Regulations of IL-9 by the TGF-β/Smads signaling pathway in human Echinococcus granulosus infection

Abstract

Abstract:

Objective To examine mRNA expression of TGF-β, Smad3, Smad7, PU.1 and Interleukin-9 (IL-9) in livers of patients with cystic echinococcosis, in order to explore the regulations of Th9 cells by the TGF-β/Smad signaling in Echinococcus granulosus infection. Methods The liver mononuclear cells were isolated from liver tissues obtained during surgery from 30 patients with cystic hydatid disease and 20 patients with hepatic hemangioma (control group) in the First Affiliated Hospital of Xinjiang Medical University using the Percoll method. Real-time fluorescent quantitative PCR was performed to detect the mRNA expression of TGF-β, Smad3, Smad7, PU.1 and IL-9 in the mononuclear cells. The relationship between Smad3 mRNA expression and PU.1 mRNA expression was assessed by Spearman correlation analysis. The Smad3 inhibitor SIS3 or PBS was added to the mononuclear cells of the infected group, then PU.1 and IL-9 mRNA expression were determined by real-time fluorescent quantitative PCR, and IL-9 level in the supernatant of cell culture was assessed by ELISA. Results The relative mRNA expression of TGF-β, Smad3, PU.1 and IL-9 in CE patients were 2.55 ± 1.32, 2.26 ± 1.35, 1.71 ± 0.66 and 1.67 ± 0.75, respectively, all significantly higher than those in the control group (1.54 ± 0.76, 1.34 ± 0.51, 1.27 ± 0.62 and 1.01 ± 0.50, P < 0.05 or 0.01). The relative mRNA expression of Smad7 in CE patients (0.49 ± 0.37) was significantly lower than that in the control group (0.88 ± 0.50; P < 0.01). The Smad3 mRNA expression and PU.1 mRNA expression were positively correlated (r = 0.397, P < 0.05). After blocking the TGF-β/Smad signaling pathway by SIS3, the expression of PU.1 and IL-9 mRNA in liver mononuclear cells (1.00 ± 0.57, 0.85 ± 0.50) both decreased compared with the PBS treatment (1.46 ± 0.64, 1.32 ± 0.72) (P < 0.01). ELISA results showed that the IL-9 level in the supernatant of cell culture [(128.45 ± 41.97) pg/ml] also reduced compared with the PBS treatment [(166.64 ± 57.90) pg/ml; P < 0.05]. Conclusion The expression of TGF-β, Smad2, Smad3, PU.1 and IL-9 up-regulated in patients with cystic hydatid disease and the TGF-β/Smad pathway may regulate the expression of IL-9 after Echinococcus granulosus infection.

Key words: Echinococcus granulosus, Infection, TGF-β/Smad signal pathway;, PU.1, IL-9

CLC Number:

R532.32

Nan-nan PANG, Meng-ting AN, Feng-bo ZHANG, Xiao ZHAO, Fang YAN, Jian-bing DING. Regulations of IL-9 by the TGF-β/Smads signaling pathway in human Echinococcus granulosus infection[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2018, 36(5): 460-464.

Figures/Tables 2

References 21

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基因 Gene	引物序列（5′→3′） Primer sequences (5′→3′)	退火温度/℃ Tm/℃	长度/bp Length/bp
TGF-β1	F:ACACCAACTATTGCTTCAG	55.7	159
	R:TGTCCAGGCTCCAAATG
Smad3	F: GTGCTCCATCTCCTACTAC	56.5	183
	R: CCTCTTCCGATGTGTCTC
Smad7	F:TTTGTGTATTTATTTCTTTCTCTC	54.5	194
	R:CACTCTCGTCTTCTCCTC
Smad2	F: TTTGCTGCTCTTCTGGCTCA	60	179
	R: AACCACTGTAGAGGTCCATTCA
PU.1	F: GGAGCCCGGCTGGATGTTAC	60.04	79
	R: CACCAGGTCTTCTGATGGCTGA
IL-9	F: CTCTAGCAGTCCACTTCACCAA	59.3	112
	R: ACAGCATGGGTCTGTCTTCT
β-Actin	F: TAGGCGGACTGTTACTGAGC	60	233
	R: TGCTCCAACCAACTGCTGTC