RESUMEN
The present study aimed to explore the potential mechanism of the effect of hyperbaric oxygenation (HBO) preconditioning on cerebral ischemia and reperfusion injury (CIRI). GSE23160 dataset was used to identify differentially expressed genes (DEGs) from striatum between the middle cerebral artery occlusion (MCAO)/reperfusion and sham rats. The gene clusters with continuous increase and decrease were identified by soft clustering analysis in Mfuzz, and functional enrichment analysis of these genes was performed using clusterProfiler package. The intersection set of the genes with significantly altered expression at post-reperfusion 2, 8, and 24 h were screened in comparison to 0 h (sham group), and the expression of these genes was detected in the MCAO/reperfusion model and HBO preconditioning groups by real-time PCR (RT-PCR) and western blotting. A total of 41 upregulated DEGs, and 7 downregulated DEGs were detected, among which the expression of Gpr84 and Ggta1 was significantly upregulated at each reperfusion phase as compared to the sham group, while the expression of Kcnk3 was significantly downregulated except in the postreperfusion 8 h in the striatum group. RT-PCR and western blotting analyses showed that the expression of Ggta1, Gpr84, and Kcnk3 genes between the MCAO/reperfusion and sham rats were consistent with the bioinformatics analysis. In addition, the HBO preconditioning reduced the expression of Ggta1 and Gpr84 and increased the expression of Kcnk3 in MCAO/reperfusion rats. Kcnk3, Ggta1, and Gpr84 may play a major role in HBO-mediated protection of the brain against CIRI.
Asunto(s)
Isquemia Encefálica , Oxigenoterapia Hiperbárica , Daño por Reperfusión , Animales , Infarto de la Arteria Cerebral Media , Ratas , Ratas Sprague-Dawley , Daño por Reperfusión/genética , Daño por Reperfusión/prevención & controlRESUMEN
Ischemic stroke is one of the leading causes of mortality and disability worldwide. Previous studies have indicated that hyperbaric oxygen preconditioning (HBO-PC) can induce neuroprotection against focal cerebral ischemia. However, the underlying mechanisms are still not fully understood, and the optimal regimen for preconditioning must be confirmed. In the present study, we designed eight preconditioning regimens and compared their neuroprotective effects. Hyperbaric oxygen preconditioning every other day for there sessions exhibited the best neuroprotective effect; the infarct volume was reduced by almost 50% at 48 h after middle cerebral artery occlusion. We also found that HBO-PC significantly increased the microvessel density and the CD31-positive cells in the penumbra at 72 h after stroke. These results indicate that angiogenesis is involved in the neuroprotection induced by HBO-PC. Moreover, we explored the roles of HIF-1α and angiogenic factors in the angiogenesis process induced by HBO-PC. The results from western blotting demonstrated that protein expression of Ang-2 in the HBO-PC group was significantly increased. In conclusion, HBO-PC reduced brain injury and improved neurological function after focal cerebral ischemia, as partly mediated by the increased microvessel density in the penumbra, and this effect may result from the upregulation of Ang-2.
Asunto(s)
Isquemia Encefálica/prevención & control , Oxigenoterapia Hiperbárica/métodos , Precondicionamiento Isquémico , Neovascularización Fisiológica/fisiología , Neuroprotección/fisiología , Animales , Modelos Animales de Enfermedad , Regulación de la Expresión Génica , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Masculino , Examen Neurológico , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/metabolismo , Ratas , Ratas Sprague-Dawley , Factores de Tiempo , Factor de von Willebrand/metabolismoRESUMEN
The major objective was to explore the effect of early hyperbaric oxygen (HBO) therapy on the tissue structure, apoptosis, and metalloproteinases of kidney cells in Goto-Kakizaki (GK) rats with type 2 diabetes mellitus. GK rats (n = 24) were divided randomly and evenly into model, metformin hydrochloride (MH), and hyperbaric oxygen (HBO) groups, while healthy Wistar rats (n = 8) were used as normal control group. The healthy rats in the normal control group and the GK rats in the model group were both intragastrically administered with purified water (5 mL/kg) once per day. Meanwhile, the rats in the MH group received intragastric administration of MH (250 mg/kg) once daily, while the rats in the HBO group inhaled pure oxygen under a constant pressure (0.15 MPa) for 30 min. After 3 weeks of treatment, the body weight of each rat was measured, and the blood samples were collected from tails. Subsequently, the kidneys of all rats were excised for weighing mass and further examination. For each renal sample, the sections were firstly embedded with paraffin and sliced to prepare histopathologic sections stained using HE, PAS and Masson, respectively, for subsequent observation with optical microscopy. Later, the apoptosis of kidney cells was examined using the TUNEL method by computing the apoptotic index. Furthermore, the histopathologic sections were also examined using the immunohistochemistry approach with Caspase-3, MMP-2, and TIMP-2 antibodies, respectively. At the same time, the plasma concentration of TGF-ß1 of the rats in each group was detected using ELISA method. These resultant data showed that the pathological changes of the HBO group were less than those of the model group with respect to increased glomerular volume density of mesangial cells, broadening mesangial matrix and thickening basement membrane as well as swelling renal tubular epithelial cells. The index of cell apoptosis and Caspase-3 expression in the HBO group showed no significant differences (P > 0.05) compared with those in the normal control and MH groups respectively, but demonstrated significant decrease compared with that in the model group (P < 0.01). Meanwhile, the MMP-2 and TIMP-2 expressions of the HBO group were stronger than those in the model and MH groups, but weaker than those in the normal control group (P < 0.05). Although the plasma concentration of TGF-ß1 in HBO, MH and model groups was greater than that in the normal control group, no significant statistical difference was distinguished among these four groups (P > 0.05). These results indicate that the HBO treatment can inhibit the apoptosis and Caspase-3 expression of renal cells of GK rats, adjust the activity of MMP-2 and its inhibitors, and reduce the accumulation of extracellular matrix. This implies that the HBO treatment might protect renal tissues, thus delaying occurrence and retaining development of diabetic nephropathy.
Asunto(s)
Apoptosis , Diabetes Mellitus Experimental/fisiopatología , Oxigenoterapia Hiperbárica , Riñón/citología , Animales , Caspasa 3/metabolismo , Diabetes Mellitus Tipo 2/fisiopatología , Nefropatías Diabéticas/terapia , Metaloproteinasa 2 de la Matriz/metabolismo , Oxígeno/administración & dosificación , Ratas , Ratas Wistar , Inhibidor Tisular de Metaloproteinasa-2/metabolismo , Factor de Crecimiento Transformador beta1RESUMEN
AIMS: To evaluate the effects of hyperbaric oxygen (HBO) therapy on MMP-2 and MMP-9 expression and spinal cord edema after acute spinal cord injury (SCI). MAIN METHODS: Ninety-six healthy adult Sprague − Dawley rats were randomly divided into 4 groups: sham, sham + HBO, SCI, and SCI + HBO. Each group was subdivided into 4 subgroups of 6 rats each: 24 h, 48 h, 72 h, and 5 d post-injury. The expression levels of matrix metalloproteinase-2 (MMP-2), MMP-9, interleukin (IL-6), and vascular endothelial growth factor (VEGF) were measured using ELISA. The water content of the spinal cord was also measured by Elliot equation. KEY FINDING: MMP-2 and MMP-9 levels and spinal cord water content increased significantly in the SCI group compared to those in the sham and sham + HBO groups at different time points after SCI (P < 0.01). The MMP-2 level was significantly different between the SCI and SCI + HBO groups at 72 h after SCI (P < 0.05), while the MMP-9 level was significantly different at 48 h, 72 h, and 5 d (P < 0.05). The water content of the spinal cord was significantly higher in the SCI group at 5 d after SCI (P < 0. 01). MMP-2 (P < 0.05) and MMP-9 (P < 0.01) levels were positively correlated with spinal cord water content. SIGNIFICANCE: HBO reduced SCI-induced spinal cord edema, stabilized the blood-spinal cord barrier, and promoted recovery of neuronal function by down regulating the expression of IL-6, MMP-2, and MMP-9 and up regulating the expression of VEGF.