155 in LX‐2 cells which were treated with rSjP40 at the concentration of 20 μg/mL for 24 or 48 h were detected by RT‐qPCR. *P < 0.05, compared to each untreated group. The data are presented as the mean ± SEM of at least three independent experiments
Figure 2
Figure 2
FOXO3a is the target gene of miR‐155. A, The wild and mutant binding sites of miR‐155 in FOXO3a 3′UTR sequences were shown. B, Expression levels of FOXO3a protein in LX‐2 cells transfected with miRNA‐155 mimic or inhibitor were determined using Western blot. *P < 0.05, compared to each NC group. C, Relative luciferase activities of wild FOXO3a 3′UTR in LX‐2 cells transfected with miRNA‐155 mimic or inhibitor were determined by dual‐luciferase reporter assay. *P < 0.05, compared to each NC group. D, Relative luciferase activities of mutant FOXO3a 3′UTR in LX‐2 cells transfected with miRNA‐155 mimic or inhibitor were determined by dual‐luciferase reporter assay. NS represents P > 0.05, compared to each NC group. All the data above are presented as the mean ± SEM of at least three independent experiments
Figure 3
Figure 3
miR‐155 inhibitor can partially block down‐regulation of FOXO3a, α‐SMA, collagen I expression by rSjP40. A, The expression of FOXO3a protein levels in LX‐2 cells which were treated with rSjP40 (20 μg/mL) after 24 h was detected by Western blot. *P < 0.05, compared to untreated group. B, The expression levels of FOXO3a protein in LX‐2 cells which were transfected with miRNA‐155 inhibitor or NC inhibitor and treated with or without rSjP40 were detected by Western blot. C, The expression levels of α‐SMA and collagen I protein in LX‐2 cells which were transfected with miRNA‐155 inhibitor or NC inhibitor and treated with or without rSjP40 were detected by Western blot. The data are presented as the mean ± SEM of at least three independent experiments. *P < 0.05, compared to untreated group. # P < 0.05, compared to NC inhibitor + rSjP40− group. $ P < 0.05, compared to miR‐155 inhibitor + rSjP40− group. & P < 0.05, compared to NC inhibitor + rSjP40 + group. NS represents P > 0.05 and shows that there is no significant statistical difference between the compared two groups
Figure 4
Figure 4
rSjP40‐mediated enhancement of miR‐155 promoter activity is related to STAT5 in LX‐2 cells. A, Luciferase activities of pGL3‐basic and pGL3‐promoter miR‐155 in LX‐2 cells were determined by dual‐luciferase reporter assay. *P < 0.05, compared to pGL3‐basic group. B, The effect of rSjP40 on the luciferase activities of pGL3‐basic or pGL3‐promoter miR‐155 in LX‐2 cells was determined by dual‐luciferase reporter assay. *P < 0.05, compared to each untreated group. C, STAT5 and P‐STAT5 protein expression levels in LX‐2 cells treated with rSjP40 at the concentration of 20 μg/mL were evaluated by Western blot. *P < 0.05, compared to each untreated group. D, Diagram of STAT5 binding sites in the miR‐155 promoter was shown. E, ChIP analysis was performed to confirm the binding of STAT5 to the miR‐155 promoter. M, DL2300 from SMOBIO. Lane 1 represents anti‐Histone H3 group for positive control group. Lanes 5, 9, 13 and 17 represent anti‐STAT5 group for target group. Lanes 2, 6, 10, 14 and 18 represent normal IgG group. Lanes 3, 7, 11, 15 and 19 represent input group. Lanes 4, 8, 12, 16 and 20 represent water group using ddH2O as the template for PCR. Primer RPL30 was used for the positive control group
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References

    1. Mitra A, Satelli A, Yan J, et al. IL‐30 (IL27p28) attenuates liver fibrosis through inducing NKG2D‐rae1 interaction between NKT and activated hepatic stellate cells in mice. Hepatology. 2014;60(6):2027‐2039. - PMC - PubMed
    1. Kocabayoglu P, Lade A, Lee YA, et al. beta‐PDGF receptor expressed by hepatic stellate cells regulates fibrosis in murine liver injury, but not carcinogenesis. J Hepatol. 2015;63(1):141‐147. - PMC - PubMed
    1. Anthony B, Allen JT, Li YS, McManus DP. Hepatic stellate cells and parasite‐induced liver fibrosis. Parasit Vectors. 2010;3(1):60. - PMC - PubMed
    1. Duval F, Moreno‐Cuevas JE, González‐Garza MT, Rodríguez‐Montalvo C, Cruz‐Vega DE. Liver fibrosis and protection mechanisms action of medicinal plants targeting apoptosis of hepatocytes and hepatic stellate cells. Adv Pharmacol Sci. 2014;2014:373295. - PMC - PubMed
    1. Anthony B, Mathieson W, de Castro‐Borges W, Allen J. Schistosoma mansoni: egg‐induced downregulation of hepatic stellate cell activation and fibrogenesis. Exp Parasitol. 2010;124(4):409‐420. - PubMed

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