百可利对6-羟多巴胺不同注射位点帕金森病模型大鼠的治疗作用

目的观察百可利对6-羟多巴胺(6-OHDA)内侧前脑束(MFB)和纹状体尾壳核(CPu)两个不同注射位点帕金森病(PD)模型大鼠的治疗作用,两个注射位点模型分别记为:MFB-M,CPu-M。方法运用6-OHDA两点注射法,损毁大鼠左侧中脑多巴胺能神经元,制备PD模型。记录大鼠后肢肌电(EMG)信号频率观察肌肉震颤;测定大鼠自主活动;电化学法检测纹状体内多巴胺(DA)及其代谢产物含量;免疫组化法检测大鼠脑内酪氨酸羟化酶(TH)、OX-42表达;观察神经元超微结构变化。结果给药3周后,两个注射位点的模型组行为改变趋势一致,百可利在两个注射位点的模型动物上药效表现不同,在CPu-M组可明显提高PD大鼠自主活动数(P<0.05)。EMG信号分析显示,在MFB-M组,给予百可利,肌电频率降低55%;在CPu-M组,给予百可利,肌电频率降低60%。EMG时效研究表明,在CPu-M组,百可利药效持续420 min以上。纹状体递质水平显示,两个注射位点的模型组DA递质水平差异很大,在CPu-M组,百可利能够明显升高DA水平(P<0.05)。两个注射位点的模型组免疫组织化学结果趋势一致,在CPu-M组,百可利有更明显神经元保护作用(P<0.05),在MFB-M组,百可利抑制小胶质细胞过度激活作用更强(P<0.01)。结论不同注射位点制备的PD模型能够反映不同时期PD的病理变化,百可利可通过抑制炎性介质生成和释放、保护残存神经元、恢复神经元功能等机制改善PD不同发病时期模型动物的行为学症状。

牻牛儿苗的化学成分研究

目的:研究牻牛儿苗(Erodium stephanianum Willd.)的化学成分。方法:采用硅胶、SephadexLH-20柱层析并结合重结晶等方法对牻牛儿苗的95%乙醇提取物的乙酸乙酯萃取部位进行分离纯化,再依据理化性质和波谱数据(包括NMR和MS)对所分的化合物进行结构鉴定。结果:分离并鉴定了5个化合物,分别为:没食子酸(gallic acid,Ⅰ)、山奈酚(kaempferol,Ⅱ)、山奈酚-3-O-β-D-吡喃葡萄糖苷(kaempferol-3-O-β-D-glucopyranoside,Ⅲ)、柯里拉京(corilagin,Ⅳ)、鞣花酸(ellagicacid,Ⅴ)。结论:化合物Ⅲ为首次从该植物中发现。实验结果可为牻牛儿苗的进一步研究开发提供依据。

Hige namine as a Potential Pharmacologic Stress Age nt in the Detection of Coro nary Artery Disease

Myocardial perfusion imaging(MPI) is valuable for the diagnosis,prognosis,and management of coronary artery disease(CAD).The most commonly used pharmacologic stress agents at present are vasodilators and adrenergic agents.However,these agents have contraindications and may cause adverse effects in some patients.Thus,other stress agents feasible for more patients are required.Higenamine(HG) is a β-adrenergic receptor agonist currently approved for clinical trials as a stress agent for myocardial infarction.It also has a promising value in MPI for the detection of CAD in preclinical and clinical studies.This review summarizes the application of HG on MPI,including its mechanism of action,stress protocol,efficacy,and safety.

天然异黄酮类化合物的药理学研究进展

天然异黄酮类化合物广泛存在于植物界中,是许多中草药的有效成分,现已发现其多种药理学活性,受到广泛的关注。本文通过收集和整理近10年来关于天然异黄酮类化合物的研究文献,较全面地综述了其植物来源及药理作用,包括雌激素样作用,抗肿瘤作用,防治心血管疾病的作用,对骨代谢的作用,神经保护作用和提高认知能力的作用及其作用机制,对于天然异黄酮类化合物的开发与利用提供指导信息。

乙酰胆碱酯酶抑制剂高通量筛选方法的优化及应用

目的:优化乙酰胆碱酯酶抑制剂高通量筛选方法,筛选发现结构新颖的抗阿尔茨海默病药物活性化合物。方法:采用DTNB法,建立乙酰胆碱酯酶抑制剂筛选模型,考察了反应体系的最大吸收波长、底物浓度、酶浓度、pH值、孵育温度和时间对系统的影响,最终确定反应体系的最佳条件,并通过Z'因子分析,考察模型的稳定性。结果:酶反应体系的最佳条件为检测波长412 nm,底物浓度5 mmol.L-1,酶稀释倍数20倍,孵育温度37℃,孵育时间30 min,pH 7.4～8.0;测得该模型的Z'因子为0.654。通过乙酰胆碱酯酶抑制剂的高通量筛选,发现了一批乙酰胆碱酯酶抑制剂。结论:优化后的乙酰胆碱酯酶抑制剂高通量筛选模型稳定可靠,适于高通量筛选。

Cdk5/p35抑制剂高通量筛选方法的建立及应用

目的:建立适于高通量筛选的Cdk5/p35抑制剂筛选模型。方法:Cdk5/p35与其底物Histone H1反应时需消耗ATP,通过测定体系中ATP的剩余量,考察ATP浓度、酶浓度、底物浓度、反应温度、反应时间及体系pH等因素对酶促反应的影响,进而确定酶与底物反应的最佳条件,并采用Z'因子评价该模型的可靠性和可行性。应用已建立的Cdk5/p35抑制剂高通量筛选模型筛选和发现活性化合物。结果:反应体系的最佳反应条件为:ATP终浓度3μmol·L-1,Cdk5/p35终浓度30 nmol·L-1,Histone H1终浓度70μmol·L-1,孵育温度37℃,孵育时间60 min,体系pH 7,Z'-因子为0.66。应用该模型对222个虚拟筛选挑选出的化合物进行筛选,发现了13个对Cdk5/p35抑制活性较好的化合物。结论:实验结果表明,该模型具有稳定、灵敏、高效等特点,可用于Cdk5/p35抑制剂的高通量筛选。该模型的建立,为Cdk5/p35抑制剂的发现以及Cdk5/p35调控AD的机制研究奠定基础。

Xiao-Xu-Ming decoction extract alleviates LPS-induced neuroinflammation associated with down-regulating TLR4/MyD88 signaling pathway in vitro and in vivo

In the present study, we aimed to investigate the effects of Xiao-Xu-Ming decoction extract(XXM) on lipopolysaccaride(LPS)-induced neuroinflammation in vitro and in vivo. In vitro, the microglia BV2 cells were treated with 200 ng/mL LPS for 24 h to induce inflammatory responses. In vivo, mice were treated with 5 mg/kg LPS to induce inflammatory responses. The NO level was determined by Griess Reagents. The levels of IL-1β, IL-6, TNF-α and MCP-1 were determined by ELISA. The expressions of Iba-1, TLR4 and MyD88 at the protein levels were determined by Western blotting analysis. The mRNA levels of TLR4 and MyD88 were determined by real-time PCR. In vitro, XXM significantly reduced the levels of various pro-inflammatory factors, including NO, IL-1β, IL-6 and TNF-α, induced by LPS in the supernatant of BV2 cells and suppressed expressions of inflammatory proteins TLR4 and MyD88 induced by LPS in BV2 cells. In vivo, XXM significantly inhibited microglia activation, attenuated LPS-induced inflammatory factors and chemokine production, such as IL-1β, IL-6, TNF-α and MCP-1, and inhibited the expressions of inflammatory proteins including TLR4 and MyD88, in the cortex of LPS-induced mice. Our findings suggested that XXM could attenuate LPS-induced neuroinflammation via down-regulating TLR4/MyD88 signaling pathway.

Esculin alleviates acute kidney injury and inflammation induced by LPS in mice and its possible mechanism

Acute kidney injury(AKI)is a common clinical serious illness.Esculin(ES)is a coumarin compound of traditional Chinese medicine Cortex Fraxini.Our previous study has found that ES protects against inflammation and renal damage in diabetic rats.In the present study,we aimed to investigate the effects and the possible mechanism of ES against lipopolysaccharides(LPS)-induced AKI in mice.Renal morphology was observed by H&E staining.Renal function was evaluated by blood urea nitrogen(BUN)level and creatinine content in serum.Inflammatory factor levels were measured by ELISA assay.The inflammatory proteins were analyzed by RT-PCR and Western blotting analysis.The results showed that ES alleviated LPS-induced pathological injury and renal dysfunction,and decreased BUN level and creatinine content in serum.In addition,ES significantly reduced the release of pro-inflammatory factors,including IL-1β,IL-6 and TNF-α,chemokine MCP-1 and cell adhesion molecule ICAM-1.Furthermore,the expressions of inflammatory pathway proteins P2 X7,HMGB1,TLR4 and MyD88 both at the mRNA and protein levels were all down-regulated by ES in the kidney tissue of LPS-challenged mice.These results suggested ES protected against LPS-induced AKI through inhibiting P2 X7 expression and HMGB1/TLR4 inflammatory pathway.

Xiao-Xu-Ming decoction extract ameliorates brain injury in rats with thrombotic focal ischemic stroke and understanding possible therapeutic targets using proteomics

Ischemic stroke seriously threatens human health and quality of life.Xiao-Xu-Ming(XXM)decoction has been a classical prescription for stroke therapy.In our previous studies,we have found that XXM exerts neuroprotective effects,improves brain injury,and attenuates neuroinflammation in cerebral ischemia rats.In this study,we investigated the effects and possible mechanism of XXM on thrombotic focal cerebral ischemia.After treatment with XXM,the neurological function and motor abilities were improved,and cerebral infarction volume was significantly decreased compared with rats of thrombotic focal cerebral ischemia.Besides,the results of BBB integrity detected by EB leakage and tight junction(TJ)protein expression showed that XXM could maintain BBB integrity and improve the expressions of TJ proteins,including claudin-1,occluding,and ZO-1,in the ischemic ipsilateral cortex disrupted by thrombotic cerebral ischemia.Furthermore,proteomic techniques were used to identify the differentially expressed proteins(DEPs)in the ischemic cerebral cortex,and the results showed that 132 DEPs regulated by XXM were detected in the ischemic cerebral cortex.Bioinformatic analysis showed that these regulated proteins by XXM were mainly involved in complement and coagulation cascade,and lysosome,etc.Furthermore,there was an interaction among DEPs,including Lgals3,Ctsz,Capg,C1qa,S100a4,Grn,Hspb1,Aif1,and Anxa1,etc.In conclusion,XXM ameliorated brain injury of thrombotic focal ischemic stroke,and Lgals3,Ctsz,Capg,C1qa,S100a4,Grn,Hspb1,Aif1,and Anxa1 could help provide possible therapeutic targets of XXM for ischemic stroke and offer research direction for further research.

Network pharmacological analysis of Xiao-Xu-Ming decoction against ischemic stroke and verification of its mechanism of anti-inflammation and neurovascular protection in vivo

Stroke is a major cause of severe disability and death.Xiao-Xu-Ming decoction(XXMD)is an effective prescription for stroke and its sequelae,while its effective ingredients and mechanism are still unclear.In the present study,we aimed to explore the effective ingredients and mechanism of XXMD in treating cerebral ischemia using network pharmacology.The main chemical components and targets of 12 herbs of XXMD were obtained by the TCMSP database and analysis platform database.The active components in XXMD were screened according to oral utilization and drug-like properties.Then,the cerebral ischemia targets were obtained through GeneCards,OMIM,TTD,Diligent and Drugbank databases.We analyzed the pathophysiological processes and pathways involved in the treatment of cerebral ischemia with XXMD by using the Metascape data analysis platform.Results showed thatβ-sitosterol,kaempferol,quercetin,stigmasterol,wogonin,and catechins might be the potential core active ingredients of XXMD in the treatment of cerebral ischemia.The therapeutic effect of XXMD on stroke was mainly exerted through regulating neuroinflammatory response and neurovascular protection.Furthermore,the anti-neuroinflammation and neurovascular protection of XXMD were further confirmed using cerebral ischemia rats.Collectively,our findings revealed that the mechanism of XXMD on the treatment of cerebral ischemia was related to anti-neuroinflammation and neurovascular protection.

Rapid and sensitive HPLC-MS/MS method for quantitative determination of isochlorogenic acid B in rat plasma and its application in pharmacokinetic study

A sensitive LC-ESI-MS/MS method for determination of isochlorogenic acid B in rat plasma was developed and validated in the present study. Plasma samples were prepared by a simple protein precipitation with methanol containing resveratrol as internal standard (IS). The chromatographic separation was performed on a Zorbax SB-Cjg column (3.5 pm, 2.1 mmx 100 mm, Agilent, USA) at a flow rate of 0.2 mL/min using methanol/water containing 0.1% formic acid (v/v) as mobile phase. The detection was performed on a triple quadrupole tandem mass spectrometer equipped with Electronic Spray Ion by selected reaction monitoring (SRM) of the transitions at m/z 515.3->352.9 for isochlorogenic acid B and m/z 227.1-143.1 for IS, respectively. The calibration curve of the method was linear over the range of 5-2500 ng/mL (r^2= 0.9982). The intra- and inter-day precisions (R.S.D.%) were less than 12.46%, and the accuracy (R.E.%) was within ±5.80%. Isochlorogenic acid B was sufficiently stable under all relevant analytical conditions. The validated method was successfully applied to the plasma pharmacokinetic studies of isochlorogenic acid B in rats. It was found that isochlorogenic acid B had non-linear pharmacokinetic characteristics in rats within the dosage ranges from 5 to 20 mg/kg.

Determination of deferasirox particle size distribution via laser diffraction and its application in establishing a correlation between particle size and drug dissolution in vitro

Deferasirox is the first-line drug for iron overload due to thalassemia in adults and pediatric patients. It is classified as a type II compound in the Biopharmaceutics Classification System, and thus the particle size of its active pharmaceutical ingredient(API) should be strictly controlled during the manufacturing process. In the present study, laser diffraction was adopted to measure the particle size distribution of deferasirox API. We also developed and validated an accurate and convenient method by investigating important optical parameters and sample dispersing conditions. The relative standard deviation values, namely, d(0.1), d(0.5), d(0.9), and d(4,3), measured via methodology validation and actual sample measurement were < 3%. The dissolution curves of several batches of dispersible tablets prepared using deferasirox with different particle sizes were compared in the four dissolved media to investigate the influence of particle size on drug dissolution in vitro. Results indicated that the particle size distribution of deferasirox API significantly affected the release of its dispersible tablet.

The synthesis and antibacterial activity evaluation of oxazolidinone-deferasirox conjugates

We reported herein the synthesis and antibacterial activity evaluation of two oxazolidinone-deferasirox conjugates with different linkers that were designed based on the“Trojan horse”strategy.The conjugates could combine with Fe^(3+)ions as the deferasirox.However,both conjugates were inactive against tested bacteria,including S.aureus,E.coli,A.baumannii,and P.aeruginosa.The results suggested that the synthesized iron chelator deferasirox was not suitable as a siderophore of the bacteria to transport the antibiotic,or the coupling linker of the synthesized conjugates could not be hydrolyzed to release the oxazolidinone in the cytoplasm.Therefore,the design and synthesis of oxazolidinone-deferasirox conjugates need further exploration.

Biocatalytic bis-C-alkylation of phenolics using one-pot cascades with promiscuous C-glycosyltransferase and prenyltransferase

C-glycosylation and C-prenylation are two important C-C-bond forming reactions for preparation,diversification and structural modification of natural/unnatural products with pharmacological activities.Here,we described unprecedented enzymatic cascades to C-glycosylate/prenylate different acyl resorcinol derivatives in stepwise,one-pot reactions by combining two promiscuous enzymes,MiCGT,a C-glycosyltransferase,and AtaPT,a prenyltransferase.Five novel bis-C-alkylated products were obtained and structurally identified by MS and NMR spectroscopic data as well as comparison with the literature.This study provided a potential synthetic strategy for synthesizing structurally novel and diverse compounds bearing both C-glycosyl and C-prenyl moieties by a two-step,enzymatic bis-C-alkylation.

Three new compounds from endophytic fungus Periconia sp.F-31

Three new compounds(1–3), including a chlorine-containing dihydroisocoumarin pericochlorosin A(1), a chlorinated phenol pericochlorosin B(2) and a decalin derivative pericoannosin G(3), were isolated from endophytic fungus Periconia sp. F-31 of the medicinal plant Annona muricata. The structures and absolute configurations were elucidated by extensive spectroscopic methods and calculated electronic circular dichroism analysis. Compound 2 displayed potent anti-HIV activity with IC50 value of 2.2 μM.