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Objective To elucidate the mechanism of dl-3-N-butylphthalide (NBP) on renal ischemia-reperfusion injury (IRI) in rat models. Methods Forty SD rats were randomly divided into the sham operation group (Sham group), model group (IRI group), NF-κB inhibitor pyrrolidine dithiocarbamate group (PDTC group), low-dose NBP group (NBP-L group) and high-dose NBP group (NBP-H group), with 8 rats in each group. Serum creatinine (Scr), serum cystatin C(Cys-C), blood urea nitrogen (BUN) and serum interleukin (IL)-1β and IL-18 levels were detected in all groups. Pathological injury of renal tissues in each group was observed by Hematoxylin-eosin (HE) staining. The expression levels of inflammatory factors and nuclear factor (NF)-κB signaling pathway and cell pyroptosis-related proteins in renal tissues were measured by Western blot and immunohistochemical staining. Results Compared with the Sham group, renal tissue injury was more severe, and the levels of Scr, Cys-C, BUN and serum IL-1β and IL-18 were all up-regulated in the IRI group. Western blot showed that the relative expression levels of NOD-like receptor protein (NLRP3), Gasdermin D(GSDMD), cysteinyl aspartate specific proteinase (Caspase)-1, IL-18, IL-1β, NF-κB p65 and p-NF-κB p65 proteins were all up-regulated, and immunohistochemical staining revealed that the expression levels of NF-κB p65 and p-NF-κB p65, IL-1β, IL-18 and NLRP3 proteins were all up-regulated in the IRI group. Compared with the IRI group, renal tissue injury was alleviated, and the levels of Scr, Cys-C, BUN and serum IL-18 and IL-1β were down-regulated in the PDTC, NBP-L and NBP-H groups. Western blot showed that the expression levels of NLRP3, GSDMD, Caspase-1, IL-1β, IL-18, NF-κB p65 and p-NF-κB p65 proteins were down-regulated, and immunohistochemical staining indicated that the expression levels of NF-κB p65, p-NF-κB p65, IL-1β, IL-18 and NLRP3 proteins were down-regulated in the PDTC, NBP-L and NBP-H groups, respectively. Compared with the NBP-L group, renal tissue injury was mitigated, and the levels of Scr, Cys-C, BUN, serum IL-18 and IL-1β were all down-regulated in the NBP-H group. Western blot showed the expression levels of NLRP3, GSDMD, Caspase-1, IL-1β, IL-18, NF-κB p65 and p-NF-κB p65 proteins were down-regulated in the NBP-H group. Immunohistochemical staining indicated that the expression levels of NF-κB p65, p-NF-κB p65, IL-1β, IL-18 and NLRP3 proteins were down-regulated in the NBP-H group. Conclusions NBP may down-regulate the activity of NF-κB/NLRP3 signaling pathway and reduce the expression levels of cell pyroptosis-related proteins and inflammatory factors after renal IRI, thereby suppressing cell pyroptosis and alleviating renal IRI.
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OBJECTIVE To study the improvement effect and possible mechanism of N-butylphthalide on inflammatory injury of bone marrow mesenchymal stem cells （BMSCs） in rats. METHODS BMSCs of rats were divided into control group， model group， N-butylphthalide low-concentration， medium-concentration and high-concentration groups （10， 20， 50 μmol/L）. BMSCs were cultured in vitro and lipopolysaccharide （the final concentration of 10 mg/L） was used to establish the inflammatory injury model. After the intervention of N-butylphthalide， the survival rate， apoptotic rate， the contents of tumor necrosis factor α （TNF- α）， interleukin 1β （IL-1β） and IL-6 in cell culture medium， the mRNA expression of nuclear factor-κB（NF-κB） p65， and the protein expressions of caspase-3， B-cell lymphoma 2 （Bcl-2）， Bcl-2 related X protein （Bax） and NF-κB p65 in cells were detected. RESULTS Compared with control group， the survival rate and protein expression of Bcl-2 were decreased significantly in model group （P＜0.05）； the apoptotic rate， contents of TNF-α， IL-1β and IL-6， the mRNA expression of NF-κB p65， and the protein expressions of caspase-3， Bax and NF-κB p65 were increased significantly （P＜0.05）. Compared with model group， above indexes were significantly reversed in all concentration groups of N-butylphthalide （P＜0.05）， in concentration-dependent manner. CONCLUSIONS N-butylphthalide can ameliorate the inflammatory injury of BMSCs induced by lipopolysaccharide， and its mechanism may be related to the inhibition of NF-κB signaling pathway.
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Butylphthalide and ferulic acid exhibit excellent therapeutic effects in ischemic stroke. In this research, twelve 3-n-butylphthalide derivatives were designed by molecular hybridization strategy. The target compounds were obtained by nucleophilic substitution, reduction reaction, esterification reaction and elimination reaction, and the structure was confirmed by 1H NMR, 13C NMR and ESI-MS. All compounds were evaluated for neuroprotective activity against OGD/R-induced neurotoxicity in rat cortical neurons by MTT assay. The compounds with the best neuroprotective activity were biologically evaluated for their ability to inhibit platelet aggregation induced by arachidonic acid (AA) and adenosine diphosphate (ADP) via the Bron method．The results indicate that 7b exhibited potent neurocyte protective activity as well as prominent anti-platelet aggregation activity. Compound 7b has potential to be developed as a drug for ischemic stroke.
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Objective: To study the content variation and chemical composition of Siwu Decoction between mixed decoction and single decoction comprehensively, and then explore variation rule of Siwu Decoction by different decocting methods based on material basis. Methods: Components of Siwu Decoction were identified by LC-MS/MS and an UPLC wavelength switching method for simultaneously determining the contents of multiple compounds in Siwu Decoction was established based on the idea of TCM chemistry holography. The mixed and single decoction samples were prepared and tested. Experimental data was compared to analyze material basis differences and variation rule of Siwu Decoction by different decocting methods. Results: A total of 72 compounds were identified and assigned, 18 compounds were quantitative detected and all of 18 analytes showed good linearity (R2 ≥ 0.999) within the test range. The relative standard deviations of the precision, repeatability and stability were not exceeding 2.0%, and the recoveries were in the range of 97%-105%. Analysis of Siwu Decoction samples showed dissolution of ligustilide, 3-n- butylphthalide, catechin, gallic acid and paeoniflorin was affected by the change of solvent volume and dissolution of aucubin, catechin, oxypaeoniflorin, paeoniflorin and acteoside were higher in mixed decoction than single decoction obviously. Compared to single decoction, the kinds of compounds in mixed decoction did not change significantly but the content showed notable variety. Conclusion: Through the study of chemistry holography, the composition and content of compounds in TCM mixed decoction and herbs single decoction can be compared and analyzed comprehensively to provide a new perspective for the study on the rule of TCM decoction and dissolution. TCM chemistry holography study may become a useful exploration of the TCM quality study.
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Background@#The increased permeability of the blood-brain barrier (BBB) induced by ischemia/hypoxia is generally correlated with alteration of tight junctions (TJs). DL-3-n-butylphthalide (NBP) has been shown to exert neuroprotective effects after ischemic injury. However, few studies have assessed the correlation between NBP and TJs. This study aimed to investigate the potential effect of NBP on the TJ proteins claudin-5, zonula occludens-1 (ZO-1), and occludin during brain ischemia.@*Methods@#A chronic cerebral hypoperfusion (CCH) Sprague-Dawley rat model was established, and NBP (20, 40, or 80 mg/kg, gavage, once a day) treatment was performed for 14 days. NBP (0.1 or 1.0 μmol/L) pre-treatment was applied to an in vitro hypoxia microvascular endothelial cell model (1% O2, 24 h). BBB permeability was assessed by performing the Evans blue assay. The expressions and localization of claudin-5, ZO-1, occludin, phosphorylated/total protein kinase B (p-Akt/Akt), phosphorylated/total glycogen synthase kinase 3β (GSK-3β)/GSK-3β, and β-catenin/β-actin were evaluated by Western blotting or immunofluorescence. Reactive oxygen species (ROS) generation was measured by flow cytometry analysis. TJ ultrastructure was observed by transmission electron microscopy.@*Results@#In CCH rats, treatment with 40 and 80 mg/kg NBP decreased the Evans blue content in brain tissue (9.0 ± 0.9 μg/g vs. 12.3 ± 1.9 μg/g, P = 0.005; 6.7 ± 0.6 μg/g vs. 12.3 ± 1.9 μg/g, P < 0.01), increased the expression of claudin-5 (0.79 ± 0.08 vs. 0.41 ± 0.06, P < 0.01; 0.97 ± 0.07 vs. 0.41 ± 0.06, P < 0.01), and elevated the ZO-1 protein level (P < 0.05) in brain microvascular segments in a dose-dependent manner in comparison with the corresponding values in the model group. There was no significant difference in occludin expression (P > 0.05). In the hypoxia cell model, NBP pre-treatment improved TJ ultrastructure, decreased intracellular ROS level, and increased the expression of claudin-5 (P < 0.01) and ZO-1 (P < 0.01) in comparison with the corresponding values in the hypoxia group. NBP treatment also elevated the relative expression levels of p-Akt/Akt, p-GSK-3β/GSK-3β, and β-catenin/β-actin in comparison with the corresponding values in the hypoxia group (all P < 0.05).@*Conclusion@#NBP improves the barrier function of BBB against ischemic injury by upregulating the expression of TJ proteins, possibly by reducing oxidative stress and activating the Akt/GSK-3β/β-catenin signaling pathway.
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Objective@#The 3-N-butylphthalide (NBP) comprises one of the chemical constituents of celery oil. It has a series of pharmacologic mechanisms including reconstructing microcirculation, protecting mitochondrial function, inhibiting oxidative stress, inhibiting neuronal apoptosis, etc. Based on the complex multi-targets of pharmacologic mechanisms of NBP, the clinical application of NBP is increasing and more clinical researches and animal experiments are also focused on NBP. The aim of this review was to comprehensively and systematically summarize the application of NBP on neurologic diseases and briefly summarize its application to non-neurologic diseases. Moreover, recent progress in experimental models of NBP on animals was summarized.@*Data sources@#Literature was collected from PubMed and Wangfang database until November 2018, using the search terms including "3-N-butylphthalide," "microcirculation," "mitochondria," "ischemic stroke," "Alzheimer disease," "vascular dementia," "Parkinson disease," "brain edema," "CO poisoning," "traumatic central nervous system injury," "autoimmune disease," "amyotrophic lateral sclerosis," "seizures," "diabetes," "diabetic cataract," and "atherosclerosis."@*Study selection@#Literature was mainly derived from English articles or articles that could be obtained with English abstracts and partly derived from Chinese articles. Article type was not limited. References were also identified from the bibliographies of identified articles and the authors’ files.@*Results@#NBP has become an important adjunct for ischemic stroke. In vascular dementia, the clinical application of NBP to treat severe cognitive dysfunction syndrome caused by the hypoperfusion of brain tissue during cerebrovascular disease is also increasing. Evidence also suggests that NBP has a therapeutic effect for neurodegenerative diseases. Many animal experiments have found that it can also improve symptoms in other neurologic diseases such as epilepsy, cerebral edema, and decreased cognitive function caused by severe acute carbon monoxide poisoning. Moreover, NBP has therapeutic effects for diabetes, diabetes-induced cataracts, and non-neurologic diseases such as atherosclerosis. Mechanistically, NBP mainly improves microcirculation and protects mitochondria. Its broad pharmacologic effects also include inhibiting oxidative stress, nerve cell apoptosis, inflammatory responses, and anti-platelet and anti-thrombotic effects.@*Conclusions@#The varied pharmacologic mechanisms of NBP involve many complex molecular mechanisms; however, there many unknown pharmacologic effects await further study.
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6-Bromo-3-n-butylphthalide was obtained by nitration, reduction and diazotization from carboxybenzaldehyde. Twenty hybrids from substituted styrene and 6-bromo-3-n-butylphthalide were synthesized and the structure was confirmed by 1H NMR, 13C NMR and ESI-MS. All compounds were evaluated for neuroprotective activity against OGD/R-induced neurotoxicity in rat cortical neurons by MTT assay. The mechanism of neuroprotection was investigated by Western blot analyses. The results indicated that most of these compounds had a potent neuroprotective activity (All animal experiments were approved by the Experimental Animal Ethics Committee of Anhui University of Chinese Medicine), especially 10h and 10i showed significant effects, which may play a neuroprotective role by activating the PI3K/Akt signaling pathway.
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Objective@#To explore the effects of n-butylphthalide (NBP) on mitochondria in hippocampus and learning and memory abilities in rats with chronic alcoholism.@*Methods@#60 male SD rats were randomly divided into three groups on average, including normal group, model group and treatment group, with 20 rats in each group.Rats of model group and treatment group are given 6% (V/V) alcohol solution continuously for 28 d to establish the model of chronic alcoholism.Rats in the treatment group were given butylphthalide for 14 days from the fourteenth day after giving alcohol solution.The Y type electric maze was used to test the learning and memory ability of rats, the content of H2S in the hippocampus and the activity of mitochondrial ATP enzyme were measured by spectrophotometry, and the protein expression of F-actin was detected by Western blot.@*Results@#Compared with the normal group, the learning and memory ability of the rats in the model group were decreased, the content of H2S in the hippocampus were increased, and the activity of mitochondrial ATP enzyme and the expression of F-actin protein were decreased, and most of the mitochondria were damaged under the electron microscope.The training times of the rats in treatment group(61.88±3.61)was lower than that of the model group(82.19±4.87), the ability of learning and memory was improved(P<0.05). Compared with the model group ((1.50±0.07)U/mgprot, (0.08±0.01)), the activity of the mitochondrial ATP enzyme((1.84±0.11)U/mgprot) and the level of F-actin protein(0.12±0.01)in rat hippocampus of treatment group were increased, the difference was statistically significant(both P<0.05). The level of H2S in rat hippocampus of the treatment group ((34.56±2.47) nmol/g) was lower than that of the model group ((44.55±3.71) nmol/g), the difference was statistically significant(P<0.05). Compared with model group, the mitochondrial damage of the hippocampus in the treatment group was improved under electron microscope.@*Conclusion@#NBP can abate mitochondrial damage and improve learning and memory abilities in chronic alcoholism rats.
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Objective To explore the effects of N-butylphthalide on the expressions of ZO-1 and claudin-5 in blood-brain barrier (BBB) in rats with acute carbon monoxide (CO) poisoning. Methods A total of 144 adult healthy male Sprague-Dawley (SD) rats were randomly divided into normal control group, CO poisoning group, and NBP treatment group, with 48 rats in each group. The acute CO poisoning model was reproduced in hyperbaric oxygen chamber, and all model rats were given hyperbaric oxygen therapy once daily. The rats in the normal control group were free to breathe fresh air. The rats in NBP treatment group were administered orally NBP 60 mg/kg twice a day at 2 hours after poisoning until death. The rats in normal control group and CO poisoning group were treated with equal amount of pure olive oil. Four rats were sacrificed from each group at 1, 3, 7, 14 days after model reproducing, respectively. The changes in ultrastructure of BBB were observed under transmission electron microscope. The expressions of ZO-1 and claudin-5 proteins were determined by immunofluorescence staining and Western Blot. The localization of the two target proteins was observed by immunofluorescence double staining. The correlation between the two proteins was analyzed by linear regression. Results The ultrastructure of BBB was normal in normal control group, some ZO-1 and a large number of claudin-5 positive cells were observed. The ultrastructure of BBB was seriously injured, ZO-1 and claudin-5 positive cells in brain tissue were significantly decreased, and the expressions of ZO-1 and claudin-5 proteins in brain tissue at 1 day after poisoning in CO poisoning group were significantly lower than those of normal control group (ZO-1 protein:3.38±0.30 vs. 24.50±5.62, claudin-5 protein: 11.38±0.93 vs. 46.35±6.88, both P < 0.05), and although gradually restored, they were maintained at relatively lower levels until 14 days as compared with those in normal control group (ZO-1 protein: 10.35±0.80 vs. 24.63±3.57, claudin-5 protein: 32.35±3.11 vs. 46.43±7.20, both P < 0.05). NBP treatment could significantly alleviate the ultrastructure injury of BBB induced by acute CO poisoning, the amount of ZO-1 and claudin-5 positive cells in brain tissue were significantly increased, as well as the expressions of ZO-1 and claudin-5 proteins were significantly increased, which were significantly higher than those of CO poisoning group from 1 day and 3 days on, respectively (1-day ZO-1 protein: 7.57±0.69 vs. 3.38±0.30, 3-day claudin-5 protein:20.46±1.42 vs. 11.43±0.86, both P < 0.05), and which showed an increase tendency with time prolongation. The results of immunofluorescence double staining showed that ZO-1 and claudin-5 proteins could not only coexist in the same cell, but also could be expressed separately in different cells. Linear regression analysis showed the positive correlation between the expressions of ZO-1 and claudin-5 proteins in brain tissue of rats with acute CO poisoning (R2= 0.917, P = 0.022). Conclusion NBP could markedly improve the ultrastructure and functional integrity of BBB through up-regulating the expressions of ZO-1 and claudin-5 proteins, and then reduce brain damage caused by CO poisoning.
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Objective To investigate the effect of Dl-3-n-butylphthalide on acute cerebral infarction.Methods In Department of Neurology in the Fifth Hospital of Wuhan from March 2013 to June 2014,100 cases of patients with first onset of acute cerebral infarction were recruited.The participants were divided into 2 groups (control group and treatment group) randomly,with 50 participants in each group.Besides general treatment,the patients of treatment group received intravenous injection of Dl-3-n-butylphthalide in acute phase and orally took soft capsule of Dl-3-n-butylphthalide in recovery phase.All the patients were followed up for 24 weeks.Neurological function and general cognition were assessed separately by national institute of health stroke scale (NIHSS),and mini mental state examination (MMSE) was applied to assess overall cognitive function.Results NHISS score was gradually decreased and MMSE score was increased in both groups.As compared with the control group,NIHSS score and MMSE score were changed significantly in the treatment group.From first onset to 24 weeks after treatment,NHISS score was decreased by 30％ in the control group and 44％ in the treatment group;MMSE score was increased by 17％ in the control group and 32％ in the treatment group.Conclusion Sequential therapy with Dl-3-n-butylphthalide improves neurological function and general cognition faster and more significant for patients with acute cerebral infarction.
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Objective Isolation , purification and structure identification of phthalides from A pium graveolens L . Methods Silica gel column and preparative TLC chromatography were used to isolate and purify the phthalide compounds .The structures of those compounds were determined through spectral analysis .Results Three pure compounds were identified as 3-n-butylphthalide ,sedanenolide and sedanolide .Conclusion Three pure phthalide compounds can be used as analytical reference standard for quantitative analysis of A .graveolens .
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Objective:To probe the therapeutic effects of Butylphthalide Injection in the elderly patients with acute cerebral infarction (ACI) and its influence in cerebral hemodynamics and cerebral vascular reserve (CVR),and to clarify the pharmacological action mechanism of butylphthalide in treatment of ACI.Methods:A total of 100 cases of elderly patients with ACI were selected as the subjects and divided into observation group and control group according to the serial number on admission.Fifty cases were included in each group.The patients in control group were treated with the conventional treatment, while the patients in observation group were treated with Butylphthalide Injection on the basis of the conventional treatment.The National Institute of Health Stroke Scale (NIHSS) score, the brain hemodynamics indexes of the peak velocity (Vp), the mean velocity (Vm) and the differences of the velocity (DVp, DVm) as well as pulsatility index (PI), CVR of bilateral middle cerebral artery (MCA)of the patients in two groups were observed and compared.The therapeutic effects of the patients in two groups were evaluated and compared.Results:The NIHSS score of the patients in observation group after treatment was significantly lower than that in control group (t=15.420, P<0.05).The therapeutic effects and the clinical efficiency of the patients in observation group were significantly better than those in control group (U=2.225, χ2=5.005, P<0.05).The Vp and Vm of the patients in observation group after treatment were significantly higher than those in control group(t=10.819,t=7.259, P<0.05)and the DVp and DVm were significantly lower than those in control group (t=16.438,t=19.055, P<0.05).The CVR of the patients in observation group after treatment was significantly higher than that in control group(t=6.884, P<0.05)and the PI was significantly lower than that in control group (t=4.979, P<0.05).Conclusion:Butylphthalide Injection can effectively correct the abnormality of brain hemodynamics in the ACI patients, enhance the ability of body in maintaining the stability of cerebral vascular perfusion, improve the neurological symptoms in the patients with ACI, and improve the therapeutic effects.
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Objective To investigate the effects of DL-3-n-Butylphthalide(NBP)on proliferation and apoptosis of 1-methyl-4-phenyl-pyridinium (MPP +)-induced SH-SY5Y cells, and mechanisms via mixed lineage kinase 3 (MLK3) signaling pathway. Methods The SH-SY5Y cells were divided into control group,MPP+group,NBP group and URMC-099 group,that cultured normally,with 1 mmol/L MPP+for 24 hours,with 10μmol/L NBP for 3 hours and then with MPP+for 24 hours,and with 200 nmol/L MLK3 inhibitor URMC-099 for 3 hours and then with MPP+for 24 hours,respectively.The morphology of SH-SY5Y cells was observed under inverted phase contrast mi-croscope and the survival rate was measured with 3-(4,5-Cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assays.The apoptosis was quantified under flow cytometry with Annexin V/PI fluorescence staining,and the nuclear morphology was observed with Hoechst 33342 staining.The expression of phosphorylated protein of MLK3(p-MLK3),c-Jun N-terminal kinase(p-JNK),extra cellular regulated protein ki-nases(p-ERK1/2)were detected with Western blotting.Results Compared with the control group,the survival rate reduced and apoptosis in-creased in MPP+group(P<0.05),with the increase of p-MLK3 and p-JNK and decrease of p-ERK1/2 d(P<0.05).Compared with MPP+group,the survival rate increased and apoptosis reduced in both NBP and URMC-099 groups(P<0.05),with the decrease of p-MLK3 and p-JNK and increase of p-ERK1/2(P<0.05).Conclusion NBP can decrease the apoptosis and promote the proliferation of SH-SY5Y cells in-duced by MPP+,which may be associated with inhibiting MLK3 signaling pathway,and regulating the downstream p-JNK and p-ERK1/2.
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Objective To investigate the effects of N-butylphthalide (NBP) on cognitive function in acute severe carbon monoxide (CO) poisoning rats and its mechanism. Methods 120 health Sprague-Dawley (SD) rats were randomly divided into three groups (n = 40): normal control group (NC group), CO poisoning group (CO group) and NBP treatment group (NBP group). The acute severe CO poisoning model was established in a hyperbaric oxygen chamber by intoxicated with 1 000 ×10-6CO for 40 minutes, followed with 3 000 ×10-6CO for another 20 minutes, and then received hyperbaric oxygen therapy 1.5 hours once a day until sacrificed. Rats in NBP group were administered orally NBP 60 mg/kg for 2 times daily until death. NC group and CO group were treated with equal amount of pure olive oil. Four rats in each group were taken from 1, 3, 7, 14, 30 days after model setup, respectively. The cognitive function score was assessed by Morris water maze test. The changes in ultrastructure of hippocampus were observed under transmission electron microscope. The expressions of calpain 1 and Ca2+/calmodulin dependent protein kinase Ⅱ(CaMK Ⅱ) in hippocampus of brain tissue were detected by immunofluorescence staining, and the localization of the two target proteins in neurons was observed by immunofluorescence double staining. Results Compared with NC group, the escape latency at 1 day after poisoning in CO group was significantly prolonged (s: 55.6±3.2 vs. 44.5±3.5, P < 0.05), and the times of the platform crossing was significantly decreased (times: 1.3±0.8 vs. 6.6±1.2, P < 0.05);the ultrastructure of hippocampus was obviously injured; the protein expressions of calpain 1 and CaMK Ⅱ in brain tissue were significantly increased at 1 day after CO poisoning [calpain 1 (A value): 41.24±5.21 vs. 6.44±1.13, CaMK Ⅱ (A value): 56.19±5.04 vs. 9.84±1.53, both P < 0.05], and the protein expression of calpain 1 reached the peak at 3 days (A value: 59.34±6.11), the protein expression of CaMK Ⅱ reached the peak at 1 day (A value:56.19±5.04). Compared with CO group, the cognitive function was significantly improved in NBP group in the late stage of poisoning [7-30 days, escape latency (s): 40.3±1.9 vs. 49.1±3.1 at 7 days, 30.1±2.9 vs. 39.4±3.1 at 30 days;times of the platform crossing (times): 2.8±1.0 vs. 1.0±0.9 at 14 days, 3.2±0.8 vs. 1.0±0.9 at 30 days, all P < 0.05];the degree of injury of hippocampal neuron was relatively slight; the protein expression of calpain 1 in brain tissue was significantly decreased from 3 days after CO poisoning (A value: 39.63±3.03 vs. 59.34±6.11, P < 0.05), and the protein expression of CaMK Ⅱ was significantly decreased from 1 day after CO poisoning (A value: 42.22±3.84 vs. 56.19±5.04, P < 0.05). Immunofluorescence double staining suggested that calpain 1 and CaMK Ⅱ protein could not only coexist in the same cell, but also could be expressed separately in different cells. Linear regression analysis showed that the expression of calpain 1 and CaMK Ⅱ was positively correlated (R 2= 0.852, P = 0.002). Conclusions NBP treatment could maintain ultrastructure integrity of hippocampus, balance the expression levels of calpain 1 and CaMK Ⅱproteins, and significantly improve cognitive impairment induced by CO poisoning, thus play a protective role against hippocampus damage in rats with acute severe CO poisoning.
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Objective:To study the effect of 3-n-butylphthalide (NBP) on the expression of brain-derived neurotrophic factor (BDNF) in the hippocampal CA1 region of the vascular dementia (VD) rats,and to explore its protective effect on VD.Methods:Eighty healthy Wistar rats were equally and randomly assigned to sham operation group,NBP control group (sham operation + NBP injection),VD group (VD models),NBP treatment group (VD models + NBP injection) (n=20).Each group was divided into four subgroups (n =5):1,2,4,and 8 weeks after operation groups.The VD rat models were established by using permanent bilateral common carotid artery ligation.After consciousness,the rats in NBP treatment group and NBP control group were intraperitoneally injected with 5 mg · kg-1 · d-1 NBP for consecutive 7 d.The rats in VD and sham operation groups were intraperitoneally injected with 0.2 mL · d-1 saline for consecutive 7 d.At 1,2,4,and 8 weeks after operation,the rats in each group were decapitated.The brains were obtained,and then the hippoeampus tissues were isolated.The BDNF expression levels in the hippocampal CA1 region were determined using real-time quantitative PCR and immunohistochemistry methods.Results:At 2,4,and 8 weeks after s operation,the expression levels of BDNF mRNA in the hippocampus of the rats in VD group were significantly higher those in sham operation group (P＜ 0.05);at 4 and 8 weeks after operation,the expression levels of BDNF mRNA in the hippocampus of the rats in NBP treatment groups were significantly higher than that in VD group (P＜ 0.05).The immunohistochemistry results showed that the expression level of BDNF protein in the hippocampal CA1 region of the rats in VD group was higher than that in sham operation group at 4 weeks after operation (P＜ 0.05);the expression level of BDNF protein in the hippocampal CA1 region of the rats in NBP treatment group was higher than that in VD group at 8 weeks after operation (P＜0.05).Conclusion:The BDNF expression is increased in the hippocampal CA1 region of the rats with VD after the neurons were injured by ischemia.NBP can increase the BDNF expression level in the hippocampal CA1 region of the VD rats and protect the nerves.
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Objective:To study the effect of 3-n-butylphthalide (NBP) on the expression of brain-derived neurotrophic factor (BDNF) in the hippocampal CA1 region of the vascular dementia (VD) rats,and to explore its protective effect on VD.Methods:Eighty healthy Wistar rats were equally and randomly assigned to sham operation group,NBP control group (sham operation + NBP injection),VD group (VD models),NBP treatment group (VD models + NBP injection) (n=20).Each group was divided into four subgroups (n =5):1,2,4,and 8 weeks after operation groups.The VD rat models were established by using permanent bilateral common carotid artery ligation.After consciousness,the rats in NBP treatment group and NBP control group were intraperitoneally injected with 5 mg · kg-1 · d-1 NBP for consecutive 7 d.The rats in VD and sham operation groups were intraperitoneally injected with 0.2 mL · d-1 saline for consecutive 7 d.At 1,2,4,and 8 weeks after operation,the rats in each group were decapitated.The brains were obtained,and then the hippoeampus tissues were isolated.The BDNF expression levels in the hippocampal CA1 region were determined using real-time quantitative PCR and immunohistochemistry methods.Results:At 2,4,and 8 weeks after s operation,the expression levels of BDNF mRNA in the hippocampus of the rats in VD group were significantly higher those in sham operation group (P＜ 0.05);at 4 and 8 weeks after operation,the expression levels of BDNF mRNA in the hippocampus of the rats in NBP treatment groups were significantly higher than that in VD group (P＜ 0.05).The immunohistochemistry results showed that the expression level of BDNF protein in the hippocampal CA1 region of the rats in VD group was higher than that in sham operation group at 4 weeks after operation (P＜ 0.05);the expression level of BDNF protein in the hippocampal CA1 region of the rats in NBP treatment group was higher than that in VD group at 8 weeks after operation (P＜0.05).Conclusion:The BDNF expression is increased in the hippocampal CA1 region of the rats with VD after the neurons were injured by ischemia.NBP can increase the BDNF expression level in the hippocampal CA1 region of the VD rats and protect the nerves.
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Objective To investigate the clinical efficacy of dl-3n-butylphthalide and its effect on NT-proBNP levels and inflammatory cytokines of patients with acute cerebral infarction.Methods Total 118 patients with acute cerebral infarction in Hengshui Fifth People's Hospital were randomly divided into control group and observation group.Sixty patients in control group were treated with conventional medication,and 58 patients in observation group were treated with dl-3n-butylphthalide injection based on conventional medication.The NT-proBNP levels,score of neurological deficits (NIHSS),inflammatory cytokines including hypersensitive C-reactive protein (hs-CRP),interleukin-6 (IL-6),and tumor necrosis factor-cα (TNF-α) before and after 7 and 14 d of treatment,and adverse reaction were observed and analyzed.Results Before treatment,differences in the NT-proBNP levels,NIHSS score,hs-CRP,IL-6,and TNF-α between two groups were not obvious.After 7 and 14 d of treatment,NT-proBNP levels in observation group were significantly lower than those in control group (P ＜ 0.05).And after treatment for 14 d,NIHSS score in observation group were conspicuously lower than those in control group (P ＜ 0.05).Additionally,the levels of hs-CRP,IL-6,and TNF-α in observation group were markedly lower than those in control group (P ＜ 0.05).Conclusion D1-3n-butylphthalide injection has a good ability in inhibiting NT-proBNP levels and inflammatory cytokines,and improves the neurological function of patients with acute cerebral infarction.
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@#A series of hydrogen sulfide-releasing derivatives of open ring 3-n-butylphthalide(5a-5f)were designed, synthesized, and their structures were confirmed by MS and 1H NMR. The inhibitory activity of the target compounds against adenosine diphosphate(ADP)and arachidonic acid(AA)-induced platelet aggregation was evaluated in vitro by Born′s turbidimetric assay. In comparison with 3-n-butylphthalide(NBP), compound 5e possessed better antiplatelet aggregation activity. Therefore, it may be utilized as a lead compound for further investigation.
ABSTRACT
Objective To compare the oil yield of celery seeds and the contents of 3-n-butylphthalide and the total phthalocyanine lactones of celery seed oil extracted by different methods. Methods Three routine extraction methods involving organic solvent extraction, Soxhlet extraction, steam distillation extraction, as well as subcritical extraction method and supercritical fluid extraction method were used to extract the celery seed oil. The contents of 3-n-butylphthalide and total phthalocyanine lactones were respectively detected by high performance liquid chromatography(HPLC) and ultraviolet visible spectrophotometry. Results The ranges of oil yield and the contents of 3-n-butylphthalide and total phthalocyanine lactones of celery seed oil extracted by different methods were 0.30%-20.02%, 1.40%-10.13%, 4.74%-17.65%, respectively, indicating obvious differences. Conclusion With R134a and butane as the solvents, the subcritical extraction method is better than other extraction methods for the extraction of 3-n-butylphthalide. With dimethyl ether as the solvent, the subcritical extraction method is the best for the extraction of total phthalocyanine lactones.
ABSTRACT
In the present study, a series of novel nitric oxide-hydrogen sulfide releasing derivatives of (S)-3-n-butylphthalide ((S)-NBP) were designed, synthesized, and evaluated as potential antiplatelet agents. Compound NOSH-NBP-5 displayed the strongest activity in inhibiting the arachidonic acid (AA)- and adenosine diphosphate (ADP)-induced platelet aggregation in vitro, with 3.8- and 7.0-fold more effectiveness than (S)-NBP, respectively. Furthermore, NOSH-NBP-5 could release moderate levels of NO and HS, which would be beneficial in improving cardiovascular and cerebral circulation. Moreover, NOSH-NBP-5 could release (S)-NBP when incubated with rat brain homogenate. In conclusion, these findings may provide new insights into the development of novel antiplatelet agents for the treatment of thrombosis-related ischemic stroke.