Safflor Yellow B Attenuates Ischemic Brain Injury via Downregulation of Long Noncoding AK046177 and Inhibition of MicroRNA-134 Expression in Rats

Figure 2

Effect of safflor yellow B on TUNEL-positive cells, cell viability, and apoptosis. Rats were divided into six groups: sham, ischemia/reperfusion (I/R), AK046177 siRNA, AK046177 siRNA+miR-134 agomir, SYB, and SYB+miR-134 agomir. (a) Representative images showing TUNEL-positive cells of cerebral cortex in different groups (×200 magnification; 1, 2, 3, 4, 5, and 6 represent sham, ischemia/reperfusion (I/R), AK046177 siRNA, AK046177 siRNA+miR-134 agomir, SYB, and SYB+miR-134 agomir, respectively). (b) Apoptotic (TUNEL-positive) cells were detected, . Primary fetal cortical cells were seeded in 96-well and 6-well plates and divided into six groups: control, OGD/R, AK046177 siRNA, AK046177 siRNA+miR-134 agomir, SYB, and SYB+miR-134 agomir. Apart from the control group, all cells in the other groups were cultured in glucose-free DMEM and hypoxic conditions (1% O₂/94% N₂/5% CO₂) at 37°C for 4 h. Thereafter, all groups’ media were replaced with normal DMEM, and culturing continued for 20 h of reoxygenation under normoxic conditions (95% air/5% CO₂). The cells were pretreated with SYB, AK046177, and miR-134 agomir before being exposed to OGD/R. Cell viability was detected by the MTT method. Cell apoptosis was analyzed using flow cytometry. (c1)–(c6) represent control, OGD/R, AK046177 siRNA, AK046177 siRNA+miR-134 agomir, SYB, and SYB+miR-134 agomir, respectively. (d) represents cell apoptosis (). (e) represents cell viability (). Data are presented as . One-way ANOVA test was used to determine statistical significance. vs. the sham group or the control group, ^# or ^##vs. the I/R group or the OGD/R group, ^Δ or ^ΔΔvs. the AK046177 siRNA group, and ⁺ or ⁺⁺vs. the SYB group.

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