Knockdown of long noncoding RNA metastasis-associated lung adenocarcinoma transcript 1 protects against intracerebral hemorrhage through microRNA-146a-mediated inhibition of inflammation and oxidative stress

Figures & data

Figure 1. Long noncoding RNA MALAT1 was upregulated in intracerebral hemorrhage rats targeted miR-146a and miR-146a was downregulated. (a)The expression levels were detected using quantitative real-time PCR (qRT-PCR). (b) Binding sites between MALAT1 and miR-146a. (c) Dual-luciferase analysis proved that the miR-146a mimic repressed the luciferase activity of WT‑MALAT1, while no obvious effects on MUT‑MALAT1 activity. (d) The qRT-PCR assay was used to check miR-146a levels in intracerebral hemorrhage rats. Results are displayed as mean ± standard deviation, **p < 0.01, vs. sham; ##p < 0.01 vs. mimic control.

Figure 2. Inhibition of long noncoding RNA MALAT1 reduced neurological damage and brain edema in intracerebral hemorrhage rats. After 1 h of intracerebral hemorrhage (ICH) induction, control-shRNA, lncRNA MALAT1-shRNA, MALAT1-shRNA+inhibitor control, or MALAT1-shRNA+miR-146a inhibitor were transfected into ICH rats. (a) MALAT1 levels in different groups. (b) Expression of miR-146a in different groups. Neurobehavioral scores were assessed using the mNSS test in various groups after ICH for 1 h (c) and 3 d (d). (e) Brain water content was investigated using the wet/dry method in all groups. Data are displayed as means ± standard deviation, **p < 0.01 vs. sham; #, ##p < 0.05, 0.001 vs. ICH+control-shRNA; &, &&p < 0.05, 0.01 vs. ICH+MALAT1-shRNA+inhibitor control.

Figure 3. Downregulation of long noncoding RNA MALAT1 inhibited the pro‑inflammatory cytokine levels in intracerebral hemorrhage rats. ELISA was employed to determine the TNF-α and IL-1β levels in the serum and cerebrospinal fluid in rats of sham, ICH, ICH+control-shRNA, ICH+MALAT1-shRNA, ICH+MALAT1-shRNA+inhibitor control, and ICH+MALAT1-shRNA+miR-146a inhibitor groups. Data are expressed as the mean ± SD. **p < 0.01 vs. Sham; ##p < 0.001 vs. ICH+control-shRNA; &, &&p < 0.05, 0.01 vs. ICH+MALAT1-shRNA+inhibitor control.

Figure 4. Downregulation of long noncoding RNA MALAT1 suppressed oxidative stress in intracerebral hemorrhage rats.

SOD activity and MDA levels were measured in rat brain tissues from different groups. Results are displayed as mean ± SD. **p < 0.01 vs. Sham; ##p < 0.001 vs. ICH+control-shRNA; &&p < 0.01 vs. ICH+MALAT1-shRNA+inhibitor control.

Figure 5. Inhibition of long noncoding RNA MALAT1 inhibited neuronal apoptosis in intracerebral hemorrhage rats.

(A and B) The neuronal apoptosis was detected using flow cytometric analysis in the brain tissue of rats from sham, ICH, ICH + control-shRNA, ICH + MALAT1-shRNA, ICH + MALAT1-shRNA+inhibitor control, and ICH + MALAT1-shRNA+miR-146a inhibitor groups. (C) Caspase3 activity in the brain tissue of rats from sham, ICH, ICH + control-shRNA, ICH + MALAT1-shRNA, ICH + MALAT1-shRNA+inhibitor control, and ICH + MALAT1-shRNA+miR-146a inhibitor groups was calculated. Results are expressed as mean ± SD. **p < 0.01 vs. sham; ##p < 0.001 vs. ICH + control-shRNA; &&p < 0.01 vs. ICH + MALAT1-shRNA+inhibitor control.

Figure 6. Downregulation of long noncoding RNA MALAT1 suppressed the NF‑κB pathway in intracerebral hemorrhage rats.

Western blot assay was performed to determine the protein expression of p-p65 (A) and the ratio of p-p65/p65 (B) in the brain tissue of rats from the sham, ICH, ICH + control-shRNA, ICH + MALAT1-shRNA, ICH + MALAT1-shRNA+inhibitor control, and ICH + MALAT1-shRNA+miR-146a inhibitor groups. (C) The mRNA levels of p65 were determined using qRT-PCR. Results are shown as mean ± SD. **p < 0.01 vs. sham; ##p < 0.001 vs. ICH + control-shRNA; &&p < 0.01 vs. ICH + MALAT1-shRNA+inhibitor control.

Data Availability Statement

The datasets used and/or analyzed during the current study are available from the corresponding author upon reasonable request.