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Shikonin (SHK) has been evidenced to possess effects against various cancer cells. However, poor aqueous solubility and high toxicity restrict its application. In the study, RGD-decorated liposomes loaded with SHK (RGD-Lipo-SHK) were prepared via thin-film hydration method. Characterization and cellular uptake of liposomes was evaluated. Cytotoxicity of blank liposomes and different SHK formulations was measured against breast cancer cells (MDA-MB-231, MCF-7, and MCF-10A). Anti-tumour effects and pharmacokinetic parameters of different SHK formulations were appraised in tumour spheroids and in rat model, respectively. Liposomes displayed a particle size of less than 127 nm with a polydispersity index about 0.21. The encapsulation efficiency was about 91% for SHK, and drug leakage rate of liposomes was less than 6%. RGD-Lipo-SHK showed superior cellular internalization in the αvß3-positive MDA-MB-231 cells. Blank liposomes had no cytotoxicity to MDA-MB-231 and MCF-7 cells. Howbeit, different SHK formulations obviously inhibited proliferation of MCF-10A cells, especially free SHK. Meanwhile, RGD-Lipo-SHK significantly inhibited growth inhibition of tumour spheroids. The pharmacokinetics study indicated that the peak concentration, area under plasma concentration-time curves, half-life, and mean residence time of RGD-Lipo-SHK distinctly increased compared with those of free SHK. Altogether, these results demonstrated RGD-Lipo-SHK could reduce cytotoxicity, strengthen the antitumor-targeted effect, and prolong circulation time, which provides a foundation for further in vivo experimentations.
Assuntos
Lipossomos , Naftoquinonas , Humanos , Ratos , Animais , Naftoquinonas/farmacologia , Células MCF-7 , Oligopeptídeos , Linhagem Celular TumoralRESUMO
Recently, oxidative stress is strongly associated with lead (Pb)-induced neurotoxicity. We reported previously that Astragaloside IV (AS-IV) possesses potent antioxidant properties. Here, we evaluate the hypothesis that AS-IV attenuates lead acetate (PbAc)-mediated inhibition of neurite outgrowth might mainly result from its antioxidant property via serine/threonine protein kinase (Akt)-dependent activation of the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway. Interestingly, AS-IV attenuates PbAc-induced inhibition of neurite outgrowth and displayed potential antioxidant properties by inhibiting reactive oxygen species (ROS). Concomitantly, AS-IV enhanced phase II detoxifying enzymes such as heme oxygenase 1 (HO-1), thioredoxin reductase (TrxR), and glutamate cysteine ligase catalytic subunit (GCLc). Conversely, AS-IV had no effect on GCL modulatory subunit (GCLm) and superoxide dismutase (SOD) activity/expression. Furthermore, AS-IV evoked Akt phosphorylation, and subsequent induced phosphorylation of glycogen synthase kinase-3ß (GSK-3ß) at Ser9 (that is, inactivation), which stimulated Nrf2-mediated antioxidant response element (ARE)-containing activation. Importantly, Akt locates upstream of GSK-3ß and regulates phase II detoxifying enzymes gene expression through Nrf2 nuclear accumulation in PC12 cells exposed to PbAc. Noteworthy, these results were further confirmed through signalling pathway inhibitors, dominant negative mutant and short hairpin RNA technology. Collectively, these in vitro findings suggest that AS-IV attenuates PbAc-induced inhibition of neurite outgrowth attributed to its antioxidant properties and may be a promising candidate for the treatment of lead developmental neurotoxicity.
Assuntos
Fator 2 Relacionado a NF-E2/metabolismo , Neuritos/metabolismo , Compostos Organometálicos/toxicidade , Proteínas Proto-Oncogênicas c-akt/metabolismo , Saponinas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Triterpenos/farmacologia , Animais , Regulação da Expressão Gênica/efeitos dos fármacos , Proteínas do Tecido Nervoso/biossíntese , Neuritos/patologia , Células PC12 , RatosRESUMO
Objective: Screening and predicting potential targets for gastrodin antioxidant stress based on network pharmacology methods, and exploring the effect of gastrodin on lead acetate induced oxidative stress in PC12 cells through cell experiments. Methods: Through the Pharmaper database Predict the target of action of gastrodin. Through OMIM and GeneCards to collect oxidative stress targets from database, and intersect with drug targets to obtain drug disease intersection targets; Construct a PPI network diagram using the STRING database. Perform GO enrichment analysis and KEGG pathway enrichment analysis on intersection targets through the DAVID platform. Lead acetate (PbAc) exposure was used to establish a lead poisoning cell model, and intracellular ROS levels, ALB, AKT1, and Caspase-3 levels were measured. Results: A total of 288 targets of gastrodin action, 638 targets related to oxidative stress, and 62 drug disease intersection targets were obtained, among which core targets such as ALB, AKT1, CASP3 may be closely related to oxidative stress. KEGG pathway analysis showed that gastrodin antioxidant stress mainly involved in lipid, cancer pathway and other signaling pathways. The results of the cell experiment showed that 50 µM is the optimal effective concentration for PbAc induced ROS production in PC12 cells. Gastrodin significantly increased the ROS content of PC12 cells treated with PbAc, Upregulation of ALB expression and downregulation of AKT1 and CASP3 expression. Conclusions: Gastrodin may alleviate PbAc-induced ROS in PC12 cells, indicating potential protective effects against oxidative stress. Further studies are needed to confirm these findings and explore the underlying mechanisms.
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Hemostats that can strongly adhere to wound tissue and are easy to remove when stopping bleeding are favored for the control of noncompressible hemorrhage. Here, we prepared a citric acid (CA)-crosslinked and N-hydroxysuccinimide (NHS) ester-activated carboxymethyl cellulose (CMC-NHS) aerogel for noncompressible hemostasis. CA was used to crosslink CMC to form a strengthened structure. NHS ester was introduced to activate the adhesion of CMC-NHS aerogel to wound tissue and promoted blood coagulation through the formation of amide crosslinks between CMC and erythrocytes and free blood proteins. The plentiful carboxyl groups could also trigger the intrinsic coagulation pathway. Thus, the aerogel could quickly adhere to wound tissue to stop bleeding, and then could be easily removed when fully hydrated as CMC was dissolved at the adhesion interface. The aerogel also had good biocompatibility and antibacterial capability. Overall, CMC-NHS aerogel is a competitive hemostat for the control of noncompressible hemorrhage.
Assuntos
Adesivos , Carboximetilcelulose Sódica , Humanos , Hemorragia , Coagulação Sanguínea , ÉsteresRESUMO
Prenatal and/or early postnatal exposure to lead (Pb) may be associated with deficits in cognitive function in the toddler offspring, and oxidative stress likely play a central role in mediating these adverse effects. Here, we tested the hypothesis that ameliorative effect of ferulic acid (FA) on lead-induced cognitive deficits attributed to its antioxidant properties in a nuclear factor (erythroid-derived 2)-like 2 (Nrf2)-dependent manner in the context of prenatal Pb exposure. To test this hypothesis, Nrf2 knockout and C57BL/6 wild type mouse dams were exposed/unexposed to PbAc (250 ppm) during gestation day 5 to postnatal day 14 via drinking water, and FA (50 mg/kg)/vehicle was administered orally to dams for 31 d. Spatial learning and memory in pups was assessed by Morris water maze. Biochemical assays, real-time PCR, western blot techniques were employed to evaluate oxidative stress and signaling pathways in the brain of pups. We report that lead acetate (PbAc) leads to deficits in cognitive functions in offspring, which were partially attenuated by FA (P<.05). In parallel, pretreatment with FA also significantly inhibited the PbAc-induced oxidative stress, as indicated by a change in NAD+/NADH ratio, glutathione (GSH) and malondialdehyde contents (all P<.05). Interestingly, FA significantly elevated the glutamate cysteine ligase and heme oxygenase 1 at levels of transcription and translation in both mice exposed and unexposed to Pb, increasing de novo synthesis of GSH (all P<.05). Furthermore, maternal FA administration activates extracellular signal-regulated kinases 1 and 2 and promotes more Nrf2 nuclear accumulation by increasing the Nrf2 total protein in brain of offspring mice (all P<.05). Conversely, FA failed to influence Pb-induced both memory deficits and oxidative stress in offspring of Nrf2 knockout mice (all P≥.05), suggesting that Nrf2 is essential in mediating the cognition-enhancing and antioxidant effects of FA. Overall, our results demonstrate that FA protects against Pb-induced offspring's cognitive deficits, suggesting that it is a promising candidate for the treatment of Pb toxicity.
Assuntos
Disfunção Cognitiva/prevenção & controle , Ácidos Cumáricos/uso terapêutico , Chumbo/efeitos adversos , Fator 2 Relacionado a NF-E2/metabolismo , Efeitos Tardios da Exposição Pré-Natal/prevenção & controle , Substâncias Protetoras/uso terapêutico , Animais , Animais Recém-Nascidos , Disfunção Cognitiva/induzido quimicamente , Disfunção Cognitiva/metabolismo , Feminino , Camundongos Endogâmicos C57BL , Gravidez , Efeitos Tardios da Exposição Pré-Natal/induzido quimicamente , Efeitos Tardios da Exposição Pré-Natal/metabolismoRESUMO
In this study, based on the oxidative decolorization of three azo dyes (Orange G (OG), Acid Orange 7 (AO7) and Reactive Black 5 (RB5)) with hydroxyl radicals generated in Fenton system, we have successfully established three types of azo dyes spectrophotometric methods for measuring aqueous hydrogen peroxide (H2O2) concentration. The decolorization extent of OG, AO7 and RB5 at the corresponding characteristic wavelengths of 478â¯nm, 484â¯nm and 597â¯nm are proportion to the concentration of H2O2 in aqueous solutions. Under the selected reaction conditions, three well linear correlations between the depletion of azo dyes and the H2O2 concentration are established in the range of 0.45-175⯵molâ¯L-1 of OG, 0.36-120⯵molâ¯L-1 of AO7 and 0.44-175⯵molâ¯L-1 of RB5, respectively. These proposed spectrophotometric methods are enough accurate to measure low concentrations of H2O2 in practical water samples and monitor the variations of H2O2 concentration during the phenol degradation in the Cu(II)/HCO3-/H2O2 process.