Devil or angel: two roles of carbon monoxide in stroke.

Stroke is one of the most important acute diseases that endanger human health and result in death, including acute cerebral hemorrhage and acute cerebral ischemia. Acute onset is its most prominent feature. Carbon monoxide (CO) is a colorless and odorless gas existing at room temperature. It is not only a common air pollutant, but also has been found to be closely related to stroke. A large amount of exogenous CO has an important impact on the incidence and prognosis of stroke, while endogenous CO as a gas signal also has an important impact on neuroprotection after stroke. Both low-dose CO inhalation and CO-releasing molecule-3 (a molecule that emits CO) treatment have shown the benefits of stroke, and perhaps the role of CO in stroke is one of the key areas for future research.

Spectroscopic methodologies and molecular docking studies on the interaction of antimalarial drug piperaquine and its metabolites with human serum albumin.

Artemisinin-based combination therapy is widely used for the treatment of uncomplicated Plasmodium falciparum malaria, and piperaquine (PQ) is one of the important partner drugs. During the biotransformation of PQ, M1 (N-oxidation product), M2 (N-oxidation product), M3 (carboxylic acid product), M4 (N-dealkylation product), and M5 (N-oxidated product of M4) are formed by cytochrome P450 pathways. Despite decades of clinical use, the interactions between PQ and its main metabolites (PQs) with human serum albumin (HSA) have not been reported. In the present study, the binding of PQs with HSA under physiological conditions was investigated systematically through fluorescence, circular dichroism (CD) spectroscopy, and molecular docking methods. The experimental results show that the intrinsic fluorescence quenching of HSA was induced by those compounds resulting from the formation of stable HSA-compound complexes. The main forces involved in the interactions between PQ, M1, and M2 which bind to HSA were hydrogen s and van der Waals forces, while the interactions of M3, M4, and M5 were driven by hydrophobic forces. The main binding sites of the compounds to HSA were also examined by classical fluorescent marker experiments and molecular docking studies. Binding constants (Kb) revealed that the affinities of the PQ, M1, M2, M3, and M4 to HSA were stronger than that of M5. Additionally, the binding rates of PQs with HSA were determined by ultrafiltration methods. Consistent with the binding constant results, the binding rate of M5 was lower than the binding rates of PQ, M1, M2, M3, and M4. Furthermore, PQs binding to HSA led to conformational and structural alterations of HSA, as revealed by multi-spectroscopic studies. In order to investigate one possible mechanism by which PQs inhibit the growth of malaria-causing Plasmodium parasites, 1H NMR spectroscopy was performed to investigate the interaction of the PQs with heme. This study is beneficial to enhance our understanding of the ecotoxicology and environmental behaviors of PQ and its metabolites.

Inhibition of Lats1/p-YAP1 pathway mitigates neuronal apoptosis and neurological deficits in a rat model of traumatic brain injury.

AIMS: To investigate the roles of Lats1/p-YAP1 pathway in TBI-induced neuronal apoptosis and neurological deficits in rats. RESULTS: We found that Lats1 and YAP1 were expressed in cerebral cortex neurons of Sprague-Dawley rats, and the phosphorylation levels of Lats1 and YAP1 in injured regions were significantly increased after TBI. Furthermore, inhibition of Lats1 not only decreased the level of p-YAP1, but also attenuated neuronal apoptosis and neurological impairment. CONCLUSIONS: Our work demonstrates that inhibition of Lats1/p-YAP1 pathway mitigates neuronal apoptosis and neurological deficits in a rat model of TBI.

Investigation of the active components in Tripterygium wilfordii leading to its acute hepatotoxicty and nephrotoxicity.

ETHNOPHARMACOLOGICAL RELEVANCE: The traditional herbal medicine Tripterygium wilfordii Hook. f. (TW) has been widely used for the treatment of rheumatoid arthritis and autoimmune disease in the clinic. However, adverse reactions of TW including hepatotoxicity and nephrotoxicity have been frequently reported. Terpenes and alkaloids are among the most important active components in TW. Triptolide (TP), a major terpene in TW, has been found to induce toxicity, and metabolic pathways could lead to detoxification of TP. In this study, whether other major terpenes or alkaloids in TW contribute to its toxicity was investigated. The role of metabolic eliminations in their potential detoxification process was also evaluated. MATERIALS AND METHODS: The toxicity of TW and its five major active components (one terpene and four alkaloids) in mice was evaluated in terms of mortality and blood biochemical levels (ALT, AST, BUN and CREA). TP was used as a positive control. Metabolic pathways leading to potential detoxification of TW or its two representative components (triptonide and wilforgine) were evaluated in glutathione (GSH)-depleted (treated with L-buthionine-S,R-sulfoxinine, BSO) and aminobenzotriazole (ABT; a nonspecific inhibitor for P450s)-treated mice. RESULTS: In normal mice, the major metabolic pathways for the terpene compounds TP and triptonide (TN) were hydroxylation and cysteine conjugation, and the alkaloid wilforgine (WG) mainly underwent oxidative metabolism and hydrolysis. In ABT/BSO-treated mice, the hydroxylated metabolites of TP, TN and WG were found at a lower level than normal mice, and the level of cysteine conjugates of TN increased probably due to the stress response. Compared with normal mice, mortality and levels of ALT (but not BUN) were significantly higher (P<0.01) in TW (or TP)-treated mice (1.2 mg kg(-1)), indicating the acute toxicity (may not nephrotoxicity) of TW and its active component TP. Pretreatment with ABT and/or BSO increased the acute toxicity (including hepatotoxicity and nephrotoxicity) caused by TW or TP. No significant toxicity was found for TN or four alkaloids in normal mice or ABT/BSO-treated mice. CONCLUSIONS: TP was probably the main contributor to the toxicity of TW, and the terpene TN and alkaloids in TW may be of no toxicological concern at dosage levels up to 20-fold of the therapeutic dose. Metabolic eliminations to less reactive metabolites implied a high potential for detoxification of TW, and caution should be taken for TW clinical use during co-administration with other CYP inhibitors or GSH-depleting agents.

Potential contribution of hypoxia-inducible factor-1α, aquaporin-4, and matrix metalloproteinase-9 to blood-brain barrier disruption and brain edema after experimental subarachnoid hemorrhage.

The current research aimed to investigate the role of hypoxia-inducible factor-1α (HIF-1α), aquaporin-4 (AQP-4), and matrix metalloproteinase-9 (MMP-9) in blood-brain barrier (BBB) dysfunction and cerebral edema formation in a rat subarachnoid hemorrhage (SAH) model. The SAH model was induced by injection of 0.3 ml fresh arterial, non-heparinized blood into the prechiasmatic cistern in 20 s. Anti-AQP-4 antibody, minocycline (an inhibitor of MMP-9), or 2-methoxyestradiol (an inhibitor of HIF-1α), was administered intravenously at 2 and 24 h after SAH. Brain samples were extracted at 48 h after SAH and examined for protein expressions, BBB impairment, and brain edema. Following SAH, remarkable edema and BBB extravasations were observed. Compared with the control group, the SAH animals have significantly upregulated expressions of HIF-1α, AQP-4, and MMP-9, in addition to decreased amounts of laminin and tight junction proteins. Brain edema was repressed after inhibition of AQP-4, MMP-9, or HIF-1α. Although BBB permeability was also ameliorated after inhibition of either HIF-1α or MMP-9, it was not modulated after inhibition of AQP-4. Inhibition of MMP-9 reversed the loss of laminin. Finally, inhibition of HIF-1α significantly suppressed the level of AQP-4 and MMP-9, which could induce the expression of laminin and tight junction proteins. Our results suggest that HIF-1α plays a role in brain edema formation and BBB disruption via a molecular signaling pathway involving AQP-4 and MMP-9. Pharmacological intervention of this pathway in patients with SAH may provide a novel therapeutic strategy for early brain injury.

Melatonin activates the Nrf2-ARE pathway when it protects against early brain injury in a subarachnoid hemorrhage model.

Melatonin has beneficial effects against early brain injury (EBI) by modulating cerebral oxidative stress after experimental subarachnoid hemorrhage (SAH); however, few investigations relate to the precise underlying molecular mechanisms. To date, the relation between melatonin and nuclear factor erythroid 2-related factor 2 and antioxidant responsive element (Nrf2-ARE) pathway has not been studied in SAH models. This study was undertaken to evaluate the influence of melatonin on Nrf2-ARE pathway in rats after SAH. Adult male SD rats were divided into four groups: (i) control group (n=18); (ii) SAH group (n=18); (iii) SAH+vehicle group (n=18); and (iv) SAH+melatonin group (n=18). The rat SAH model was induced by injection of 0.3mL fresh arterial, nonheparinized blood into the prechiasmatic cistern in 20s. In SAH+melatonin group, melatonin was administered i.p. at 150mg/kg at 2 and 24hr after the induction of SAH. Brain samples were extracted at 48hr after SAH. Treatment with melatonin markedly increased the expressions of Nrf2-ARE pathway-related agents, such as Nrf2, heme oxygenase-1, NAD(P)H:quinone oxidoreductase 1, and glutathione S-transferase α-1. Administration of melatonin following SAH significantly ameliorated EBI, including brain edema, blood-brain barrier (BBB) impairment, cortical apoptosis, and neurological deficits. In conclusion, post-SAH melatonin administration may attenuate EBI in this SAH model, possibly through activating Nrf2-ARE pathway and modulating cerebral oxidative stress by inducing antioxidant and detoxifying enzymes.