血塞通与阿司匹林联合用药对大鼠脑缺血再灌注损伤的保护作用

Objective:To investigate the protective effects of the combination of Xuesaitong(XST)and aspirin on cerebral ischemia and reperfusion injury(CIRI)in rats,and further explore the underlying mechanisms.Methods:A total of 150 male Sprague-Dawley(SD)rats were randomly divided into five groups with 30rats in each group:sham group,middle cerebral artery occlusion/reperfusion(MCAO/R)model group,XST group,aspirin group,and XST+aspirin group.Rats were pretreated with XST,aspirin,or XST+aspirin for7 d.One hour after the last administration,a model of CIRI was induced by MCAO/R.Neurological deficits were assessed using Longa’s five-point scale.Cerebral edema was detected by the measurement of brain water content.The volume of cerebral infarction was determined by 2,3,5-triphenyltetrazolium chloride(TTC)staining.The activities of superoxide dismutase(SOD),catalase(CAT),and glutathione peroxidase(GSH-Px),as well as levels of malonaldehyde(MDA)were detected by commercial kits.Enzyme-linked immunosorbent assay(ELISA)was used to determine the levels of interleukin-1(),interleukin-4(IL-4),interleukin-6(IL-6),interleukin-10(IL-10),tumor necrosis factor-alphamonocyte chemotactic protein 1(MCP-1),and kynurenine in serum,cerebral cortex,and hippocampus of MCAO/R rats.The protein expression of nuclear factor erythroid 2-related factor(Nrf2),heme oxygenase-1(HO-1),I-kappa B alpha(IκBα),and nuclear factor kappa B(B)/p65 in the cortex were analyzed by western blotting.Results:Treatment of XST,aspirin,and XST+aspirin significantly alleviated the neurological deficits,cerebral edema,and cerebral infarct volume induced by MCAO/R.Treatment of XST,aspirin,and XST+aspirin also reduced MDA,,MCP-1,and kynurenine levels,and increased SOD,CAT,GSH-Px,IL-4,and IL-10 levels in serum,cerebral cortex,and hippocampus of MCAO/R rats.Furthermore,treatment of XST,aspirin,and XST+aspirin decreased the expression of nuclearB/p65 and increased the expression of IκBα,nuclear Nrf2,and HO-1.Importantly,the combination of XST and aspirin enhanced the protective effects of XST or aspirin treatment alone on CIRI in rats.Conclusion:The combination of XST and aspirin significantly inhibited oxidative stress and inflammation in serum,cerebral cortex,and hippocampus of MCAO/R rats.The combination of XST and aspirin exerted more protective effects than XST or aspirin treatment alone.The combination of XST and aspirin might provide the synergistic therapeutic effects on CIRI,and deserve further clinical investigation.     

Tectorigenin inhibits the inflammation of LPS-induced acute lung injury in mice

AIM： In a previous study, the anti-inflammatory effects of tectorigenin were disclosed. In this study, the anti-inflammatory effects of tectorigenin on acute lung injury using a lipopolysaccharide（LPS）-induced acute lung injury（ALI） mouse model were investigated. METHOD： The cell-count in the bronchoalveolar lavage fluid（BALF） was measured. The animal lung edema degree was evaluated by the wet/dry weight（W/D） ratio. The superoxidase dismutase（SOD） activity and myeloperoxidase（MPO） activity was assayed using SOD and MPO kits, respectively. The levels of inflammatory mediators, including tumor necrosis factor-α（TNF-α）, IL-1β, and IL-6 were assayed using an enzyme-linked immunosorbent assay method. Pathological changes of lung tissues were observed through HE staining. The inflammatory signal pathway related protein nuclear factor NF-κB p65 mR NA expression was measured by real-time PCR, and the protein level of NF-κB p65 was measured using Western blotting analysis. RESULTS： The data showed that treatment with the tectorigenin markedly attenuated the inflammatory cell numbers in the BALF, decreased nuclear factor NF-κB p65 mR NA level and protein level in the lungs, and improved SOD activity and inhibited MPO activity. Histological studies showed that tectorigenin substantially inhibited LPS-induced neutrophils in lung tissue compared with the model group. CONCLUSION： The results indicated that tectorigenin had a protective effect on LPS-induced ALI in mice.     

熄风康对大鼠脑缺血保护作用▲

The protective effects of Xi Feng Kang (XFK) were studied in cerebral ischimia rats induced by ligaturing right common carotid artery. The results indicated that XFK could significantly reduce edema of brain tissue cells,enhance tPA and AT-Ⅲ activity,decrease PAI,vWF and RBC aggregation index of cerebral ischimia rats,significantly protect the experimental cerebral ischimia rats. The study prompted that the drug has the function of preventing cerebral thromloosis and reducing the injury of cerebral ischimia.     

Combination of Shuxuetong and aspirin protects against cerebral ischemia/reperfusion injury through ameliorating coagulation and fibrinolysis system

Objective:To explore the influence of the combination of Shuxuetong(SXT)and aspirin on coagulation and fibrinolytic system of rats.Methods:Suture method was applied to establish focal cerebral ischemia-reperfusion injury models in rats.SD rats were randomly divided into sham group,middle cerebral artery occlusion/reperfusion(MCAO/R)group,aspirin group,SXT group,and SXT+aspirin group(SA).The neurological deficits were assessed according to Longa’s grade 5 scoring method.The cerebral edema was detected by measuring the content of water in brain tissue.The volume of cerebral infarction was observed by 2,3,5-Triphenyltetrazolium chloride(TTC)staining.Blood plasma was collected by abdominal aortic method to test maximum platelet aggregation rate and four blood coagulation.CD61,CD62p,6-keto prostaglandin F1αantithrombinⅢ(AT-Ⅲ),D-dimer,plasminogen activator inhibitor-1(PAI-1),tissue factor(TF),tissue plasminogen activator(t-PA),platelet thromboxaneand von Willebrand factor(v WF)content in rat plasma were detected by ELISA.Results:SXT combined with aspirin could improve the neurological deficits,alleviate cerebral edema,and decrease the cerebral infarct value.Compared with the sham operation group,fibrinogen(FIB),6-AT-III,and t-PA in model group were significantly decreased;Compared with the model group,the above-mentioned indexes in SXT and aspirin treatment group were significantly increased.The prothrombin time(PT),activated partial thromboplastin time(APTT),thrombin time(TT),D-dimer,PAI-1,TF,TXB2,and v WF of the model group were significantly increased;The above-mentioned indexes in blood SXT+aspirin treated group were significantly decreased.There was a significant difference between the combined group and SXT group.The maximum concentration of plateletsin aspirin treated rats was significantly decreased,however,MPAR was reversed in SXT+aspirin treated group.Conclusion:SXT combined with aspirin can effectively inhibit platelet activation,regulate the maximum concentration of platelets,and improve coagulation function and fibrinolysis system.     

Astragaloside IV prevents lipopolysaccharide-induced injury in H9C2 cardiomyocytes

This study aimed to investigate the protective effects of astragaloside IV(AS IV) on lipopolysaccharide(LPS)-induced injury in H9C2 cardiomyocytes. H9C2 Cardiomyocytes were cultured with LPS(10 μg·mL-1) for 4 h and treated with AS IV at 50, 100, and 150 μmol·L-1 for various durations. Cell viability was determined by MTT. The content of released TNF-α and IL-6 from cardiomyocytes were evaluated by enzyme-linked immunosorbent assay(ELISA). The levels of superoxidase dismutase(SOD), malondialdehyde(MDA), lactate dehydrogenase(LDH), and creatine phosphate kinase(CK) were measured by using commercial available kits. The mR NA and protein expression levels of NF-κB p65 were measured by RT-PCR and Western blotting, respectively. And the NF-κB p65 activity was measured by ELISA. Our results demonstrated that AS IV at 50, 100, and 150 μmol·L-1 markedly inhibited the release of TNF-α and IL-6 and decreased NF-κB expression, compared with the model group. Moreover, the improved SOD activity and decreased MDA, LDH and CK levels were detected after AS IV treatment. In summary, AS IV could increase the activities of antioxidant enzymes, inhibite lipid peroxidation, and down-regulate the inflammatory mediators involved in the inflammatory responses. These results demonstrated that AS IV could prevent LPS-induced injury in cardiomyocytes.     

Anti-inflammatory and hepatoprotective effects of total flavonoid C-glycosides from Abrus mollis extracts

The aim of this study was to evaluate the anti-inflammatory and hepatoprotective effects of the total flavonoid C-glycosides isolated from Abrus mollis extracts（AME）. In the anti-inflammatory tests, xylene-induced ear edema model in mice and carrageenan-induced paw edema model in rats were applied. The hepatoprotective effects of AME were evaluated with various in vivo models of acute and chronic liver injury, including carbon tetrachloride（CCl4）-induced hepatitis in mice, D-galactosamine（D-GalN）-induced hepatitis in rats, as well as CCl4-induced hepatic fibrosis in rats. In the acute inflammation experiment, AME significantly suppressed xylene-induced ear edema and carrageenan-induced paw edema, respectively. In the acute hepatitis tests, AME significantly attenuated the excessive release of ALT and AST induced by CCl4 and D-GalN. In CCl4-induced hepatic fibrosis model, AME alleviated liver injury induced by CCl4 shown by histopathological sections of livers and improved liver function as indicated by decreased liver index, serum ALT, AST, TBIL, and ALP levels and hydroxyproline contents in liver tissues, and increased serum ALB and GLU levels. These results indicated that AME possesses potent anti-inflammatory activity in acute inflammation models and hepatoprotective activity in both acute and chronic liver injury models. In conclusion, AME is a potential anti-inflammatory and hepatoprotective agent and a viable candidate for treating inflammation, hepatitis, and hepatic fibrosis.     

Shenqi Xingnao Granules ameliorates cognitive impairments and Alzheimer’s disease-like pathologies in APP/PS1 mouse model

Objective: Alzheimer''s disease?(AD) is along with cognitive decline due to amyloid-β (Aβ) plaques, tau hyperphosphorylation, and neuron loss.?Shenqi Xingnao Granules (SQXN), a traditional Chinese medicine, significantly ameliorated the cognitive function and daily living abilities of patients with AD. However, till date, no study has investigated the mechanism of action of SQXN on AD. The present study aimed to verify the effects of SQXN treatment on cognitive impairments and AD-like pathologies in APP/PS1 mice. Methods: Four-month-old?APP/PS1?transgenic (Tg) mice were randomly divided into a model group and SQXN-treated (3.5, 7, 14 g/kg per day) groups. Learning-memory abilities were determined by Morris water maze and object recognition test. All mice were sacrificed and the brain samples were collected after 75 d. The soluble Aβ contents were detected by Elisa kit; The levels of expression of NeuN, APP, phosphorylated tau and related protein were measured by Western blotting; The inflammation factors were detected by the proinflammatory panel kit. Results: Four-month-old?APP/PS1?mice were administered SQXN by oral gavage for 2.5 months. Using the Morris water maze tests and Novel object recognition, we found that SQXN restored behavioral deficits in the experimental group of Tg mice when compared with the controls. SQXN also inhibited neuronal loss?(NeuN?marker). SQXN treatment decreased soluble Aβ42 through inhibiting the expression of sAPPβ and BACE-1 without regulating full-length amyloid precursor protein (FL APP). Insulin degrading enzyme (IDE), the Aβ degrading enzyme, were increased by SQXN. In addition, SQXN reduced hyperphosphorylated tau protein levels and prevented excessive activation of p-GSK-3β in the brain of APP/PS1 mice. Compared with APP/PS1 transgenic negative mice, IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-12p70, KC/GRO and TNF-α were not obviously changed in the brain of 6.5-month-old APP/PS1?transgenic (Tg) mice. However, SQXN could inhibited the expression of IL-2. Conclusion: These results demonstrate that SQXN ameliorates the cognitive impairments in APP/PS1 mice. The possible mechanisms involve its inhibition of neuronal loss, soluble Aβ deposition, tau hyperphosphorylation and inflammation.     

Pharmacology and Toxicology of Extract from Arcangelisia gusanlung

Objective To study the pharmacology and toxicology of the extracts from Arcangelisia gusanlung(EAG). Methods The anti-inflammatory activities were investigated using various inflammatory models including ear edema induced by xylene in mice, paw edema induced by carrageenan, and cotton pellet granuloma in rats. The analgesic effect was observed in hot-plate test and writhing test in mice and the antipyretic effect was observed in rat fever model induced by yeast. The antitussive action was tested in mice by sequential method and expectorant action was evaluated by tracheal excretion of phenol red. The antidiarrhea function was observed on normal intestinal propulsion of mouse model of diarrhea induced by decoction of Sennae Folium. The toxicity was measured by toxicological experiment.Results Each dose of EAG could significantly inhibit the paw edema, cotton pellet granuloma, and intestinal propulsion. EAG significantly reduced writhing times and amount of wet manure. Obvious antipyretic action to fevered rat was observed. EAG obviously increased the tracheal excretion of phenol red and prolonged the latency of cough. No toxic reaction was shown in the observed period, and the maximum tolerance dose of mice was equivalent to 1360 times of common-used dose in human. Conclusion The clinical dosage of EAG is safe, and its anti-inflammatory, analgesia, antipyresis, antitussive, expectorant, and antidiarrhea effects are significant.     

Metabolite Characterization of Penta-O-galloyl-??￠-D-glucose in Rat Biofluids by HPLC/QT of MS

Objective: To understand the in vivo metabolic fate of 1,2,3,4,6-Penta-O-galloyl-β-d-glucose(PGG)naturally existed in many medicinal herbs and food plants such as Rhus chinensis,Paeonia suffruticosa,Paeonia lactiflora and Mango.Methods: The metabolites of PGG in rat biofluids were characterized using high performance liquid chromatography combined with quadrupole time-of-flight tandem mass spectrometry(HPLC-QTOF-MS).Results: Ten metabolites in urine,five metabolites in feces and two metabolites in plasma,were observed when the rats were administrated with a single intravenous injection of PGG(20 mg/kg).Conclusion: PGG is firstly metabolized to gallic acid,then gallic acid undergoes sulfation,glucuronidation and methylation by rat liver.The determination of metabolites and the proposed metabolic pathway of PGG in vivo will be benefit to gain deeper insights into its pharmacological activities.     

Acupuncture at Kŏngzuì (孔最LU 6) for 76 cases of hemorrhoids

Objective

To observe the effect of acupuncture at K?ngzuì (孔最LU 6) for hemorrhoids.

Ergosta-7,22-diene-2β,3α,9α-triol(EGDT) from Ganoderma lucidum inhibits nasopharyngeal carcinoma cells by blocking EGFR signaling pathway

Objective

To investigate the efficacy and mechanism of EGDT against NPC cell lines.

疏血通与阿司匹林合用能够改善脑缺血再灌注损伤后凝血纤溶系统

目的：观察疏血通(SXT)与阿司匹林合用对脑缺血再灌注损伤大鼠凝血及纤溶系统的影响。 方法：选用经典的线栓法制备大鼠局灶性脑缺血再灌注损伤模型。将50只雄性SD大鼠随机分为假手术组、脑中动脉栓塞（MCAO/R）模型组、阿司匹林(10 mg/kg)组、疏血通(0.6 ml/kg)组、疏血通(0.6 mg/kg)+阿司匹林(10 mg/kg)组。按照Longa的5级评分方法进行神经功能缺损评分;干湿重法测定脑组织含水量;四氮唑红（TTC）染色测定脑梗塞体积;腹主动脉取血检测血小板聚集率（MPAR）、凝血四项并用酶联免疫法检测大鼠血浆中血小板CD61、CD62p、 6-酮前列腺素F1α(6-keto-PGF1α)、抗凝血酶Ⅲ(AT-Ⅲ)、D-二聚体(D-dimer)、纤溶酶原激活物抑制剂-1(PAI-1)、组织因子 (TF)、组织型纤溶酶原激活物(t-PA)、血小板血栓烷素B2 (TXB2)、血管性血友病因子（vWF）的含量。 结果：疏血通与阿司匹林合用能够改善MCAO/R诱导大鼠神经功能缺损,减轻脑水肿,缩小脑梗死体积。与假手术组相比较纤维蛋白原（FIB）、6-PGF1α、AT-Ⅲ、t-PA均有显著的下降;与模型组相比较,疏血通与阿司匹林合用组的上述指标都有明显上调。模型组的凝血酶原时间（PT）、活化部分凝血活酶时间（APTT）、凝血酶时间（TT）、CD61、CD62p、D-dimer、PAI-1、TF、TXB2和vWF与假手术组相比都有明显上升;经疏血通与阿司匹林处理组的上述指标都有明显下降。合用效果与单用药比较,差异具有显著性。与假手术组比较,MPAR显著升高;阿司匹林治疗后,MPAR显著下调;疏血通与阿司匹林合用给药组MPAR下调水平降低。 结论：疏血通与阿司匹林合用能抑制血小板活化,调节血小板最大聚集率,改善凝血功能和纤溶系统。     

赤芍通过抗神经胶质增生及抗氧化保护局部短暂性脑缺血小鼠海马部位神经的研究

目的：卒中是全球第二大死亡原因。本研究探究赤芍（Paeoniae Radix Rubra， PRR）对缺血性脑卒中小鼠的神经保护作用。方法：使用小鼠大脑中动脉堵塞法构建局部脑缺血模型。将实验动物分为4组，假手术组（Sham 组）、假手术给药组（PRR-sham组）、手术组（Isch组）、手术给药组（PRR-Isch组）。采用TTC染色法测量各组小鼠脑梗死体积；免疫组织化学染色观察各组之间小鼠缺血侧大脑海马区神经元细胞的凋亡情况以及星形胶质细胞和小胶质细胞的活化程度；采用蛋白印迹法观察小鼠缺血侧海马超氧化物歧化酶1（SOD1）、超氧化物歧化酶2（SOD2）、过氧化氢酶抗体（Catalase）表达水平。结果：赤芍可以明显减少局部脑缺血小鼠的脑梗死体积，并能抑制海马CA1区星形胶质细胞和小胶质细胞的活化程度，同时可以改善由缺血再灌注损伤引起的SOD1、SOD2和Catalase的表达水平下降情况。结论：赤芍可以通过抗胶质增生以及抗氧化作用改善局部短暂性脑缺血小鼠的神经损伤情况。     

人参水煎液miRNA的提取及验证

目的：证明新鲜人参汤存在微小RNA（miRNA）。方法：按照传统方法将新鲜人参煎煮成汤剂,浓缩,用植物microRNA提取试剂盒提取miRNA。然后用DNase I处理miRNA,进行琼脂糖凝胶电泳和Agilent 2100生物分析。选取在人参中高表达的MiR-159和miR-6135进行实时定量PCR验证。结果： MiR-159和miR-6135在新鲜水煎液中表达,水平低于鲜人参。结论：MiRNA在加工后稳定存在,高温处理后保持稳定。这些发现为进一步研究人参miRNAs奠定了基础。     

紫杉醇和鸦胆子油分子配型纳米乳的制备及抗肿瘤作用的体内外评价

目的 制备共同时递送紫杉醇-油酸前药(PTX-OA)和鸦胆子油(BJO)的分子配型纳米乳制剂(CMNEs)。方法 对基于分子配型组装技术的紫杉醇-油酸/鸦胆子油纳米乳制剂体外对HepG2细胞的抑制、细胞周期、细胞凋亡和体内抑制裸鼠肿瘤生长的评价。结果 紫杉醇-油酸/鸦胆子油纳米乳（PTX-OA/BJO CMNEs）粒径为108.7 2.3 nm,包封率> 95%。PTX-OA/BJO CMNEs对HepG-2细胞的毒性作用有浓度和时间依赖性,细胞周期试验表明PTX-OA/BJO CMNEs增加了G2/M期阻滞。注射PTX-OA/BJO CMNEs后的裸鼠模型中,对比注射生理盐水组,裸鼠肿瘤体积明显减少 (P<0.05),表明PTX-OA/BJO CMNEs在体内有很好抗肿瘤效果。进一步研究发现,PTX-OA/BJO CMNEs的抗肿瘤作用增强与诱导肿瘤细胞凋亡的能力有关,尤其是PTX-OA/BJO CMNEs可明显抑制肿瘤细胞的增殖和TOPO II的活性。结论 不同机制的两种药物联合给药能同时阻断不同的抗癌途径,从而提高治疗反应,降低毒性。     

Study on ``liver-soothing and mind-regulating'' acupuncture manipulation in regulating the blood-oxygen concentration in cerebral cortex of PTSD rats

Objective

To explore the brain function mechanism of "liver-soothing and mind-regulating" acupuncture manipulation in intervening post-traumatic stress disorder (PTSD).

Synthesis and Biological Evaluation of New Peroxo-bridged Diosgenin Derivatives

Objective: In order to find lead compound with anti-HBV activity from peroxo-bridged diosgenin derivatives obtained with Eosin Y as the photosensitizer.Method: Eosin Y was used as the photosensitizer to activate the oxygen in the air to synthesize novel diosgenin derivatives with peroxo-bridge.The structures of synthesized compounds were identified by NMR and HR-MS.Their cytotoxicity and antihepatitis B activity were evaluated via MTS assay and ELISA method,respectively.Results: Six diosgenin derivatives were synthesized,three of which contained peroxo-bridge,and their structures were confirmed by spectroscopy.It showed that 5 α,8 α-peroxo-6-alkenyl-diosgenin(7)could suppress the production of HBs Ag on transfected HepG2.2.15 cells at low-toxic concentration and the inhibition rate on HepG2.2.15 cells was 18.28% at 12.50 μg/mL,better than that of 3 TC(7.30% at 12.50 μg/mL)and others.Conclusion: Due to its lower cytotoxicity and potential anti-hepatitis B activity,compound 7 could be developed as the promising candidate of anti-hepatitis B drug.It also indicated that the peroxo-bridged derivatives had potential biological values for developing clinical agents.