Limits...
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.

Show MeSH

Related in: MedlinePlus

miR-19b-3p activates the NF-κB pathway in JEV-infected astrocytes. (A and B) U251 cells were transfected with miR-19b-3p mimics, inhibitors, or their control oligonucleotides (final concentration, 50 nM) for 24 h and then either left uninfected or infected with JEV at an MOI of 5 for 36 h. The cytosolic extracts (upper panel) and nuclear extracts (lower panel) were isolated and subjected to immunoblotting with antibodies against RNF11, NF-κB p65, lamin A, and GAPDH. Lamin A was used as a marker for nuclei. GAPDH and lamin A were used as the loading controls. Protein levels were quantified by immunoblot scanning and normalized to the amount of GAPDH or lamin A expression. *, P < 0.05; **, P < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4836334&req=5

Figure 7: miR-19b-3p activates the NF-κB pathway in JEV-infected astrocytes. (A and B) U251 cells were transfected with miR-19b-3p mimics, inhibitors, or their control oligonucleotides (final concentration, 50 nM) for 24 h and then either left uninfected or infected with JEV at an MOI of 5 for 36 h. The cytosolic extracts (upper panel) and nuclear extracts (lower panel) were isolated and subjected to immunoblotting with antibodies against RNF11, NF-κB p65, lamin A, and GAPDH. Lamin A was used as a marker for nuclei. GAPDH and lamin A were used as the loading controls. Protein levels were quantified by immunoblot scanning and normalized to the amount of GAPDH or lamin A expression. *, P < 0.05; **, P < 0.01.

Mentions: It has been well established that translocation of NF-κB from the cytoplasm to the nucleus is a key determinant of NF-κB activation (36, 48). Therefore, it was of interest to evaluate the effect of miR-19b-3p on NF-κB activation in JEV-infected astrocytes. Nuclear translocation of NF-κB (p65) was detected with immunoblotting. Transfection of miR-19b-3p mimics increased the translocation of NF-κB from the cytoplasm to the nucleus (Fig. 7A). In contrast, treatment of cells with miR-19b-3p inhibitors significantly inhibited the nuclear translocation of NF-κB in JEV-infected U251 cells (Fig. 7B). Thus, these findings demonstrated that miR-19b-3p appears to regulate inflammatory cytokine production by enhancing the activation of NF-κB signaling in JEV-infected astrocytes.


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)

miR-19b-3p activates the NF-κB pathway in JEV-infected astrocytes. (A and B) U251 cells were transfected with miR-19b-3p mimics, inhibitors, or their control oligonucleotides (final concentration, 50 nM) for 24 h and then either left uninfected or infected with JEV at an MOI of 5 for 36 h. The cytosolic extracts (upper panel) and nuclear extracts (lower panel) were isolated and subjected to immunoblotting with antibodies against RNF11, NF-κB p65, lamin A, and GAPDH. Lamin A was used as a marker for nuclei. GAPDH and lamin A were used as the loading controls. Protein levels were quantified by immunoblot scanning and normalized to the amount of GAPDH or lamin A expression. *, P < 0.05; **, P < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4836334&req=5

Figure 7: miR-19b-3p activates the NF-κB pathway in JEV-infected astrocytes. (A and B) U251 cells were transfected with miR-19b-3p mimics, inhibitors, or their control oligonucleotides (final concentration, 50 nM) for 24 h and then either left uninfected or infected with JEV at an MOI of 5 for 36 h. The cytosolic extracts (upper panel) and nuclear extracts (lower panel) were isolated and subjected to immunoblotting with antibodies against RNF11, NF-κB p65, lamin A, and GAPDH. Lamin A was used as a marker for nuclei. GAPDH and lamin A were used as the loading controls. Protein levels were quantified by immunoblot scanning and normalized to the amount of GAPDH or lamin A expression. *, P < 0.05; **, P < 0.01.
Mentions: It has been well established that translocation of NF-κB from the cytoplasm to the nucleus is a key determinant of NF-κB activation (36, 48). Therefore, it was of interest to evaluate the effect of miR-19b-3p on NF-κB activation in JEV-infected astrocytes. Nuclear translocation of NF-κB (p65) was detected with immunoblotting. Transfection of miR-19b-3p mimics increased the translocation of NF-κB from the cytoplasm to the nucleus (Fig. 7A). In contrast, treatment of cells with miR-19b-3p inhibitors significantly inhibited the nuclear translocation of NF-κB in JEV-infected U251 cells (Fig. 7B). Thus, these findings demonstrated that miR-19b-3p appears to regulate inflammatory cytokine production by enhancing the activation of NF-κB signaling in JEV-infected astrocytes.

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.

Show MeSH
Related in: MedlinePlus