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1.
Acta Pharmaceutica Sinica B ; (6): 2330-2347, 2022.
Article in English | WPRIM | ID: wpr-929377

ABSTRACT

Ischemic brain stroke is pathologically characterized by tissue acidosis, sustained calcium entry and progressive cell death. Previous studies focusing on antagonizing N-methyl-d-aspartate (NMDA) receptors have failed to translate any clinical benefits, suggesting a non-NMDA mechanism involved in the sustained injury after stroke. Here, we report that inhibition of intracellular proton-sensitive Ca2+-permeable transient receptor potential vanilloid 3 (TRPV3) channel protects against cerebral ischemia/reperfusion (I/R) injury. TRPV3 expression is upregulated in mice subjected to cerebral I/R injury. Silencing of TRPV3 reduces intrinsic neuronal excitability, excitatory synaptic transmissions, and also attenuates cerebral I/R injury in mouse model of transient middle cerebral artery occlusion (tMCAO). Conversely, overexpressing or re-expressing TRPV3 increases neuronal excitability, excitatory synaptic transmissions and aggravates cerebral I/R injury. Furthermore, specific inhibition of TRPV3 by natural forsythoside B decreases neural excitability and attenuates cerebral I/R injury. Taken together, our findings for the first time reveal a causative role of neuronal TRPV3 channel in progressive cell death after stroke, and blocking overactive TRPV3 channel may provide therapeutic potential for ischemic brain injury.

2.
Article in Chinese | WPRIM | ID: wpr-928023

ABSTRACT

This study aims to explore the pharmacodynamic effect of baicalin on rat brain edema induced by cerebral ischemia reperfusion injury and discuss the mechanism from the perspective of inhibiting astrocyte swelling, which is expected to serve as a refe-rence for the treatment of cerebral ischemia with Chinese medicine. To be specific, middle cerebral artery occlusion(suture method) was used to induce cerebral ischemia in rats. Rats were randomized into normal group, model group, high-dose baicalin(20 mg·kg~(-1)) group, and low-dose baicalin(10 mg·kg~(-1)) group. The neurobehavior, brain index, brain water content, and cerebral infarction area of rats were measured 6 h and 24 h after cerebral ischemia. Brain slices were stained with hematoxylin and eosin(HE) for the observation of pathological morphology of cerebral cortex after baicalin treatment. Enzyme-linked immunosorbent assay(ELISA) was employed to determine the content of total L-glutathione(GSH) and glutamic acid(Glu) in brain tissue, Western blot to measure the content of glial fibrillary acidic protein(GFAP), aquaporin-4(AQP4), and transient receptor potential vanilloid type 4(TRPV4), and immunohistochemical staining to observe the expression of GFAP. The low-dose baicalin was used for exploring the mechanism. The experimental results showed that the neurobehavioral scores(6 h and 24 h of cerebral ischemia), brain water content, and cerebral infarction area of the model group were increased, and both high-dose and low-dose baicalin can lower the above three indexes. The content of GSH dropped but the content of Glu raised in brain tissue of rats in the model group. Low-dose baicalin can elevate the content of GSH and lower the content of Glu. According to the immunohistochemical staining result, the model group demonstrated the increase in GFAP expression, and swelling and proliferation of astrocytes, and the low-dose baicalin can significantly improve this situation. The results of Western blot showed that the expression of GFAP, TRPV4, and AQP4 in the cerebral cortex of the model group increased, and the low-dose baicalin reduce their expression. The cerebral cortex of rats in the model group was severely damaged, and the low-dose baicalin can significantly alleviate the damage. The above results indicate that baicalin can effectively relieve the brain edema caused by cerebral ischemia reperfusion injury in rats, possibly by suppressing astrocyte swelling and TRPV4 and AQP4.


Subject(s)
Animals , Aquaporin 4/genetics , Astrocytes , Brain Edema/drug therapy , Brain Ischemia/metabolism , Flavonoids , Infarction, Middle Cerebral Artery/drug therapy , Rats , Rats, Sprague-Dawley , Reperfusion , TRPV Cation Channels/therapeutic use
3.
Article in Chinese | WPRIM | ID: wpr-927892

ABSTRACT

Objective: To investigate the effects of Zhongfeng capsule on the autophagy-related proteins expression in rats with cerebral ischemia/reperfusion injury (CI/ RI), and to explore its neural protection mechanisms of the decoction. Methods: Rat middle cerebral artery ischemia/reperfusion injury model (ischemia for 2 h, reperfusion for 24 h) was prepared by the improved line plug method. Sixty male SD rats were randomly divided into sham operation group, model group, butylphthalide group(0.054 g/kg), Zhongfeng capsule high-dose groups (1.08 g/kg), Zhongfeng capsule middle-dose groups (0.54 g/kg), Zhongfeng capsule low-dose groups (0.27 g/kg), with 10 rats in each group. Rats were treated with Zhongfeng capsule by gavage once a day for 10 days. The rats were sacrificed and the brain tissue was obtained after the experiment in each group. Score neurological deficit was evaluated after 24 h of the last intervention in rat of each group. The pathological changes of brain tissue were observed by HE staining. The serum levels of estradiol (E2) and follicle stimulating hormone (FSH) were determined by ELISA. The expressions of key genes and proteins of PI3K/Akt/Beclin1 signaling pathway in brain tissue were detected by qRT-PCR and Western blot respectively. Results: Compared with the sham operation group, the body weight and protein expressions of p-PI3k and p-Akt in brain tissue of rats were decreased significantly in the model group, while the brain index, neurological deficit score, gene and protein expressions of Beclin1 and LC3 were increased markedly in the model group(P<0.05 or P<0.01). In the model group, nerve cells of brain tissue were loosely packed, interstitial edema, triangular in shape, nuclear pyknosis and dark-blue staining were observed. Compared with the model group, the body weight of rats was increased obviously, the neurological deficit score was decreased significantly and the pathological injury of brain tissue was alleviated evidently in high-dose of Zhongfeng capsule group (P<0.05). The brain index, the gene and protein expressions of Beclin1 and LC3 were decreased apparently in Zhongfeng capsule treatment groups(P<0.05 or P<0.01), while the expressions of p-PI3k and p-Akt in brain tissue were increased evidently in Zhongfeng capsule treatment groups(P<0.05 or P<0.01). Conclusion: Zhongfeng capsule can inhibit autophagy and improve brain neurons lesion of CIRI rats, the mechanism may be related to regulate the expression of Beclin1 and LC3 in PI3K/Akt/Beclin1 signaling pathway.


Subject(s)
Animals , Autophagy-Related Proteins/pharmacology , Beclin-1/metabolism , Body Weight , Brain , Brain Ischemia/metabolism , Male , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Rats , Rats, Sprague-Dawley , Reperfusion Injury/drug therapy
4.
Braz. j. med. biol. res ; 55: e12145, 2022. graf
Article in English | LILACS-Express | LILACS | ID: biblio-1384152

ABSTRACT

Dexmedetomidine (DEX) is known to provide neuroprotection against cerebral ischemia and reperfusion injury (CIRI), but the exact mechanisms remain unclear. This study was conducted to investigate whether DEX pretreatment conferred neuroprotection against CIRI by inhibiting neuroinflammation through the JAK2/STAT3 signaling pathway. Middle cerebral artery occlusion (MCAO) was performed to establish a cerebral ischemia/reperfusion (I/R) model. Specific-pathogen-free male Sprague-Dawley rats were randomly divided into Sham, I/R, DEX, DEX+IL-6, and AG490 (a selective inhibitor of JAK2) groups. The Longa score, TTC staining, and HE staining were used to evaluate brain damage. ELISA was used to exam levels of TNF-α. Western blotting was used to assess the levels of JAK2, phosphorylated-JAK2 (p-JAK2), STAT3, and phosphorylated-STAT3 (p-STAT3). Our results suggested that both pretreatment with DEX and AG490 decreased the Longa score and cerebral infarct areas following cerebral I/R. After treatment with IL-6, the effects of DEX on abrogating these pathological changes were reduced. HE staining revealed that I/R-induced neuronal pathological changes were attenuated by DEX application, consistent with the AG490 group. However, these effects of DEX were abolished by IL-6. Furthermore, TNF-α levels were significantly increased in the I/R group, accompanied by an increase in the levels of the p-JAK2 and p-STAT3. DEX and AG490 pretreatment down-regulated the expressions of TNF-α, p-JAK2, and p-STAT3. In contrast, the down-regulation of TNF-α, p-JAK2, and p-STAT3 induced by DEX was reversed by IL-6. Collectively, our results indicated that DEX pretreatment conferred neuroprotection against CIRI by inhibiting neuroinflammation via negatively regulating the JAK2/STAT3 signaling pathway.

5.
Medisan ; 25(5)2021. ilus
Article in Spanish | LILACS, CUMED | ID: biblio-1346548

ABSTRACT

Los accidentes cerebrovasculares se han mantenido, a nivel mundial, como la tercera causa de muerte y la primera de discapacidad. Para disminuir la incidencia de casos de isquemia o hemorragia cerebral, así como sus consecuencias, se deben poseer los conocimientos sobre dichas entidades clínicas, los factores de riesgo asociados y las alternativas preventivas y terapéuticas como estrategias neuroprotectoras. Muchas de las intervenciones médicas realizadas hasta la fecha en modelos animales han resultado insatisfactorias en la fase clínica. Por ello, se realizó una revisión de las publicaciones más recientes donde se abordan los modelos experimentales para la isquemia cerebral más utilizados en las evaluaciones de las terapias neuroprotectoras, y se pudo concluir que si se analizan los protocolos empleados en la fase preclínica podrán optimizarse las investigaciones para lograr resultados más acertados en este campo.


The strokes have been considered, worldwide, as the third cause of death and the first cause of disability. To diminish the incidence of ischemia cases or cerebral hemorrhage, as well as their consequences, one should have the knowledge on this clinical entities, the associated risk factors and preventive and therapeutic alternatives as neuroprotector strategies. Many of the medical interventions carried out so far in animal models have been unsatisfactory in the clinical phase. Reason why, a review of the most recent publications was carried out, where the most used experimental models for the cerebral ischemia in the evaluations of the neuroprotector therapies are approached, and it was concluded that if protocols used in the preclinic phase are analyzed, the investigations could be optimize to achieve more relevant results in this field.


Subject(s)
Brain Ischemia , Neuroprotective Agents , Stroke , Risk Factors , Models, Animal
6.
Int. j. morphol ; 39(3)jun. 2021.
Article in English | LILACS-Express | LILACS | ID: biblio-1385408

ABSTRACT

SUMMARY: Cerebral ischemia has not only a high mortality rate, which is the second leading cause of death worldwide, but is also responsible for severe disabilities in working age individuals, generating enormous public expending for treatment and rehabilitation of the affected individuals. The role of microRNAs in the pathophysiology of cerebral ischemia has been highlighted in current investigations. In addition, recent studies have also highlighted physical exercise as a possible protective factor both in the prevention and in the effects of cerebral ischemia, placing it as an important study resource. Thus, we investigated the role of physical exercise in experimental cerebral ischemia associated with the expression of microRNA-27b. 16 animals were used, divided into four experimental groups: Control, Physical Exercise, Cerebral Ischemia and Cerebral Ischemia associated with Physical Exercise. The real-time PCR methodology was used to analyze the expression of microRNA-27b. Although there were no statistically significant differences in the expression of microRNA-27b between the groups studied, the increased expression of microRNA-27b in the Physical Exercise group indicates its neuroprotective role in the pathophysiology of cerebral ischemia.


RESUMEN: La isquemia cerebral no solo tiene una alta tasa de mortalidad y es la segunda causa principal de muerte en todo el mundo, sino también es la causa de enfermedades invalidantes en personas en edad laboral, lo que genera un gasto público enorme para el tratamiento y la rehabilitación de las personas afectadas. El papel de los microARN en la fisiopatología de la isquemia cerebral se ha destacado en las investigaciones actuales. Además, estudios recientes también han destacado el ejercicio físico como un posible factor protector tanto en la prevención como en los efectos de la isquemia cerebral, situándolo como un importante recurso de estudio. Por lo tanto, investigamos el papel del ejercicio físico en la isquemia cerebral experimental asociada con la expresión del microARN-27b. Se utilizaron 16 animales, divididos en cuatro grupos experimentales: Control, Ejercicio Físico, Isquemia Cerebral e Isquemia Cerebral asociada al Ejercicio Físico. Se utili- zó la metodología de PCR en tiempo real para analizar la expresión de microARN-27b. Aunque no se observaron diferencias estadísticamente significativas en la expresión de microARN-27b entre los grupos estudiados, la mayor expresión de microARN-27b en el grupo de Ejercicio Físico indica su papel neuroprotector en la fisiopatología de la isquemia cerebral.

7.
Article in Chinese | WPRIM | ID: wpr-876147

ABSTRACT

@#To investigate the neuroprotective effect and possible mechanism of masitinib on cerebral ischemia-reperfusion injury in rats, healthy adult male Sprague-Dawley rats were divided into sham group (n = 12), model group (n = 12), masitinib low dosage group (n = 12), masitinib middle dosage group (n = 12), and masitinib high dosage group (n = 12). All rats was subjected to middle cerebral artery occlusion (MCAO) for two hours and reperfusion except sham group, and received treatment twice per day for 7 days once reperfusion started.Neurological score, infarct volume, and brain water content were detected; some autophagic markers, apoptotic and inflammatory cytokines were evaluated by Western blot and PCR after 7 d of reperfusion. Treatment with masitinib significantly ameliorated neurologic deficit, infarct volume and brain water after I/R injury. Masitinib also decreased the ratio of LC3II/I and the expression of Beclin-1 and increased the expression of p62 in the brain tissues of rats with I/R injury.Furthermore, it could inhibit apoptosis-related proteins and NF-κB expression. Masitinib could relieve the cerebral ischemia-reperfusion injury in rats through inhibiting autophagy and apoptosis.

8.
Organ Transplantation ; (6): 428-2021.
Article in Chinese | WPRIM | ID: wpr-881527

ABSTRACT

Objective To investigate whether Danhong injection can enhance the therapeutic effect of neural stem cell (NSC) transplantation in repairing cerebral ischemia injury by regulating the nuclear factor E2-related factor 2 (Nrf2) signaling pathway. Methods Forty male SD rats were randomly divided into the NSC transplantation group (NSC group), Danhong injection group (DH group), NSC+ Danhong injection group (N+D group), NSC+ Danhong injection group +ML385 group(N+D+M group) and PBS control group (PBS group), 8 rats in each group. All rat models of cerebral ischemia were established by embolization of the middle cerebral artery. Reperfusion was performed at 1.5 h after embolization. All rats in each group received corresponding interventions at 3 d after reperfusion. The neurological function score was evaluated before and 1, 2, 4 weeks after NSC transplantation. All rats were sacrificed at 4 weeks after NSC transplantation. The parameters related to oxidative stress were detected. The expression levels of neuron-specific nuclear protein (NeuN) and von Willebrand factor (vWF) were determined by immunofluorescence staining. Results Before NSC transplantation, the neurological function scores did not significantly differ among different groups (all P > 0.05). At postoperative 1, 2 and 4 weeks, the neurological function scores in the NSC, DH and N+D groups were significantly lower than those in the PBS and N+D+M groups (all P < 0.05). Compared with the PBS and N+D+M groups, the malondialdehyde (MDA) levels were significantly decreased, whereas the superoxide dismutase (SOD) and glutathione peroxidase (GPX) levels were considerably increased in the NSC, DH and N+D groups (all P < 0.05). The GPX level in the N+D+M group was significantly lower than that in the PBS group (P < 0.05). Immunofluorescence staining showed that the transplant NSC in the rat brain migrated to the surrounding area of cerebral infarction and survived, and expressed neuronal marker NeuN and neovascularization marker vWF. However, the number of living NSC in the N+D+M group was significantly lower compared with those in the remaining groups. Conclusions Danhong injection may improve the microenvironment of stem cell transplantation, enhance the survival rate of transplant NSC and improve the therapeutic effect of NSC transplantation for cerebral ischemia injury probably by regulating the Nrf2 signaling pathway.

9.
Article in Chinese | WPRIM | ID: wpr-881390

ABSTRACT

@#PLZ-NPs (PNS-lipid-zein nanoparticles) prepared by co-assembly of Panax notoginseng saponins, lecithin, β-sitosterol and zein were applied for in vitro cell experiment and oral gavage to study the protective effect of cerebral ischemia-reperfusion rats.PLZ-NPs were characterized by Malvin-particle size analyzer and transmission electron microscope (TEM), respectively. The toxicity of PLZ-NPs and free carrier were evaluated by MTT, and the uptake of nanoparticles in Caco-2 cells was analyzed by laser confocal and flow cytometry. The cerebral ischemia reperfusion rat model was established by MCAO method and then be given samples by gavage for 3 days. The brain tissues were taken to stain by 2, 3, 5-triphenyltetrazole chloride (TTC) and the biochemical indicators of MDA, inflammatory cytokines IL-1β and TNF-α, apoptosis-related proteins Bax and Bcl-2 from the harvested brain tissues were detected to evaluate the protective effect of PNS in PLZ-NPs on cerebral ischemia reperfusion. The particle size, PDI, and zeta potential of formed PLZ-NPs were (116.4 ± 0.81) nm, 0.048 and -(31.5 ± 0.31) mV, respectively. The results of MTT showed that the zein lipoprotein carrier was non-toxic to Caco-2 cells. The results of laser confocal and flow cytometry showed that FITC uptake of nanoparticles could be significantly improved in Caco-2 cells.The uptake from the nanoparticles at 4h was 1.76 times of that of the free FITC group.Compared with the model group, the TTC staining images of free drug PNS group and PLZ-NPs group showed certain reduction in the white infarct area.The contents of MDA, IL-1β, TNF-α and Bax were significantly decreased, while the content of Bcl-2 was significantly increased. Furthermore, all parameters of PLZ-NPs group showed better results than those of PNS group, and there was a significant difference (P < 0.05). All results indicated that the prepared PLZ-NPs had good stability and biological safety, and could significantly increase the uptake in intestinal epithelial cells, and effectively protect against the damage caused by cerebral ischemia reperfusion in rats.

10.
Article in Chinese | WPRIM | ID: wpr-907672

ABSTRACT

Objective:To determinate the active ingredients and predicte action targets in Sanhua Decoction for treating cerebral ischemia based on HPLC and network pharmacology methods. Methods:The HPLC analysis was performed on HC-C18 (250 mm × 4.6 mm, 5 μm) with the mobile phase of methanolacetonitrile-0.05% phosphoric acid with gradient elution at the flow rate of 1.0 ml/min and 0.8 ml/min. The detection wavelength was set at 254 nm, the column temperature was at 30 ℃, and the injection volume was 10 μl. TCMIP v2.0 platform was used to search the action targets of rhein, emodin, chrysophanol, hesperidin, magnolol and notopterygiol. We used Cerebral ischemia as a keyword searching the databases such as Genecards, OMIM, TTD, and Disgenet to screen six potential targets for the treatment of cerebral ischemia by active ingredients, construct a protein interaction (PPI) network and a disease-component-target-pathway integration network.Results:The literature search determined that rhein, emodin, chrysophanol, hesperidin, magnolol and qianhuol were the index components for the determination of Sanhua Decoction. Their linear ranges were 0.080 4-0.804 0 μg, 0.015 3-0.382 0 μg, 0.041 8-0.626 4 μg, 0.312 6-3.908 0 μg, 0.037 9-0.568 8 μg, 0.045 3-1.359 6 μg, respectively. The correlation coefficient R 2 is greater than 0.999 0. The content ranges of the above six components in seven samples were 0.887-0.971 mg/g, 0.094-0.101 mg/g, 0.110-0.119 mg/g, 1.494-1.669 mg/g, 0.126-0.145 mg/g and 0.153-0.167 mg/g, respectively. Network pharmacology analysis found that the targets of the six components for the treatment of cerebral ischemia may be TNF, TP53, MAPK14, JUN, IL1B, MYC, ESR1, ICAM1, PTGS2, PPARG and so on. Conclusions:A quality control method forthe six active ingredients in Sanhua Decoction treating cerebral ischemia was established. This method is simple and repeatable. The ten potential targets of the six active ingredients in Sanhua Decoction for the treatment of cerebral ischemia have been clarified, laying a foundation for further research on the action mechanism.

11.
Article in Chinese | WPRIM | ID: wpr-906339

ABSTRACT

Objective:Combined with high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (HPLC-Q-TOF-MS/MS) and network pharmacology, to predict the target and potential mechanism of Gardeniae Fructus in the treatment of cerebral ischemia. Method:HPLC-Q-TOF-MS/MS was used to identify the chemical constituents of Gardeniae Fructus according to the retention time, relative molecular weight, secondary mass spectrometry fragmentation and other information of chromatographic peaks, and combined with literature data. The targets of main active ingredients in Gardeniae Fructus were predicted by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and SwissTargetPrediction database. The potential targets of Gardeniae Fructus against cerebral ischemia were obtained through Online Mendelian Inheritance in Man (OMIM), GeneCards and Kyoto Encyclopedia of Genes and Genomes (KEGG). Gene ontology (GO) function enrichment and KEGG pathway analysis of potential targets were analyzed with the DAVID 6.8. Cytoscape 3.6.0 software was used to construct the network of active components-targets-pathways. At last, Discovery Studio 2016 software was applied in the molecular docking verification between the key active ingredients and potential protein targets. Result:A total of 40 chemical constituents in Gardeniae Fructus were identified, including iridoids, diterpenoid pigments, organic acids, monoterpenoids and other components. According to the main active ingredients, 208 potential targets were predicted, 560 disease targets related to cerebral ischemia were retrieved, 59 key targets were selected by mapping component targets with disease targets. These targets could act on key target proteins such as tumor necrosis factor (TNF), Caspase-3 (CASP3) and CASP8, and participate in the regulation of TNF, phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt), hypoxia inducible factor 1 (HIF-1) and other signal pathways. Molecular docking results showed that geniposide could interact with targets of prostaglandin G/H synthase 2 (PTGS2), TNF<italic> </italic>and nuclear transcription factor-<italic>κ</italic>B p65 (RELA), crocin Ⅰ could interact with interleukin-2 (IL-2). Conclusion:Geniposide, crocin Ⅰ and other ingredients in Gardeniae Fructus can play a role of anti-inflammatory and inhibiting apoptosis to prevent or treat cerebral ischemic diseases by up-regulating protein expression level of RELA and IL-2, down-regulating protein expression level of TNF, CASP8, CASP3 and matrix metalloproteinase 2 (MMP2), and regulating TNF, PI3K/Akt and HIF-1 signaling pathways.

12.
Article in Chinese | WPRIM | ID: wpr-906206

ABSTRACT

Objective:To investigate the effect of rhein on aquaporin 4 (AQP4) and brain edema after cerebral ischemia and the role of microglia-mediated inflammation in this process. Method:The modified thread embolization method was selected to establish the cerebral ischemia model of the right middle cerebral artery embolism (MCAO) in rats. The rats were divided into sham operation group, model group, minocycline group, and high, medium and low-dose rhein groups (3.46,1.73,0.865 mg·kg<sup>-1</sup>). The neurobehavioral function was measured by a modified neurobehavioral score. Wet and dry weight methods were used to measure the changes of water content in brain tissue of rats with cerebral ischemic injury. Western blot was used to detect the expressions of interferon-<italic>γ</italic> (IFN-<italic>γ</italic>) and interleukin-2 (IL-2) in the peripheral ischemic area of rats in each group. Immunofluorescence double labeling method was used to detect the expressions and localization of microglia fine markers Iba-1 and AQP4. Result:Compared with the sham operation group, neurological function score and water content on the side of brain tissue injury of the model group were significantly increased (<italic>P</italic><0.05). Compared with the model group, the neurological function score and the water content of the brain tissue of each drug group were reduced (<italic>P</italic><0.05, <italic>P</italic><0.01). Compared with sham operation group, the protein expressions of IFN-<italic>γ</italic> and IL-2 in the model group increased significantly (<italic>P</italic><0.05). Compared with the model group, the protein expressions of IFN-<italic>γ</italic> and IL-2 in the peripheral area of cerebral ischemia of each drug group were significantly improved (<italic>P</italic><0.05, <italic>P</italic><0.01). Immunofluorescence double staining results showed that compared with the sham operation group, the model group showed significant increase in the fluorescence expression of AQP4 protein on activated microglia, while each drug group could reduce the fluorescence expression of AQP4 protein on activated microglia, different levels of activated microglia markers Iba-1 and AQP4 were co-localized in the peripheral area of cerebral ischemia in each group. Conclusion:Rhein could reduce the degree of brain edema caused by cerebral ischemic injury, and its mechanism may be related to the inhibition of microglia-mediated neuroinflammation and the down-regulation of AQP4 expression.

13.
Article in Chinese | WPRIM | ID: wpr-906016

ABSTRACT

Objective:To explore the effects of Huatan Tongluo Decoction (HTTLD) on the morphology and function of brain tissues and intestine in rats with cerebral ischemia/reperfusion based on the gut-brain axis. Method:Sixty SPF male rats were randomly divided into a sham operation group, a model group, high- (28.66 g·kg<sup>-1</sup>), medium- (14.33 g·kg<sup>-1</sup>), and low-dose (7.16 g·kg<sup>-1</sup>) HTTLD groups, and an edaravone (4 g·kg<sup>-1</sup>)+<italic>Clostridium butyricum</italic> (5.0×10<sup>8</sup> cfu·mL<sup>-1</sup>) group. The model was established by focal cerebral ischemia/reperfusion in rats. The drugs were administered by gavage. The brain tissue injury was determined by neurological deficit score and 2,3,5-triphenyl tetrazolium chloride (TTC) staining. The effect of cerebral ischemia/reperfusion on intestinal motility was assessed by the propulsion rate of small intestine. The intestinal mucosal cell damage was evaluated by the pathomorphological examination of the duodenal mucosa. Enzyme-linked immunosorbent assay (ELISA) was used to determine the content of <italic>D</italic>-lactate (<italic>D</italic>-LAC), diamine oxidase (DAO), and bacterial endotoxin (lipopolysaccharide, LPS) in serum. Western blot was used to detect the expression of Occludin, Claudin-5, and zonula occludens 1 (ZO-1) in the duodenum. Result:After cerebral ischemia/reperfusion, rats developed neurological deficit symptoms. The neurological deficit score in the model group was higher than that in the sham operation group (<italic>P<</italic>0.01). Compared with the model group, the high- and medium-dose HTTLD groups could relieve the symptoms of neurological deficits and lower neurological deficit scores (<italic>P<</italic>0.01). The results of TTC staining showed that the model group presented obvious infarcts in brain tissues compared with the sham operation group (<italic>P<</italic>0.01). The cerebral infarction volumes of HTTLD groups were reduced compared with that in the model group (<italic>P<</italic>0.01), especially the high-dose HTTLD group, and the effect was dose-dependent. Furthermore, the propulsion rate of small intestine in the model group was significantly reduced compared with that in the sham operation group (<italic>P<</italic>0.01). Compared with the model group, HTTLD groups could increase propulsion rates of small intestine (<italic>P<</italic>0.01), especially the high-dose HTTLD group, and the effect was dose-dependent. After cerebral ischemia/reperfusion, obvious duodenal mucosal damage could be observed, which was relieved after the administration of HTTLD. Western blot results showed that the protein expression of ZO-1, Occludin, and Claudin-5 in the model group was reduced compared with that in the sham operation group (<italic>P<</italic>0.01). Compared with the model group, the HTTLD groups could up-regulate the expression of ZO-1, Occludin, and Claudin-5 to varying degrees (<italic>P<</italic>0.05, <italic>P<</italic>0.01), especially the high-dose HTTLD group. ELISA showed that the serum <italic>D</italic>-LAC, DAO, and LPS of the model group were elevated compared with those in the sham operation group (<italic>P<</italic>0.01). Compared with the model group, the HTTLD groups showed reduced <italic>D</italic>-LAC and DAO (<italic>P<</italic>0.05, <italic>P<</italic>0.01), and the medium- and high-dose HTTLD groups showed reduced LPS (<italic>P<</italic>0.05, <italic>P<</italic>0.01), especially the high-dose HTTLD group. Conclusion:After cerebral ischemia/reperfusion, the rats showed damaged brain tissues, neurological dysfunction, intestinal mucosal injury, weakened intestinal motility, and destroyed the intestinal mucosal barrier. HTTLD can protect against brain-gut axis injury after cerebral ischemia/reperfusion by reducing the damage on brain tissues and gastrointestinal mucosa, relieving the symptoms of neurological deficits, promoting gastrointestinal motility, improving intestinal barrier function, and reducing the release of intestinal bacterial metabolites or poisons.

14.
Article in Chinese | WPRIM | ID: wpr-906013

ABSTRACT

Objective:To investigate the role of formyl peptide receptor 2 (FPR2) in the inhibitory effects of Buyang Huanwutang (BYHWT) on the oxidative stress and its protective effects on cerebral ischemia-reperfusion in rats. Method:Forty-eight male SD rats were randomly divided into sham group, model group, BYHWT group and BYHWT combined with FPR2 inhibitor (Boc-2) group. In the sham group, only the vessels were isolated. In other groups, the middle cerebral artery occlusion (MCAO) model was constructed using the modified Longa method and reperfused after 2 h of ischemia. BYHWT (16 g·kg<sup>-1</sup>) was given by gavaged twice daily after reperfusion in BYHWT group and BYHWT+Boc-2 group. Boc-2 (0.4 mg·kg<sup>-1</sup>) was injected intraperitoneally 30 min before surgery. Equal volume of saline were given instead in sham and model group. After 24 h of reperfusion, Fluoro-Jade C (FJC) staining was performed to observe the changes in the number of FJC-positive cells. Western blot was performed to detect the expression of apoptosis-related B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X (Bax), and cleaved aspartic acid cysteine proteolytic enzyme-3(Caspase-3). Besides, superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH), and nitric oxide (NO) was measured. The mean fluorescence intensity of nicotinamide adenine dinucleotide phosphate Ⅱ(NADPH) oxidase 2 (NOX2) was examined by immunofluorescence. Result:Compared with sham group, the model group showed increased number of FJC-positive cells (<italic>P</italic><0.01), decreased Bcl-2 expression (<italic>P</italic><0.01), increased Bax and cleaved Caspase-3 expression (<italic>P</italic><0.01), increased NO and MDA content (<italic>P</italic><0.05,<italic>P</italic><0.01), decreased GSH and SOD activities (<italic>P</italic><0.05,<italic>P</italic><0.01), and increased NOX2 expression (<italic>P</italic><0.01). Compared with model group, there were decreased FJC-positive cells (<italic>P</italic><0.01), up-regulated Bcl-2 expression (<italic>P</italic><0.01) with down-regulated cleaved Caspase-3 and Bax (<italic>P</italic><0.05,<italic>P</italic><0.01), decreased NO and MDA (<italic>P</italic><0.05,<italic>P</italic><0.01) with increased GSH and SOD (<italic>P</italic><0.01), and decreased NOX2 expression (<italic>P</italic><0.01) in the BYHWT group. All the above effects were partially blocked by Boc-2. Conclusion:BYHWT can reduce oxidative stress injury and inhibit apoptosis in cerebral ischemia/reperfusion rats, which may be related with the down-regulation of NOX2 expression by FPR2.

15.
Article in Chinese | WPRIM | ID: wpr-905959

ABSTRACT

Objective:To investigate the effect of astragaloside Ⅳ(AST Ⅳ)and Notoginseng total saponins (NTS) combined with bone marrow mesenchymal stem cell (BMSC) transplantation on neural repair and angiogenesis in rats with cerebral ischemia. Method:The rats were randomly divided into a sham operation group, a model group, low- and high-dose AST Ⅳ + NTS groups, a BMSC infusion group, and low- and high-dose BMSC infusion+AST Ⅳ (10 and 20 mg·kg<sup>-1</sup>) + NTS group (25, 50 mg·kg<sup>-1</sup>). BMSCs were isolated and purified by whole bone marrow adherent culture. The positive expression of surface markers of BMSCs (CD29, CD90, CD34, and CD45) was detected by flow cytometry. The focal cerebral ischemia model was established by middle cerebral artery occlusion (MCAO). The PKH26-labeled BMSCs were injected into the tail vein of rats in the BMSC infusion group, once a day. The rats in the combination groups received BMSC injection once a day and intragastric administration of drugs twice a day. Other groups were administered twice a day by gavage. The sham operation group and the model group received the same amount of normal saline. Symptoms and signs of neurological deficits were assessed by the Longa method and the cerebral infarction rate was determined by TTC staining. The survival and vascularization [double positive expression of PKH26/vascular endothelial growth factor (VEGF)] after transplantation of BMSCs were observed by the immunofluorescence method. The protein expression of Ang1 and TGF-<italic>β</italic><sub>1</sub> was measured by Western blot. Result:BMSCs were properly isolated and cultured. The identification of surface markers CD29, CD90, CD34, and CD45 was consistent with the characteristics of BMSCs. The neurological deficit score and cerebral infarction rate of the model group were significantly increased (<italic>P</italic><0.01). All drugs and cell transplantation could alleviate the above pathological changes in varying degrees. The strongest effect was observed in high-dose BMSC infusion+AST Ⅳ+NTS group (<italic>P</italic><0.01), which was superior to those in the AST Ⅳ+NTS groups or the BMSC infusion group. BMSC injection helped cells survive in the ischemic brain tissues and promoted angiogenesis, and this effect could be enhanced by the combination with drugs. After cerebral ischemia, the expression of Ang1 and TGF-<italic>β</italic><sub>1</sub> was increased, and the effect in the BMSC infusion+AST Ⅳ+NTS groups was the strongest (<italic>P</italic><0.01). Conclusion:AST Ⅳ combined with NTS can promote the survival of transplanted BMSCs and facilitate angiogenesis after target repair of damaged blood vessels after cerebral ischemia. The mechanism may be related to the improvement of the local microenvironment in the brain after cerebral ischemia and the promotion of the survival and differentiation of transplanted stem cells.

16.
Article in Chinese | WPRIM | ID: wpr-905924

ABSTRACT

Objective:To observe and compare the protective effects of Tongqiao Huoxue decoction (TQHX) prepared by three methods against cerebral ischemia-reperfusion injury (CIRI), and to explore its mechanism through the glutamate (Glu) metabolic pathway in astrocytes. Method:The male SD rats of SPF grade were subjected to CIRI model induction by the modified middle cerebral artery occlusion method. The model rats were randomly divided into a model group, a sham operation group, and water-decocted, wine-decocted, and alcohol-extracted TQHX (6.3 g·kg<sup>-1</sup>·d<sup>-1</sup>) groups. The rats were treated correspondingly for 7 days. Those in the sham operation group and the model group were treated with an equal volume of normal saline by gavage. After the final treatment, the neurological function of rats was assessed by the modified neurological severity score (mNSS). Hematoxylin-eosin (HE) staining was used to observe the morphological changes of ischemic brain tissues in rats. High-performance liquid chromatography (HPLC) was used to detect glutamate (Glu) in ischemic brain tissues. The expression of glutamate transporter-1 (GLT-1) and glial fibrillary acidic protein (GFAP) and co-expression of glutamine synthetase (GS) and GFAP in ischemic brain tissues were detected by immunofluorescence assay. Western blot was used to detect the protein expression of GFAP, GLT-1, and GS. Result:Compared with the sham operation group, the model group showed increased mNSS (<italic>P</italic><0.01), large necrosis of cerebral cortex in ischemic brain tissues with disordered cell arrangement, obscure boundary, intracellular edema, and inflammatory infiltration, elevated Glu in ischemic brain tissues (<italic>P</italic><0.01), declining GLT-1-GFAP co-expression and GS-GFAP co-expression (<italic>P</italic><0.01), up-regulated expression of GFAP protein, and reduced protein expression of GLT-1 and GS(<italic>P<</italic>0.05,<italic>P<</italic>0.01). Compared with the model group, the TQHX groups showed decreased mNSS (<italic>P<</italic>0.01), relieved injury in the cerebral cortex and hippocampal nerve cells in ischemic brain tissues, reduced Glu expression(<italic>P<</italic>0.05,<italic>P<</italic>0.01), elevated co-expression of GLT-1 and GFAP (<italic>P<</italic>0.05,<italic>P<</italic>0.01), and up-regulated protein expression of GFAP and GLT-1(<italic>P<</italic>0.05,<italic>P<</italic>0.01). The co-expression of GS and GFAP (<italic>P<</italic>0.05,<italic>P<</italic>0.01)and the expression of GS (<italic>P<</italic>0.01)were increased in the wine-decocted and alcohol-extracted TQHX groups. Compared with the water-decocted TQHX group, the alcohol-extracted group showed increased GLT-1-GFAP and GS-GFAP co-expression(<italic>P<</italic>0.05); the wine-decocted and alcohol-extracted TQHX groups exhibited elevated GS protein expression (<italic>P<</italic>0.05); the alcohol-extracted TQHX group displayed declining Glu content (<italic>P</italic><0.01) and increased protein expression of GFAP and GLT-1 (<italic>P<</italic>0.05, <italic>P<</italic>0.01). Compared with the wine-decocted TQHX group, the alcohol-extracted TQHX group showed increased protein expression of GFAP and GLT-1(<italic>P<</italic>0.05,<italic>P<</italic>0.01). Conclusion:TQHX prepared by three methods can improve neurological deficits in CIRI rats. The effect is presumedly achieved by promoting the further activation of astrocytes, increasing the expression of GLT-1 and GS, promoting the clearance of Glu accumulated in the synaptic cleft by astrocytes through the Glu-glutamine (Gln) circulation, and reducing the excitotoxicity of Glu. The alcohol-extracted TQHX group was superior to the water-decocted and wine-decocted TQHX groups in reducing the content of Glu in ischemic brain tissues, promoting the activation of astrocytes, and enhancing the protein expression of GLT-1 and GS.

17.
Article in Chinese | WPRIM | ID: wpr-905918

ABSTRACT

Cerebral ischemia/reperfusion injury (CIRI) is a common feature and the main pathophysiological mechanism of ischemic stroke(IS), which is caused by a blood reperfusion injury in ischemic brain tissues. It can aggravate brain tissue injury and cause irreversible brain damage, seriously affecting the quality of life or even the life of patients. Hence, we must find out the exact mechanism as well as the effective therapeutic drugs and targets for CIRI. The Chinese medicine effective in Xingnao (restoring consciousness) and Kaiqiao (opening orifices) has been widely used in the treatment of CIRI and serves as a classic therapy for IS. In recent years, scholars have conducted extensive and in-depth studies on the mechanism and therapeutic targets of Chinese medicine in Xingnao and Kaiqiao. They found that those drugs could interfere with a series of changes after IS and achieve the remarkable curative effect. This study summarized the effect and mechanism of Chinese medicine in Xingnao and Kaiqiao in the treatment of CIRI, including reducing the inflammatory response and oxidative stress, alleviating brain edema and the toxicity of excitatory amino acids, reducing cell apoptosis, promoting angiogenesis and neurovascular remodeling, and improving blood-brain barrier injury. It is expected to provide references to clarify the mechanism and important targets of those drugs in resisting CIRI and ideas for the in-depth investigation and application of brain protection of Chinese medicine in Xingnao and Kaiqiao.

18.
Article in Chinese | WPRIM | ID: wpr-905307

ABSTRACT

Objective:To observe the interactive effects of early exercise and γ-aminobutyric acid (GABA) receptor antagonists on the neurologic function and brain-derived neurotrophic factor (BDNF) expression in the cerebral ischemia-reperfusion (I/R) rat model. Methods:Male Sprague-Dawley rats were divided into sham (n = 10), I/R (n = 10), exercise (EX) (n = 10), pentetrazol (PTZ) (n = 10) and pentetrazol plus exercise (PTZEX) (n = 10) groups. All the rats, except the sham group, accepted middle cerebral artery occlusion (MCAO) for one hour and reperfused. Since two days after reperfusion, PTZ and PTZEX groups accepted PTZ, a GABA receptor antagonist, 0.25 mg/kg peritoneal injection, once a day for five days; while EX and PTZEX groups ran on a treadmill, 30 minutes a day for five days. Seven days after reperfusion, all the rats were assessed with neurobehavioral score, the infarct volumes were assessed with TTC, and BDNF expression in ischemic penumbra was detected with reverse transcription polymerase chain reaction and ELISA. Results:Compared with I/R group, the neurobehavioral scores of EX group, PTZ group and PTZEX group improved, the volumes of cerebral infarction reduced (P < 0.05), and PTZEX group was the best (P < 0.05). The expression of BNDF was the most in PTZEX group (P < 0.05). Conclusion:Early exerxise combined with PTZ could promote the recovery of neurologic function in I/R rats, which may be related to the up-regulation of BDNF.

19.
Article in Chinese | WPRIM | ID: wpr-905241

ABSTRACT

Objective:To explore the effects of enriched environment on neurological function in cerebral ischemia-reperfusion injury rats and the glucose metabolism in ischemic penumbra. Methods:A total of 72 adult male Sprague-Dawley rats were randomly divided into sham group (n = 24), model group (n = 24) and enriched environment group (n = 24). The latter two groups suffered cerebral ischemia 60 minutes and reperfused with modified Longa's method. The enriched environment group was fed in enriched environment after operation. All the rats were assessed with modified Neurological Severity Score (mNSS) before, and one, seven, 14, 21 and 28 days after operation. One and 28 days after operation, twelve rats from each groups were sacrificed after mNSS assessment, respectively. The histopathology was observed with HE staining. The expressions of hypoxia-inducible factor-1α (HIF-1α), glucose transporter 1 (GLUT1) and 6-phosphofructo-2-kinase/fructose-2-bisphosphatase 3 (PFKFB3) in ischemic penumbra were determined with reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) and Western blotting. The levels of ATP, ADP and AMP in ischemic penumbra were measured with high performance liquid chromatography (HPLC) and the energy charge (EC) was calculated. Results:Compared with the model group, the scores of mNSS decreased in the enriched environment group since 14 days after operation (P < 0.05). The cells in the penumbra presented edema, nuclear pyknosis marginalization, vacuolar arrangement and other pathological changes in the model group and the riched environment group one day after operation; while compared with the model group, the levels of ATP and EC decreased and the mRNAs and protein expression of HIF-1α, GLUT1 and PFKFB3 increased in the enriched environment group (P < 0.05). The pathology improved in the riched environment group compared with that in the model group 28 days after operation; while the mRNAs and protein expression of HIF-1α, GLUT1 and PFKFB3 increased, as well as the levels of ATP and EC (P < 0.05). Conclusion:Enriched environment can promote the recovery of neurological function in rats after cerebral ischemia-reperfusion, which may associate with promoting expression of HIF-1α and downstream GLUT1 and PFKFB3, and improving glucose metabolism.

20.
Article in English | WPRIM | ID: wpr-922098

ABSTRACT

OBJECTIVE@#To investigate the synergistic effect of Naoxintong Capsule (NXTC, ) and Guhong Injection (GHI, ) on cerebral ischemia-reperfusion (I/R) injury.@*METHODS@#Forty-eight Sprague-Dawley rats were divided into 6 groups: control group, oxygen and glucose deprivation (OGD) group, nimodipine group (9.375 mg/kg), NXTC group (0.5 g/kg), GHI group (5 mL/kg) and NXTC+GHI group (0.5 g/kg NXTC+5 mL/kg GHI), after the onset of reperfusion and once per day for the following 7 days. Blood was collected 1 h after final administration, and the sera were collected. Cultured primary rat brain microvascular endothelial cells (rBMECs) were subjected to OGD to establish a cell injury model. Untreated rBMECs were used as blank control. The cell counting kit-8 assay was used to assess cell viability using the sera. Malondialdehyde (MDA) and superoxide dismutase (SOD) levels were assessed using an enzyme-linked immunosorbent assay. Apoptosis was evaluated after Hoechst33342 staining using fluorescence microscopy and flow cytometry. JC-1 staining was performed to assess changes in mitochondrial membrane potential.@*RESULTS@#Statistical analysis indicated that more than 95% of the cells were rBMECs. Compared with the OGD group, the cellular morphology of the all drug delivery groups improved. In particular, the combined drug group had the most significant effect. Compared with the OGD group, all drug intervention groups induced a decrease in the apoptotic rate of rBMECs, increased the SOD levels, and decreased the MDA levels (all P<0.01). Compared with the mono-therapy groups, the NXTC+GHI group exhibited a significant improvement in the number of apoptotic rBMECs (P<0.01). All drug intervention groups showed different degrees of increase in membrane potential, and the NXTC+GHI group was higher than the NXTC or GHI group (P<0.01).@*CONCLUSION@#The combinationa application of NXTC and GHI on cerebral I/R injury clearly resulted in protective benefits.


Subject(s)
Animals , Apoptosis , Brain , Brain Ischemia/drug therapy , Drugs, Chinese Herbal , Endothelial Cells , Glutamine/analogs & derivatives , Plant Extracts , Rats , Rats, Sprague-Dawley , Reperfusion Injury/drug therapy
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