Adenosine deaminase acting on RNA 2 provides neuroprotection by activating Kv1.1 channels in a rat epilepsy model

Figures

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  Figure 1:. Hippocampal ADAR2 and Kv1.1 expression changes. (a and b) Immunofluorescence results for ADAR2 in the sham, model, and oe-ADAR2 groups (n = 6). (c-e) Western blot and quantitative analyses of ADAR2 and Kv1.1 protein expressions (n = 6). (f and g) ADAR2 and Kv1.1 mRNA expressions detected via qRT-PCR (n = 3), magnification: ×200, scale: 100 mm. (**P < 0.01; ***P < 0.001, ****P < 0.0001. Sham: Sham, oe: Overexpression, ADAR2: Adenosine deaminase acting on RNA 2, GAPDH: Glyceraldehyde-3-phosphate dehydrogenase, Kv1.1: potassium voltage-gated channel subfamily A member 1, qRT-PCR: Quantitative reverse transcription polymerase chain reaction, DAPI: 4¢,6-Diamidino-2-phenylindole).

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  Figure 2:. Effects of ADAR2 overexpression on hippocampal neurons. (a) Hematoxylin and eosin (HE) staining, (b) Nissl staining; (c and d) immunohistochemical staining and quantitative analysis of neurons in the sham, model, and oe-ADAR2 groups (n = 6). Magnification: ×200; scale: 100 mm. ***P < 0.001. (Sham: Sham, oe: Overexpression, ADAR2: Adenosine deaminase acting on RNA 2, GAPDH: Glyceraldehyde-3-phosphate dehydrogenase, Kv1.1: Potassium voltage-gated channel subfamily A member 1, HE: Hematoxylin and eosin).

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  Figure 3:. Effect of ADAR2 overexpression on hippocampal Bax, Bcl-2, and cleaved caspases-3/7 expression. (a) Western blot bands of Bax, Bcl-2, and cleaved caspases-3/7. (b-e) Western blot analysis of Bax, Bcl-2, and cleaved caspases-3/7 (n = 3). **P < 0.01; ***P < 0.001, ****P < 0.0001. (Sham: Sham, oe: Overexpression, ADAR2: Adenosine deaminase acting on RNA 2, GAPDH: Glyceraldehyde-3-phosphate dehydrogenase, Kv1.1: Potassium voltage-gated channel subfamily A member 1, Bcl-2: B-cell leukemia/lymphoma 2 protein, Bax: Bcl-2-associated X protein).

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  Figure 4:. Effects of ADAR2 overexpression on the expressions of Kv1.1, Bax, Bcl-2, and cleaved caspases-3/7 in SH-SY5Y cells from the Con, EP, and oe-ADAR2 groups. (a-c) Western blot bands and quantitative analysis of ADAR2 and Kv1.1. (d-h) Western blot bands and quantitative analysis of Bax, Bcl-2, and cleaved caspases-3/7 (n = 3). (*P < 0.05, **P < 0.01; ***P < 0.001, ****P < 0.0001. Con: Control, oe: Overexpression, EP: Epilepsy, ADAR2: Adenosine deaminase acting on RNA 2, GAPDH: Glyceraldehyde-3-phosphate dehydrogenase, Kv1.1: Potassium voltage-gated channel subfamily A member 1, Bcl-2: B-cell leukemia/lymphoma 2 protein, Bax: Bcl-2-associated X protein.).

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  Figure 5:. Effects of ADAR2 overexpression on the expressions of Kv1.1, Bax, Bcl-2, and cleaved caspases 3 and 7 in SH-SY5Y cells from the Con, EP, oe-ADAR2, and oe-ADAR2+si-Kv1.1 groups. (a) Results on the knockdown efficiency of Kv1.1 by qRT-PCR. (b-d) Western blot bands and quantitative analysis of Kv1.1 and ADAR2. (e-i) Western blot bands and quantitative analysis of Bax, Bcl-2, and cleaved caspases-3/7 (n=3). (*P < 0.05, **P < 0.01; ***P < 0.001, ****P < 0.0001, ns: No significance. Si-NC: Si-negative control group, Con: Control, oe: Overexpression, EP: Epilepsy, ADAR2: Adenosine deaminase acting on RNA 2, GAPDH: Glyceraldehyde-3-phosphate dehydrogenase, Kv1.1: Potassium voltage-gated channel subfamily A member 1, qRT-PCR: Quantitative reverse transcription polymerase chain reaction, Bcl-2: B-cell leukemia/lymphoma 2 protein, Bax: Bcl-2-associated X protein).