Figures & data
Figure 1. Overexpression of miR-193b-3p alleviated pathological damage in I/R rats. The miR-193b-3p agomir or miR-193b-3p antagomir was injected into the lateral ventricle of rats to induce overexpression or downregulation of miR-193b-3p. The rat brain I/R injury model was established using the suture method. After 2 h of ischemia and 24 h of reperfusion, the rat brain tissues were collected for relevant detection. (A) HE staining was used to observe the changes of neuron structure and TTC staining was used to analyze the infarct volume of rat brain; (B) TUNEL staining was used to detect the level of neuronal apoptosis in rats; (C) The level of miR-193b-3p in rat brain tissue was detected by RT-qPCR; Measurement data were expressed as mean ± standard deviation, N = 6, one-way ANOVA was used for data comparison among groups, and Tukey’s multiple test was used for post hoc test. P was obtained from a bilateral test, ** P < 0.01, *** P < 0.001.
Figure 2. Overexpression of miR-193b-3p inhibits oxidative stress and mitochondrial apoptosis of neurons in the cerebral cortex of rats. The miR-193b-3p agomir was injected into the lateral ventricle of rats to induce the overexpression of miR-193b-3p, and the rat brain I/R injury model was established by suture method. After 2 h of ischemia and 24 h of reperfusion, the rat brain tissues were collected for relevant detection. (A) DCFH-DA fluorescent probe was used to detect ROS levels in neurons of rat brain tissue; (B) SOD level of neurons in rat brain tissue was detected by the kit; (C) MDA level of neurons in rat brain tissue was detected by the kit; (D) JC-1 fluorescence probe was used to detect MMP of neuron mitochondria in rat brain; (E) The level of mitochondrial ATP in rat brain tissue was detected by kits. Measurement data were expressed as mean ± standard deviation, N = 6, one-way ANOVA was used for data comparison among groups, and Tukey’s multiple test was used for the post hoc test. P was obtained from a bilateral test, ** P < 0.01, *** P < 0.001.
Figure 3. miR-193b-3p targeted SIAH1. (A) The targeted binding sites of miR-193b-3p and SIAH1 were predicted by TargetScan; (B) The targeting relationship between mir193B-3p and SIAH1 was verified by dual-luciferase reporter assay; (C) The mRNA and protein levels of SIAH1 in brain tissue of rats in different groups were detected by RT-qPCR and WB; Measurement data were expressed as mean ± standard deviation, N = 6, one-way ANOVA was used for data comparison among groups, and Tukey’s multiple test was used for the post hoc test. P was obtained from a bilateral test, ** P < 0.01, *** P< 0.001.
Figure 4. Overexpression of SIAH1 antagonizes the protective effect of overexpression of miR-193b-3p on cerebral I/R injury in rats. The SIAH1 was overexpressed in rats by injecting the SIAH1 lentivirus. (A) The level of SIAH1 was detected by RT-qPCR and WB. (B) DCFH-DA fluorescent probe was used to detect ROS levels in rat brain tissue; (C) SOD level of neurons in rat brain tissue was detected by the kit; (D) MDA level of neurons in rat brain tissue was detected by the kit; (E) JC-1 fluorescence probe was used to detect MMP of neuron mitochondria in rat brain tissue. (F) The kit was used to detect the level of mitochondrial ATP of neurons in rat brain tissue; (G) HE staining and TTC staining were used to detect the neuronal damage and cerebral infarct volume of rat brain tissue; (H) TUNEL staining was used to detect neuronal apoptosis in rats. Measurement data were expressed as mean ± standard deviation, N = 6, one-way ANOVA was used for data comparison among groups, and Tukey’s multiple test was used for the post hoc test. P was obtained from a bilateral test, ** P < 0.01, *** P < 0.001.
Figure 5. miR-193b-3p inhibits the activation of the JNK pathway by targeting SIAH1. (A-B): The phosphorylation level of JNK1/2/3 in the brain tissues of rats in different groups was detected by WB. Measurement data were expressed as mean /2/3 in the brain tissue. N = 6, one-way ANOVA was used for data comparison among groups, and Tukey’s multiple test was used for the post hoc test. P was obtained from a bilateral test, ** P < 0.01, *** P < 0.001.
