Para‑hydroxybenzaldehyde against transient focal cerebral ischemia in rats via mitochondrial preservation

Abstract

Gastrodia elata (GE) Blume has been widely used for thousands of years to treat various central and peripheral nervous disorders. P-hydroxybenzaldehyde (PHBA) is a chemical component of GE. However, its role and mechanism in transient focal cerebral ischemia remain unclear. The present study aimed to investigate the protective effect of PHBA on middle cerebral artery occlusion (MCAO) rats. A total of 56 Sprague-Dawley male rats were randomly divided into control, model, PHBA-high dose (PHBA-H) and PHBA-low dose (PHBA-L) groups. The MCAO injury model was replicated in all rats except for the control group. In the control group, only the right common carotid artery was isolated without embolization. After treatment with PHBA, the protective effects (neurological deficit score, cerebral index, weight and cerebral infarct) were analyzed. Western blotting was performed to estimate the protein levels of Bcl-2, Bax and Caspase-3. Apoptotic cells were detected using hematoxylin-eosin staining and TUNEL immunofluorescence assay. Mitochondrial oxidative stress indicators, including reactive oxygen species (ROS), malondialdehyde (MDA) and total superoxide dismutase (T-SOD), while dysfunction indicators, including mitochondrial permeability transition pore (MPTP), ATP and cytochrome C oxidase, were measured using commercial kits. The ultrastructure of mitochondria was observed under an electron microscope. Once the model was successful established, the rats in the MCAO group suffered neurological damage (P<0.001), increased cerebral index (P<0.001), decreased body weight (P<0.001) and had severe cerebral infarction (P<0.001). Moreover, the number of apoptotic cells and the levels of ROS (P<0.001) and MDA (P<0.05) in mitochondria and the protein levels of Bax (P<0.001) and cleaved caspase-3 (P<0.001) were increased. The activities of T-SOD (P<0.001) and cytochrome C oxidase (P<0.001) in the mitochondria, ATP content (P<0.05) and Bcl-2 protein level (P<0.001) decreased, MPTP was stimulated to open and mitochondrial structures were damaged (P<0.001). PHBA treatment resulted in a decrease of the neurological deficit score (PHBA-H 24 h, P<0.001; PHBA-H 6 h and PHBA-L 24 h, P<0.01; PHBA-L 6 h, P<0.05), apoptotic cell number (P<0.001), mitochondrial ROS (P<0.001) and MPTP opening (P<0.001), Bax (P<0.01, P<0.001) and cleaved caspase-3 protein expression (P<0.001) in rats. And the expression of Bcl-2 protein (P<0.001) was increased. In addition, the cerebral index (P<0.05), weight loss (P<0.05), infarction rate (P<0.01) and MDA content (P<0.001) were decrease in the PHBA-H group. The level of ATP (P<0.05) and cytochrome C oxidase (P<0.05) and T-SOD activity (P<0.05) of PHBA-H group rats increased, but no significant difference was observed in the PHBA-L group. Overall, PHBA had a protective effect on transient focal cerebral ischemia in normal rats, regulated the expression of Bcl-2, Bax and cleaved caspase-3 proteins and improved the oxidative stress and dysfunction of mitochondria.

P‑hydroxybenzyl alcohol ameliorates neuronal cerebral ischemia‑reperfusion injury by activating mitochondrial autophagy through SIRT1

“…1B ). Our previous study examined a pHBA derivative, p-hydroxybenzaldehyde, which comprised a benzene ring and an aldehyde group with a hydroxyl group ( 20 ). Following drug administration, MCAO modelling and reperfusion, the neurological deficit score of rats was evaluated.…”

“…Our previous study showed that p-hydroxybenzaldehyde, a component of G. elata , can protect MCAO/R rats from oxidation and apoptosis ( 20 ). Compared with the MCAO/R group, p-hydroxybenzaldehyde treatment reduced mitochondrial ROS and malondialdehyde contents, reduced mPTP opening, and decreased Bax and caspase-3 protein expression levels.…”

“…Compared with the MCAO/R group, p-hydroxybenzaldehyde treatment reduced mitochondrial ROS and malondialdehyde contents, reduced mPTP opening, and decreased Bax and caspase-3 protein expression levels. In addition, ATP content, cytochrome c oxidase and total superoxide dismutase activities, as well as protein expression of Bcl2 all increased ( 20 ). Similarly, results from the present study demonstrated that pHBA treatment increased ATP level, mPTP opening and Bcl2 expression, whereas mPTP opening and Bax and cleaved caspase-3 expression levels were decreased compared with the untreated MCAO/R model group.…”

“…Gastrodia elata is an herbal medicine commonly used in Asian countries to treat depression, epilepsy and other neurological diseases ( 19 ). Our previous study demonstrated that a component of G. elata , p-hydroxybenzaldehyde, can inhibit apoptosis and improve brain injury in MCAO/R rats ( 20 ). In addition, another component, p-hydroxybenzyl alcohol (pHBA), was found to have pharmacological effects such as anti-cerebral ischemic injury, anti-platelet aggregation and anti-inflammation ( 21 ).…”

Based on network pharmacology and molecular docking to predict the mechanism of TMDZ capsule in the treatment of IS

“…At present, it has been reported that sitosterol, vanillin, carotene, gastrodin, p-hydroxybenzaldehyde and other components can reduce blood cholesterol, inhibit the formation of cerebral arteriosclerosis, inhibit the excessive accumulation of cholesterol in nerve cells, anti-apoptosis of nerve cells, maintain the homeostasis of neurogliocyte and protect the integrity of blood-brain barrier. [28][29][30][31][32] In the treatment of IS, these components can inhibit the accumulation of cholesterol in nerve cells, regulate the cholesterol transport function of cholesterol transporter NPC1L1 on the cell membrane, reduce the expression levels of Cathepsin B and Caspase-3 and inhibit the apoptosis pathway of Cathepsin B-caspase. kaempferol, curcumin, ferulic acid, epicatechin and other components regulate the phosphorylation level of Akt and Src proteins, [33] prolong the reaction time of PT and APTT, [34] increase the activity of plasma t-PA, [35] decrease the activity of PAI-1, [36] increase the content of intracellular CGMP, [37] antagonistic endothelin, [38] blocking calcium channels, [39] dilating blood vessels, [40] reducing blood viscosity, reducing platelet aggregation, [41] restoring damaged endothelial tissue, [42] enhancing the activity of plasma anticoagulant system, [43] enhancing the activity of fibrinolytic system, [44] improving blood perfusion, [45] thrombolysis, inhibiting thrombosis, activating platelets, [46] reducing the release of platelet inflammatory factors, [47] reducing the density and coverage of platelet accumulation in cerebral thrombosis and other ways to anti-oxidation, anti-inflammation, anti-tumor and neuroprotection and other pharmacological effects.…”