Delta opioid peptide [D-ala2, D-leu5]-Enkephalin’s ability to enhance mitophagy via TRPV4 to relieve ischemia/reperfusion injury in brain microvascular endothelial cells

• DOI: 10.1136/svn-2023-003080
• Publication Date: 2024-05-02T17:00:13Z
• Authors: Zhongfang Deng; Xiaoyu Chen; Ran Zhang; Lingchao Kong; Yang Fang; Jizheng Guo; Bing Shen; Lesha Zhang
• Keywords: DADLE; TRPV4

Background Local brain tissue can suffer from ischaemia/reperfusion (I/R) injury, which lead to vascular endothelial damage. The peptide δ opioid receptor (δOR) agonist [D-ala2, D-leu5]-Enkephalin (DADLE) reduce apoptosis caused by acute I/R injury in microvascular cells (BMECs). Objective This study aims explore the mechanism DADLE enhances level of mitophagy BMECs upregulating expression transient potential vanilloid subtype 4 (TRPV4). Methods were extracted and made undergo oxygen-glucose deprivation/reoxygenation (OGD/R) accompanied DADLE. RNA-seq analysis revealed that induced increased TRPV4 expression. CCK-8 method was used assess cellular viability; quantitative PCR (qPCR) determine mRNA Drp1 ; western blot autophagy-related proteins; calcium imaging detect influx. Autophagosomes cells’ mitochondria observed using transmission electron microscopy. ELISA measure ATP content, a JC-1 fluorescent probe mitochondrial membrane potential. Results When compared with OGD/R group, OGD/R+DADLE group showed significantly enhanced TRPV4, Beclin-1, LC3-II/I, PINK1 Parkin; decreased p62 expression; marked rise influx; further increases mitophagy, an increase synthesis elevation These protective effects be blocked inhibitor HC067047 or RNAi TRPV4. Conclusion promote through improving function relieving injury.