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1.
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.

2.
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.

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 ; 54(10): e11028, 2021. tab, graf
Article in English | LILACS | ID: biblio-1285653

ABSTRACT

Engeletin is a natural derivative of Smilax glabra rhizomilax that exhibits anti-inflammatory activity and suppresses lipid peroxidation. In the present study, we sought to elucidate the mechanistic basis for the neuroprotective and pro-angiogenic activity of engeltin in a human umbilical vein endothelial cells (HUVECs) oxygen-glucose deprivation and reoxygenation (OGD/R) model system and a middle cerebral artery occlusion (MCAO) rat model of cerebral ischemia and reperfusion injury. These analyses revealed that engeletin (10, 20, or 40 mg/kg) was able to reduce the infarct volume, increase cerebral blood flow, improve neurological function, and bolster the expression of vascular endothelial growth factor (VEGF), vasohibin-2 (Vash-2), angiopoietin-1 (Ang-1), phosphorylated human angiopoietin receptor tyrosine kinase 2 (p-Tie2), and platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31) in MCAO rats. Similarly, engeletin (100, 200, or 400 nM) markedly enhanced the migration, tube formation, and VEGF expression of HUVECs in an OGD/R model system, while the VEGF receptor (R) inhibitor axitinib reversed the observed changes in HUVEC tube formation activity and Vash-2, VEGF, and CD31 expression. These data suggested that engeletin exhibited significant neuroprotective effects against cerebral ischemia and reperfusion injury in rats, and improved cerebrovascular angiogenesis by modulating the VEGF/vasohibin and Ang-1/Tie-2 pathways.


Subject(s)
Animals , Rats , Reperfusion Injury/prevention & control , Brain Ischemia/prevention & control , Infarction, Middle Cerebral Artery , Endothelial Cells , Flavonols , Angiopoietin-1 , Vascular Endothelial Growth Factors , Vascular Endothelial Growth Factor A , Glycosides
5.
Article in Chinese | WPRIM | ID: wpr-909603

ABSTRACT

OBJECTIVE To explore the effect of total flavonoids of Rhododendra simsii (TFR) on improving cerebral ischemia/reperfusion injury (CIRI) and its relationship with STIM/Orai-regulated operational Ca2+influx (SOCE) pathway. METHODS Oxygen-glucose deprivation/reoxygenation (OGD/R) PC12 cells were used to simulate CIRI in vitro, and the intracellular Ca2+ concentration and apoptosis rate of PC12 cells were detected by laser confocal microscope and flow cytometry, respectively. The regulation of STIM/Orai on SOCE was analyzed by STIM/Orai gene silencing and STIM/Orai gene overexpression. The CIRI model was established by MCAO in SD rats. The activities of inflammatory cyto?kines IL-1, IL-6 and TNF-αin serum were detected by ELISA. The pathological changes of ischemic brain tissue and the infarction of rat brain tissue were detected by HE staining and TTC staining. The protein and mRNA expression levels of STIM1, STIM2, Orai1, caspase-3 and PKB in brain tissue were detected by Western blotting and RT-qPCR, respectively. RESULTS The results of in vitro experiment showed that the fluorescence intensity of Ca2+ and apoptosis rate in PC12 cells treated with TFR were significantly lower than those in OGD/R group, and this trend was enhanced by SOCE antagonist 2-APB. STIM1/STIM2/Orai1 gene silencing significantly reduced apoptosis and Ca2+overload in OGD/R model, while TFR combined with overexpression of STIM1/STIM2/Orai1 aggravated apoptosis and Ca2+overload. In the in vivo experiment, TFR significantly reduced the brain histopathological damage, infarction of brain tissue, the contents of IL-1, IL-6 and TNF-α in the serum in MCAO rats and down-regulated the expression of STIM1, STIM2, Orai1 and caspase-3 protein and mRNA in the brain tissue, and up-regulated the expression of PKB. The above effects were enhanced by the addition of 2-APB. CONCLUSION The above results indicate that TFR may reduce the contents of inflammatory factors and apoptosis, decrease Ca2+ overload and ameliorate brain injury by inhibiting SOCE pathway mediated by STIM and Orai, suggesting that it has a protective effect against subacute CIRI.

6.
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
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.
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.

9.
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.

10.
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.

11.
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.

12.
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.

13.
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.

14.
China Pharmacy ; (12): 1811-1817, 2021.
Article in Chinese | WPRIM | ID: wpr-886272

ABSTRACT

OBJECTIVE:To obs erve the protective effect of protoca techuic aldehyde(PAL)on neurovascular unit (NVU) homeostasis damage in rats after cerebral ischemia-reperfusion injury (CIRI). METHODS :SD rats were randomly divided into sham operation group ,model group ,PAL high-dose and low-dose groups (10,20 mg/kg),with 11 rats in each group. Administration groups were given relevant medicine intragastrically. Sham operation group and model group were given the same volume of normal saline intragastrically ,10 mL/kg once a day ,for 5 days. After last administration ,CIRI model was induced by suture method ;the ultrastructural changes of NVU were observed by transmission electron microscope. Western blot assay was used to detect the expression of NUV related proteins (MAP-2,GFAP,AQP-4)in cerebral tissue. Immunofluorescence staining was used to observe the positive expression of above proteins in cerebral cortex. RESULTS :Compared with sham operation group , blood-brain barrier (BBB)structure of model group was destroyed severely ,the vascular lumen became narrower ,lateral edema of endothelial cells was severe ,and the thickness of basement membrane varied ;the nuclei of neurons were pyknosis and there was a large area of edema in the surrounding tissues ;the structure of glial cells was seriously damaged ,the cell body was shrunk and organelles were lost ;protein expression (or positive expression )of MAP- 2 in brain tissue (or cerebral cortex )were significantly decreased (P<0.05 or P<0.01),while protein expression (or positive expression ) of GFAP and AQP- 4 were increased significantly(P<0.01). After PAL intervention ,the rats had less BBB damage ,and the morphology of vascular lumen and basement membrane were not completely destroyed ;the damage of neurons was alleviated ,the pyknosis of neurons was decreased , the chromatin was homogeneous and the heterochromatin was decreased;the damage of glial cell structure was alleviated protein expression of GFAP and AQP- 4(except for low-dose group) in cerebral tissue and positive expression of MAP- 2 and GFAP protein in cerebral cortex were reversed @qq.com significantly (P<0.05 or P<0.01). CONCLUSIONS :PAL can protect the stability of NVU from damage in CIRI model rats; the mechanism may be related to up-regulating the expression of MAP- 2 protein in cerebral cortex and down-regulating the expression of GFAP and AQP- 4 protein in brain tissue.

15.
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.

16.
Article in Chinese | WPRIM | ID: wpr-828101

ABSTRACT

OBJECTIVE@#To investigate the protective effects of dexmedetomidine (DEX) against cerebral ischemia/reperfusion (I/R) injury in mice and its relation with mitochondrial fusion and fission.@*METHODS@#Male ICR mice were randomly divided into sham-operated group, I/R group, I/R+DEX group and I/R+DEX+dorsomorphin group. Mouse models of cerebral I/R injury were established by modified thread occlusion of the middle cerebral artery. DEX (50 μg/kg) was injected intraperitoneally at 30 min before cerebral ischemia, which lasted for 1 h followed by reperfusion for 24 h. The neurobehavioral deficits of the mice were evaluated based on Longa's scores. The volume of cerebral infarction was detected by TTC staining. The changes in mitochondrial morphology of the brain cells were observed with transmission electron microscopy. Western blotting was performed to detect the expressions of phosphorylated AMP-activated protein kinase (p-AMPK), mitochondrial fusion protein (Mfn2) and mitochondrial fission protein (p-Drp1) in the brain tissues.@*RESULTS@#DEX pretreatment significantly reduced the neurobehavioral score and the percent volume of cerebral infarction in mice with cerebral I/R injury. Treatment with dorsomorphin (an AMPK inhibitor) in addition to DEX significantly increased the neurobehavioral score and the percent volume of cerebral infarction in the mouse models. Transmission electron microscopy showed that DEX obviously reduced mitochondrial damage caused by cerebral I/R injury and restored mitochondrial morphology of the brain cells, and such effects were abolished by dorsomorphin treatment. Western blotting showed that DEX pretreatment significantly increased the expressions of p-AMPK and Mfn2 protein and decreased the expression of p-Drp1 protein in the brain tissue of the mice, and these changes were also reversed by dorsomorphin treatment.@*CONCLUSIONS@#Preconditioning with DEX produces protective effects against cerebral I/R injury in mice possibly by activating AMPK signaling to regulate mitochondrial fusion and fission in the brain cells.


Subject(s)
Animals , Brain Ischemia , Dexmedetomidine , Male , Mice , Mice, Inbred ICR , Mitochondrial Dynamics , Reperfusion Injury
17.
Acta Anatomica Sinica ; (6): 21-25, 2020.
Article in Chinese | WPRIM | ID: wpr-844545

ABSTRACT

Objective To investigate the effects of the combination of aspirin and nimodipine preconditioning on the prognosis of cerebral ischemia-reperfusion. Methods Eighty healthy male SD rats were randomly divided into sham group, model group, aspirin preconditioning group and aspirin + nimodipine preconditioning group, with 20 rats in each group. The model of cerebral ischemia reperfusion was established. The rats in each group were given intragastric administration for 5 days before the model was established, and the drug was administered daily for 5 days. Sham group and model group were given normal saline; Aspirin preconditioning group was given 50 mg/kg aspirin; Aspirin + nimodipine preconditioning group was given 50 mg/kg aspirin and 10 mg/kg nimodipine. After 2 hours ischemia and 24 hours reperfusion, the animals were neurologically assessed, and then the volume of cerebral infarction was measured by TTC staining. The contents of superoxide dismutase (SOD), malondialdehyde (MDA), thromboxane B2, 6-keto-prostaglandin-la in brain tissue were determined by ELISA. The mRNA expression of Notch 1, Jagged 1 and Hesl in the brain tissues were detected by Real-time PCR, and the expressions of Jaggedl and Hesl, a downstream substance in Notch signaling pathway, were detected by Western blotting. Results The neurological deficit score of the aspirin+nimodipine pretreatment group was significantly lower than model group (P<0. 05) , and the cerebral infarction volume was significantly smaller than other groups. The SOD and 6-keto-prostaglandin 1 in the aspirin pretreatment group and the aspirin plus nimodipine pretreatment group were significantly higher than those in the model group, and the expressions of MDA, thromboxane B2 and thromboxane B2/6-keto-prostaglandin 1 were low. In the model group, the changes in the aspirin + nimodipine pretreatment group were more significant (P<0. 05). The expression levels of Notch 1 , Jagged 1 and Hesl mRNA and protein in the aspirin pretreatment group and aspirin + nimodipine pretreatment group were significantly lower than those in the model group (P<0. 05) , and the expression level of aspirin + nimodipine pretreatment group was lower than that of aspirin pretreatment group (P<0. 05). Conclusion The protective effect of aspirin plus nimodipine is superior to aspirin alone, which can significantly improve cerebral ischemia-reperfusion injury in rats, which may be exerted through influencing notch signaling pathway to achieve brain tissue protection.

18.
Acupuncture Research ; (6): 21-26, 2020.
Article in Chinese | WPRIM | ID: wpr-844211

ABSTRACT

OBJECTIVE: To observe the effect of electroacupuncture (EA) stimulation on the expression of c-Jun terminal kinase(JNK)signaling pathway-related proteins in the hippocampus of vascular dementia (VD) rats, so as to explore its mechanisms underlying improvement of VD. METHODS: Male Sprague-Dawley rats were randomly divided into sham operation, model and EA groups (n=10 rats per group). The VD model was prepared by repeated occlusion of the bilateral common carotid arteries for 10 min and reperfusion for 10 min (3 times in total). The rats in the EA group received EA (2 Hz, 2 mA) at "Dazhui"(GV14),"Baihui"(GV20), and bilateral "Housanli"(ST36) ,"Geshu"(BL17) for 10 min, once daily for 14 days. The learning-memory abi-lity was detected by Morris water maze tests, the distribution of hippocampal neurons detected by Nissl staining, and the apoptosis of hippocampal neurons detected by using TdT-mediated dUTP nick-end labeling (TUNEL) method. The expressions of JNK, phosphorylated JNK (p-JNK), cysteine-containing aspartate-specific proteases-8 (Caspase-8) and Caspase-3 proteins were detected by Western blot. RESULTS: After modeling and compared with the sham operation group, the escape latency was significantly prolonged (P<0.01) and the number of safe-platform quadrant crossing obviously decreased (P<0.01), suggesting a reduction of learning-memory ability. The number of hippocampal neurons was considerably reduced (P<0.01), and that of hippocampal apoptotic neurons remarkably increased in the model group (P<0.01). Whereas, the expression levels of hippocampal apoptosis-related proteins as JNK, p-JNK, Caspase-8 and Caspase-3, as well as the apoptotic index were significantly up-regulated (P<0.01). Following EA intervention, the learning-memory ability was apparently improved (P<0.01), and the number of hippocampal neurons was considerably increased (P<0.01), the hippocampal apoptotic cell number, apoptosis index and the expression levels of JNK, p-JNK, Caspase-8 and Caspase-3 were significantly down-regulated (P<0.01). CONCLUSION: EA intervention can improve the learning-memory ability of VD rats, which may be associated with its effects in reducing hippocampal apoptosis by suppressing JNK signaling pathway.

19.
Article in Chinese | WPRIM | ID: wpr-843894

ABSTRACT

Objective: To observe the effects of butylphthalide on cerebral ischemia-reperfusion injury in rats. Methods: We divided 90 SD rats into sham-operation group, model group, low-dose butylphthalide group, medium-dose butylphthalide group, high-dose butylphthalide group, and ATRA group. Neurological impairment score (NDS) was used to evaluate neurological function. TTC staining was used to calculate the volume of ischemic brain tissue. The xanthine oxidase method was used to detect SOD. The thiobarbituric acid colorimetry was used to detect MDA. ELISA was used to detect IL-6 and TNF-α expressions. The Real-time PCR was used to detect HO-1 gene expression. Western blot was used to detect Nrf2 and HO-1 protein expressions. Results: In low-, medium-, and high-butylphthalide groups, the NDS; volume of ischemic brain tissue; expressions of MDA, IL-6 and TNF-α; 2-△△Ct value of HO-1; protein expressions of Nrf2 and HO-1 were lower than those in model group (P<0.05), but SOD expression was higher than that in model group (P<0.05). The NDS; volume of ischemic brain tissue; expressions of MDA, IL-6 and TNF-α; 2-△△Ct value of HO-1; protein expressions of Nrf2 and HO-1 decreased in a dose-depended manner in low-, medium-, and high-butylphthalide groups, and SOD expression was increased in a dose-depended manner (P<0.05). Conclusion: Butylphthalide can play an antioxidant role by up-regulating Nrf2/HO-1 pathway, which benefits neuroprotective function in cerebral ischemia-reperfusion rats.

20.
Article in Chinese | WPRIM | ID: wpr-872885

ABSTRACT

Objective:To investigate the effect of Xiao Xumingtang combined with super-acupuncture along governor meridian on autophagy-related protein nuclear factor-kappa B p65 (NF-κB p65) in cerebral ischemia-reperfusion model rats, so as to study the relationship between autophagy-related protein NF-κB p65 and brain protection mechanism, and look for the best intervention time point of acupuncture. Method:A total of 152 adult SD male rats were randomly divided into sham operation group, model group, high-dose Xiao Xumingtang group (high-dose drug group), low-dose Xiao Xumingtang group (low-dose drug group) and acupuncture group. There were seven groups including high-dose Xiao Xumingtang + acupuncture group (high acupuncture group) and low-dose Xiao Xumingtang + acupuncture (low acupuncture group). Model group, high-dose drug group, low-dose drug group, and acupuncture group were divided into 4 subgroups according to 30 minutes, 2, 4, 6 h of ischemia-reperfusion, with 6 animals in each group. After successful modeling, according to Zea Longa's neural function score, eligible rats were included into the corresponding groups. The sham operation group only received carotid artery dissection, the model group was only modeled without any treatment, high and low-dose Xiao Xumingtang groups were calculated based on the body surface area of the animal and given 60 g·kg-1·d-1 and 15 g·kg-1·d-1 drug by gavage for treatment, acupuncture was performed to smooth governor meridian and regulate the mind. After 14 days of consecutive treatment, neurological function was scored. Western blot was used to detect the expression of autophagy-related protein NF-κB p65 in rat brain tissue. Result:Compared with the sham operation group, the neurological impairment scores of the model group, the high-dose drug group, the low-dose drug group, the acupuncture group, the high acupuncture group, and the low acupuncture group were significantly increased at each time point (P<0.01). The neurological impairment scores were significantly lower at each time point than those of the high-dose drug group, low-dose drug group, acupuncture group, high acupuncture group, and low acupuncture group (P<0.01), compared with the sham operation group, NF-κB p65 in model group, high-dose drug group, low-dose group, acupuncture group, high acupuncture group and low acupuncture group was significantly increased in the brain tissue at each time point (P<0.01), compared with the model group, the expression of NF-κB p65 protein in the brain tissue of model group, high-dose drug group, low-dose group, acupuncture group, high acupuncture group and low acupuncture group was decreased at each time point (P<0.05), particular in the high acupuncture group (P<0.01). Conclusion:Xiao Xumingtang combined with ultra-early acupuncture along governor meridian can significantly alleviate neurological impairment in rats with cerebral ischemia-reperfusion model. Xiao Xumingtang combined with ultra-early acupuncture along governor meridian can inhibit cerebral ischemia-reperfusion model rats. The activity of autophagy-related protein NF-κB P65 protects the brain function. There is no significant difference in the brain protective effect of Xiao Xumingtang combined with ultra-early acupuncture along governor meridian within 6 hours.

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