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MicroRNA-19b-3p Modulates Japanese Encephalitis Virus-Mediated Inflammation via Targeting RNF11.

Ashraf U, Zhu B, Ye J, Wan S, Nie Y, Chen Z, Cui M, Wang C, Duan X, Zhang H, Chen H, Cao S - J. Virol. (2016)

Bottom Line: Japanese encephalitis virus (JEV) can invade the central nervous system and consequently induce neuroinflammation, which is characterized by profound neuronal cell damage accompanied by astrogliosis and microgliosis.The pathological features of JEV-induced encephalitis are inflammatory reactions and neurological diseases resulting from glia activation.The present study reveals that miR-19b-3p targets ring finger protein 11 in glia and promotes inflammatory cytokine production by enhancing nuclear factor kappa B activity in these cells.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, Hubei, People's Republic of China Laboratory of Animal Virology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei, People's Republic of China The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan, Hubei, People's Republic of China Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, Hubei, People's Republic of China.

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Regulation of JEV-induced production of inflammatory cytokines by miR-19b-3p is achieved through RNF11. (A) U251 cells were transfected with siRNF11 or nonspecific control siRNA (siNC) (final concentration, 50 nM) for 24 h, and then RNF11 protein levels were measured by immunoblotting. Protein levels were quantified by immunoblot scanning and normalized to the amount of GAPDH expression. (B) U251 cells were cotransfected with miR-19b-3p inhibitors or control oligonucleotides and siRNF11 or a nonspecific control siRNA (final concentration, 50 nM) for 24 h and then infected with JEV at an MOI of 5 for 36 h. The levels of miR-19b-3p were analyzed by quantitative real-time PCR and normalized to the U6 level. **, P < 0.01; ***, P < 0.001. (C) The protein levels of TNF-α, IL-6, and IL-1β were analyzed by ELISA. Data represent means ± SD from three independent experiments performed in duplicate. *, P < 0.05; **, P < 0.01; ***, P < 0.001. CCL5 mRNA levels were determined with quantitative real-time PCR and normalized to the expression of β-actin in each sample. Data represent means ± SD from three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001.
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Figure 6: Regulation of JEV-induced production of inflammatory cytokines by miR-19b-3p is achieved through RNF11. (A) U251 cells were transfected with siRNF11 or nonspecific control siRNA (siNC) (final concentration, 50 nM) for 24 h, and then RNF11 protein levels were measured by immunoblotting. Protein levels were quantified by immunoblot scanning and normalized to the amount of GAPDH expression. (B) U251 cells were cotransfected with miR-19b-3p inhibitors or control oligonucleotides and siRNF11 or a nonspecific control siRNA (final concentration, 50 nM) for 24 h and then infected with JEV at an MOI of 5 for 36 h. The levels of miR-19b-3p were analyzed by quantitative real-time PCR and normalized to the U6 level. **, P < 0.01; ***, P < 0.001. (C) The protein levels of TNF-α, IL-6, and IL-1β were analyzed by ELISA. Data represent means ± SD from three independent experiments performed in duplicate. *, P < 0.05; **, P < 0.01; ***, P < 0.001. CCL5 mRNA levels were determined with quantitative real-time PCR and normalized to the expression of β-actin in each sample. Data represent means ± SD from three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Mentions: To determine whether the observed effects of miR-19b-3p on inflammatory cytokine production in response to JEV infection were, at least partially, mediated through RNF11, we analyzed the effects of silencing RNF11 expression by siRNA in U251 cells. We confirmed that the siRNA significantly inhibited RNF11 protein expression in U251 cells (Fig. 6A).


MicroRNA-19b-3p Modulates Japanese Encephalitis Virus-Mediated Inflammation via Targeting RNF11.

Ashraf U, Zhu B, Ye J, Wan S, Nie Y, Chen Z, Cui M, Wang C, Duan X, Zhang H, Chen H, Cao S - J. Virol. (2016)

Regulation of JEV-induced production of inflammatory cytokines by miR-19b-3p is achieved through RNF11. (A) U251 cells were transfected with siRNF11 or nonspecific control siRNA (siNC) (final concentration, 50 nM) for 24 h, and then RNF11 protein levels were measured by immunoblotting. Protein levels were quantified by immunoblot scanning and normalized to the amount of GAPDH expression. (B) U251 cells were cotransfected with miR-19b-3p inhibitors or control oligonucleotides and siRNF11 or a nonspecific control siRNA (final concentration, 50 nM) for 24 h and then infected with JEV at an MOI of 5 for 36 h. The levels of miR-19b-3p were analyzed by quantitative real-time PCR and normalized to the U6 level. **, P < 0.01; ***, P < 0.001. (C) The protein levels of TNF-α, IL-6, and IL-1β were analyzed by ELISA. Data represent means ± SD from three independent experiments performed in duplicate. *, P < 0.05; **, P < 0.01; ***, P < 0.001. CCL5 mRNA levels were determined with quantitative real-time PCR and normalized to the expression of β-actin in each sample. Data represent means ± SD from three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 6: Regulation of JEV-induced production of inflammatory cytokines by miR-19b-3p is achieved through RNF11. (A) U251 cells were transfected with siRNF11 or nonspecific control siRNA (siNC) (final concentration, 50 nM) for 24 h, and then RNF11 protein levels were measured by immunoblotting. Protein levels were quantified by immunoblot scanning and normalized to the amount of GAPDH expression. (B) U251 cells were cotransfected with miR-19b-3p inhibitors or control oligonucleotides and siRNF11 or a nonspecific control siRNA (final concentration, 50 nM) for 24 h and then infected with JEV at an MOI of 5 for 36 h. The levels of miR-19b-3p were analyzed by quantitative real-time PCR and normalized to the U6 level. **, P < 0.01; ***, P < 0.001. (C) The protein levels of TNF-α, IL-6, and IL-1β were analyzed by ELISA. Data represent means ± SD from three independent experiments performed in duplicate. *, P < 0.05; **, P < 0.01; ***, P < 0.001. CCL5 mRNA levels were determined with quantitative real-time PCR and normalized to the expression of β-actin in each sample. Data represent means ± SD from three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001.
Mentions: To determine whether the observed effects of miR-19b-3p on inflammatory cytokine production in response to JEV infection were, at least partially, mediated through RNF11, we analyzed the effects of silencing RNF11 expression by siRNA in U251 cells. We confirmed that the siRNA significantly inhibited RNF11 protein expression in U251 cells (Fig. 6A).

Bottom Line: Japanese encephalitis virus (JEV) can invade the central nervous system and consequently induce neuroinflammation, which is characterized by profound neuronal cell damage accompanied by astrogliosis and microgliosis.The pathological features of JEV-induced encephalitis are inflammatory reactions and neurological diseases resulting from glia activation.The present study reveals that miR-19b-3p targets ring finger protein 11 in glia and promotes inflammatory cytokine production by enhancing nuclear factor kappa B activity in these cells.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, Hubei, People's Republic of China Laboratory of Animal Virology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei, People's Republic of China The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan, Hubei, People's Republic of China Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, Hubei, People's Republic of China.

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Related in: MedlinePlus