Research Paper Volume 13, Issue 6 pp 8127—8145

Long noncoding RNA NKILA transferred by astrocyte-derived extracellular vesicles protects against neuronal injury by upregulating NLRX1 through binding to mir-195 in traumatic brain injury

NKILA delivered by astrocyte-derived EVs promotes neuron recovery by increasing NLRX1 expression via miR-195 in vitro. The injured neurons were treated with NKILA-EVs and vector-EVs. (A) observation of EVs released by astrocyte under an electron microscope (scale bar: 100 nm). (B) microscopic views of uptake of EVs by neurons (× 400). (C) expression of NKILA, miR-195 and NLRX1 in injured neurons measured by RT-qPCR. (D) protein expression of NLRX1 in injured neurons measured by Western blot analysis. (E) cell proliferation of injured neurons after treatment measured by EdU assay. (F) quantitative analysis for cell apoptosis of injured neurons after treatment measured by flow cytometry. (G) LDH content in injured neurons after treatment. (H) expression of Bcl-2, Bax and Caspase-3 in injured neurons after treatment measured by Western blot analysis. * p p t test was used for comparison between two groups. One-way ANOVA, followed by Tukey’s post-hoc test, was used for multi-group comparisons. The cell experiment was repeated three times.

Figure 7. NKILA delivered by astrocyte-derived EVs promotes neuron recovery by increasing NLRX1 expression via miR-195 in vitro. The injured neurons were treated with NKILA-EVs and vector-EVs. (A) observation of EVs released by astrocyte under an electron microscope (scale bar: 100 nm). (B) microscopic views of uptake of EVs by neurons (× 400). (C) expression of NKILA, miR-195 and NLRX1 in injured neurons measured by RT-qPCR. (D) protein expression of NLRX1 in injured neurons measured by Western blot analysis. (E) cell proliferation of injured neurons after treatment measured by EdU assay. (F) quantitative analysis for cell apoptosis of injured neurons after treatment measured by flow cytometry. (G) LDH content in injured neurons after treatment. (H) expression of Bcl-2, Bax and Caspase-3 in injured neurons after treatment measured by Western blot analysis. * p < 0.05 compared with neurons co-cultured with EV-free Ast-CM. # p < 0.05 compared with neurons co-cultured with Vector-EVs. All data were measurement data and expressed as mean ± standard deviation. Unpaired t test was used for comparison between two groups. One-way ANOVA, followed by Tukey’s post-hoc test, was used for multi-group comparisons. The cell experiment was repeated three times.