| Objective To analyze the molecular regulatory mechanism of tumor-associated long non-coding RNA expressed on chro-mosome 2 (TALNEC2) of long non-coding RNA (lncRNA) in the hypoxic-glucose deprivation model (OGD) of acute cerebral infarction in vitro.Methods The study was conducted from January 2021 to June 2022. Mouse microglia cell line BV2 was selected to establishthe OGD cell model, they were divided into Blank group (without any treatment), OGD group, OGD+si-NC group (transfected with si-NC for OGD treatment), and OGD+si-TALNEC2 group (transfected with si-TALNEC2 for OGD treatment). Real-time quantitative fluo-rescent PCR (qRT-PCR) was used to detect the expression of TALNEC2. Cell count kit (CCK-8), flow cytometry and enzyme-linked im-munosorbent assay (ELISA) were used to detect the changes of cell viability, apoptosis rate and inflammatory cytokines interleukin (IL)-1β, IL-18 and tumor necrosis factor α (TNF-α), respectively; Western blotting was used to detect proteins expression of NLR family Pyrin-domain protein 3 (NLRP3), apoptosis-associated speck-like protein (ASC), cleaved caspase-1, nuclear factor E2-related factor 2 (Nrf2), heme oxidase 1 (HO-1), and Zeste homolog enhancer 2 (EZH2); The binding of TALNEC2 to EZH2 was confirmed by RNA pull-down assay and RNA binding protein immunoprecipitation (RIP) assay.Results Compared with Blank group [1.02±0.03, 0.81±0.05, 1.29±0.06, 1.52±0.08, (4.72±1.09) %], the expression of TALNEC2 in OGD group was increased (4.18±0.15), the cell activity was decreasedat 24 h (0.46±0.05), 48 h (0.61±0.05), 72 h (0.75±0.02), and the apoptosis rate was increased (25.63±4.28) % (P<0.05); Compared with OGD group, the expression of TALNEC2 in OGD+si-TALNEC2 group decreased (1.16±0.09), the cell activity was increased at 24 h(0.65±0.07), 48 h (0.98±0.05) and 72 h (1.13±0.07), and the apoptosis rate was decreased (7.25±1.93) % (P<0.05). Compared with theBlank group [1.05±0.07, 1.04±0.05, 1.01±0.05, 1.02±0.04, 1.04±0.07, (47.21±5.01) ng/L, (38.26±4.13) ng/L, (48.22±7.15) ng/L, 1.06±0.03, 1.03±0.06], in the OGD group, the protein expressions of NLRP3 (2.69±0.15), ASC (3.11±0.18), cleaved caspase-1 (2.51±0.12), cytoplasmal Nrf2 (2.91±0.13), EZH2 (3.12±0.18) and inflammatory cytokines level of TNF-α (157.22±13.18) ng/L, IL-1β (177.43± 15.15) ng/L, IL-18 (2) 10.63±19.08) ng/L increased, and the protein expressions of nucleus Nrf2 (0.37±0.03) and HO-1 (0.27±0.04) de-creased (P<0.05); Compared with the OGD group, In OGD+si-TALNEC2 group, the protein expressions of NLRP3 (1.33±0.11), ASC (1.59±0.13), cleaved caspase-1 (1.42±0.09), cytoplasmal Nrf2 (1.39±0.06), EZH2 (1.47±0.09) and inflammatory cytokines level of TNF-α (74.38±6.24) ng/L, IL-1β (60.41±5.94) ng/L, IL-18 (86.25±9.76) ng/L decreased, and the protein expressions of nucleus Nrf2 (0.97± 0.05) and HO-1 (0.95±0.07) increased (P<0.05).Conclusion Knocking down lncRNA TALNEC2 can promote Nrf2 entry into the nu-cleus to up-regulate the expression of HO-1 by reducing binding to EZH2, and ultimately inhibit the activation of NLRP3 inflamma-some, thus reducing the injury of acute cerebral infarction. |
