The Patterns of Secretory Structure and Their Relation to Hypericin Content in Hypericum

利用整体透明、石蜡制片和半薄切片法 ,对金丝桃属 (HypericumL .) 8组 2 0种 1变种植物的分泌结构进行了比较解剖研究 ,结果表明 :该属植物的分泌结构可分为分泌细胞团和分泌囊两种类型。但在不同植物种和不同器官内 ,分泌结构的类型和分布密度存在差异。对上述植物的提取物进行薄层层析和高效液相层析检测 ,结果表明 :具有分泌细胞团的植物器官含有金丝桃素 ;而无分泌细胞团的植物器官 ,则不含金丝桃素 ,从而证明金丝桃素由分泌细胞团合成和贮藏。在前者中 ,其金丝桃素的含量与其分泌细胞团密度成正相关。

Differential diagnosis of gallstones by using hypericin as a fluorescent optical imaging agent

AIM: To explore the feasibility of using hypericin as an optical imaging probe with affinity for cholesterol for differential fluorescent detection of human gallstones.METHODS: Cholesterol, mixed and pigment stones from cholecystectomy patients were incubated with hypericin or solvent. After 72 h, the stones were analysed for fluorescence(365 nm) and treated with 2-propanol/dimethyl sulfoxide for high performance liquid chromatography(HPLC) analysis. Rats with virtual gallbladder containing human cholesterol, mixed or pigment gallstones(VGHG) received 5 mg/kg hypericin or solvent and VGHG rats with cholesterol stones were given different hypericin doses(5-15 mg/kg). Twelve hours later, the stones were analysed at 365 nm. Biliary excretion and metabolites of hypericin were assessed in common bile duct(CBD) cannulated rats for 9 h using fluorospectrometry, HPLC and matrixassisted laser desorption/ionization-time-of-flight mass spectrometry(MALDI-TOF MS).RESULTS: Homogeneous high fluorescence was seen on cholesterol stones either pre-incubated with hypericin or extracted from VGHG rats receiving hypericin. Mixed stones showed a dotted fluorescent pattern, whereas pigment and solvent-treated ones lacked fluorescence. HPLC showed 7.68, 6.65 and 0.08 × 10^(-3) M of cholesterol in extracts from cholesterol, mixed, and pigment gallstones, respectively. Hypericin accounted for 2.0, 0.5 and 0.2 × 10-6 M in that order. On cholesterol stones from VGHG rats receiving different hypericin doses, a positive correlation was observed between dose and fluorescence. In the bile from CBD-cannulated rats, fluorescence represented 20% of the injected dose with two peaks in 9 h. HPLC analysis revealed that hypericin conjugates reached 60% of the peak area. By MALDI-TOF MS, hypericinglucuronide was detected. CONCLUSION: This study proves the potential use of hypericin for differential fluorescent detection of human gallstones regarding their chemical composition.

Evaluation of photodynamic efficiency of an endogenous enzyme-NQO1 activated hypericin

OBJECTIVE To investigate the therapeutic efficiency of an endogenous enzyme-NQO1 activated hypericin in photodynamic therapy.METHODS An endogenous enzyme-NQO1 responsive photo sensitizer was designed and synthesized by conjugating aquinonebased ligand to a natural photosensitizer derived from Chinese herb.The photophysical and photochemical properties were investigated through UV-visible and fluorescence spectrophotometer,and the photodynamic activity was evaluated with MTT assay.RESULTS An endogenous enzyme-NQO1 activated hypericin was prepared and fully characterized with various spectroscopic methods.The electronic absorption was almost the same with the free hypericin,indicating the introducing of the ligand to hypericin has little effect to its ground state,while there is almost no detected fluorescence and reactive oxygen species(ROS)generation in PBS solution indicating the introducing of the ligand can effectively quench the fluorescence emission and ROS generation.The in vitro study showed that both compounds have almost no dark toxicity,but they are highlyphotocytoxic with an IC50 less than 1μmol·L-1 against A549 cell lines indicating the modified compound can be activated in the intracellular environment.CONCLUSION A simple and efficient hypericin-based activated photosensitizer was prepared.The ROS generation was quenched in PBS solution and it would be activated inside A549 cell lines.It may be served as apromising tumor selective fluorescent probe and photosensitizer for targeted photodynamic therapy.

Topically applied hypericin exhibits skin penetrability on nude mice

OBJECTIVE Hypericin,a powerful naturally photosensitizer in photodynamic therapy(PDT),is suitable for treating skin diseases involving excess capillary proliferation.In the present study,we aimed to evaluate the skin penetrability of a topically applied hypericin,expecting reducing the risk of prolonged skin photosensitivity,which often occurs after systemic administration.METHODS The Franz diffusion cell assay was performed to evaluate different penetration enhancers.In vivo studies,fluorescence microscopy was performed to examine the distribution of hypericin in the skin,macroscopic and microscopic analyses were also carried out to detect pathological changes in the skin after topical hypericin-PDT treatment.Immunohistochemistry was used to determine the expression of PECAM-1 in the treated skin.RESULTS 5% menthol facilitated hypericin penetrate the skin of nude mice most.The results of in vivo assays revealed that hypericin penetrated nude mice skin,spread to the dermis,and resulted in obvious photosensitivity reaction on the dermal capillaries.Moreover,skin injured by the photosensitive reaction induced by hypericin was replaced by normal skin 7 d after hypericin-PDT treatment.CONCLUSION Topical hypericin could penetrate nude mouse skin well and be great potential in PDT treatment of skin diseases.

In vitro 3D malignant melanoma model for the evaluation of hypericin-loaded oil-in-water microemulsion in photodynamic therapy

Advances in biomimetic three-dimensional(3D) melanoma models have brought new prospects of drug screening and disease modeling, since their physiological relevancy for recapitulating in vivo tumor architectures is more accurate than traditional two-dimensional(2D) cell culture. Gelatin methacryloyl(GelMA) is widely used as a tissue-engineered scaffold hydrogel for 3D cell culture. In the present study, an in vitro 3D malignant melanoma model based on Gel MA was fabricated to evaluate the efficiency of hypericin(Hy)-loaded microemulsion(ME) in photodynamic therapy against melanoma. The ME was produced by the spontaneous emulsification method to enhance the bioavailability of Hy at tumor sites. Hy-loaded MEs were applied to a 3D malignant melanoma model made using 6% Gel MA and the co-culture of B16F10 and Balb/c 3T3 cells,followed by crosslinking using violet light(403 nm). The observation revealed excellent cell viability and the presence of F-actin cytoskeleton network. Hy-loaded MEs exhibited higher phototoxicity and cell accumulation(about threefold) than free Hy, and the cells cultured in the 3D system displayed lower susceptibility(about 2.5-fold) than those in 2D culture.These findings indicate that the developed MEs are potential delivery carriers for Hy;furthermore, Gel MA hydrogel-based modeling in polydimethylsiloxane(PDMS) molds is a user-friendly and cost-effective in vitro platform to investigate drug penetration and provide a basis for evaluating nanocarrier efficiency for skin cancer and other skin-related diseases.

Effects of Different Geographical Aspects and Ontogenetic Variability on Total Hypericin Content of Hypericum triquetrifolium Turra. and Hypericum scabrum L.

The present study was conducted to determine the total hypericin contents of Hypericum triquetrifolium Turra.and Hypericum scabrum L.species which are naturally distributed in the flora of Siirt province,Turkey.Hypericin contents of Hypericum species grown in different geographical aspects(North,South,East,and West),and it was measured at different harvest times(full bloom-ing and post blooming period).In the current study,it has been determined that total hypericin content varies considerably according to aspects,plant developmental stages(ontogenetic variance),and species.According to species x aspect interaction,the highest total hypericin content was recorded from the west aspect(3.13 mg/g)in Hypericum triquetrifolium,while,the lowest hypericin content was also obtained from the west aspect(1.22 mg/g)in Hypericum scabrum.When the highest total hypericin content was analyzed according to aspect x species x harvest time interaction,the highest total hypericin content was produced from Hypericum triquetrifolium at the harvest of west aspect with 5.28 mg/g,while the minimum amount of hypericin was obtained from the same aspect in Hypericum scabrum with 0.50 mg/g.In species x harvest time interaction,the highest total hypericin content was obtained from the full bloom(3.10 mg/g)harvest in Hypericum triquetrifolium,while the lowest hypericin was obtained from the full bloom(1.26 mg/g)harvest in Hypericum scabrum.The data suggest that the aver-age total hypericin content was 2.26 mg/g in Hypericum triquetrifolium and 1.28 mg/g in Hypericum scabrum.

Effect of Photoactivated Hypericin on Growth and Antibiotic Susceptibility of Hospital-Related Staphylococcus aureus and Enterococcus sp. Clinical Strains

Resistance against commonly used antibiotics is a serious clinical problem in recent medical practice. There exist several bacterial strains in which the possibilities of their inhibition are very limited due to multidrug resistance. Antimicrobial photodynamic therapy (aPDT) represents an option how to effectively suppress the growth of resistant pathogens. In this work we have studied interactions of potent photosensitizer hypericin (Hyp) with hospital-related gram positive (Gram+) and gram negative (Gram-) bacterial strains and the effects of photodynamic activated Hyp on bacterial susceptibility and/or resistance of these strains to antibiotics. We demonstrated a significant influence of photoactivated Hyp on growth of Staphylococcus aureus and Enterococcus sp. We have also shown that it is extremely important to use the effective concentrations of Hyp for aPDT, which completely inhibit the growth of microorganisms. Otherwise, there appears an increase in resistance, probably due to the activation of efflux mechanisms, which are involved in the efflux of Hyp and antibiotics as well.

Effect of Hypericin on Confocal Imaging of Ca^(2+) Signaling in Cultured Human Retinal Pigment Epithelium

Purpose: To investigate the mechanism of the Ca2 + signaling in cultured human retinal pigment epithelial(RPE) cells with the protein kinase C(PKC) specific inhibitor-hypericin stimulation.Methods: Cultured human RPE cells were analyzed using the fluorescence Ca2+ dye fluo-3 AM and laser scanning confocal microscope(LSCM) after stimulation with 100nM phorbol 12-myristate 13-acetate(PMA) and (or)5 concentrations of hypericin(1, 2, 3, 4 and 5 μM).Results: The normal fluorescence in RPE cells was strong and distributed throughout the cells. The nucleus appeared to be more fluorescent than the cytoplasm. After stimulation with PMA alone or 5 concentrations of hypericin, a rapid decrease in flurescence intensity was observed. There was no obvious difference in decreased curve among 5concentrations. However, after stimulation with a 24 hr preincubation of PMA and 5 concentrations of hypericin, a further decrease was not observed.Conclusion: Fluo-3 AM appears to be a good indicator of the change in Ca2+ occurring in RPE cells and hypericin is a strong inhibitor of Ca2 + influx channel. Hypericin has potential as a therapeutic drug for proliferative vitreoretinopathy(PVR), the inhibitory effect on PVR might be caused by blocking the PKC activity and inhibiting Ca2+ influxpathway.

Hypericin in hypericum:chemistry, botanical sources and biological activities

Hypericin, a secondary metabolite first reported in 1830, is a natural photosensitizing naphtodianthrone and is mainly from the natural plant sources of genus Hypericum. Hypericin exhibits a wide variety of biological and pharmacological activities, such as antiviral, antidepressant, antitumor, antimicrobial, and antioxidant activity. Hypericin can also be utilized in photodynamic diagnosis. Accumulating evidence is pointing to the effects of hypericin with potential pharmaceutical and clinical interests in the past decades. The present review gives a comprehensive outline of the chemistry, botanical occurrence and biological activities of this powerful compound.

Raman spectroscopic study on structure of human immunodeficiency virus (HIV) and hypericin-induced photosen- sitive damage of HIV

The first Raman spectra of HIV1- HIV2 in human sera and hypericin-induced pho- tosensitive damage of the virus have been obtained. The prominent Raman lines in the spectra are assigned respectively to the carbohydrates of viral glycoprotein, RNA, protein and lipid. The spectra are dominated by Raman scattering of the carbohydrates. The lines of D-Mannose and N-acetylglucosamine in carbohydrates are obvious and there is a β-configuration in the anomeric C1 position in D-Mannose. The viral RNA duplexes bound assumes an A-form geometry. The lines of backbone phosphate group, bases (involving interbase hydrogen bonding) and ribose of the RNA are complete and distinct. The secondary structure of the viral protein maintains α-helix, β-sheet, β-turn and random coil. Its side chains are rich and vary from tryptophan, phenylalanine and “buried” tyrosine; the stable conformation of the S-S bond of gauche-gauche-gauche; the two forms of C-S bonds of gauche and trans ; to sulfhydrl group and ionized and unionized carboxyl groups. The viral lipid bilayer molecules are probably in the liquid ordered phase or the gel phase. It was observed that the hypericin-induced photosensitive damage of HIV1-HIV2 in human sera changed various components of HIV1-HIV2 in different degrees : The orderly A-form viral RNA would become a disordered viral RNA. There were a breakage of interbase hydrogen bonds and disruption of vertical base-base stacking interactions. In addition, the groups of ribos and four bases were damaged obviously. A decrease in ordered structure (α-helix and β-sheet) of viral protein is accompanied by an increase in random coil. The Tyr buried in the three-dimensional structure of protein was damaged, but it was still “buried” and the damage of C-S bond of trans form was stronger. The groups of carbohydrates, including D-Mannos and N-acetyl glucosamine, in viral envelope glycoprotein had also been changed. The hydrophilic C-N bond of choline in viral lipid was damaged, which was the possible binding site to hypericin, whereas the viral lipids bilayers were still probably in the liquid ordered phase or the gel phase. So the space structure of HIV1-HIV2 was damaged under the experimental conditions, which might block viral infection and inhibit its growth and breeding. It is apparent that the laser Raman spectra have provided certain direct evidence at the molecular level for photosensitive damage of HIV1-HIV2.

Signal interaction between nitric oxide and hydrogen peroxide in heat shock-induced hypericin production of Hypericum perforatum suspension cells

Heat shock(HS, 40℃, 10 min) induces hypericin production, nitric oxide(NO) generation, and hydrogen peroxide(H2O2) accumulation of Hypericum perforatum suspension cells.Catalase(CAT) and NO spe-cific scavenger 2-4-carboxyphenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide(cPTIO) suppress not only the HS-induced H2O2 generation and NO burst, but also the HS-triggered hypericin produc-tion.Hypericin contents of the cells treated with both NO and H2O2 are significantly higher than those of the cells treated with NO alone, although H2O2 per se has no effects on hypericin production of the cells, which suggests the synergistic action between H2O2 and NO on hypericin production.NO treatment enhances H2O2 levels of H.perforatum cells, while external application of H2O2 induces NO generation of cells.Thus, the results reveal a mutually amplifying action between H2O2 and NO in H.perforatum cells.CAT treatment inhibits both HS-induced H2O2 accumulation and NO generation, while cPTIO can also suppress H2O2 levels of the heat shocked cells.The results imply that H2O2 and NO may enhance each other's levels by their mutually amplifying action in the heat shocked cells.Membrane NAD(P)H oxidase inhibitor diphenylene iodonium(DPI) and nitric oxide synthase(NOS) inhibitor S,S′-1,3-phenylene-bis(1,2-ethanediyl)-bis-isothiourea(PBITU) not only inhibit the mutually amplifying action between H2O2 and NO but also abolish the synergistic effects of H2O2 and NO on hypericin production, showing that the synergism of H2O2 and NO on secondary metabolite biosynthesis might be dependent on their mutual amplification.Taken together, data of the present work demonstrate that both H2O2 and NO are essential for HS-induced hypericin production of H.perforatum suspension cells.Furthermore, the results reveal a special interaction between the two signal molecules in mediating HS-triggered secondary metabolite biosynthesis of the cells.

Hypericin: chemical synthesis and biosynthesis

Hypericin is one of the most important phenanthoperylene quinones extracted mainly from plants of the genus Hypericum belonging to the sections Euhypericum and Campylosporus of Keller's classification. Widespread attention to the antiviral and anti-tumor properties of hypericin has spurred investigations of the chemical synthesis and biosynthesis of this unique compound. However, the synthetic strategies are challenging for organic and biological chemists. In this review, specific significant advances in total synthesis, semi-synthesis, and biosynthesis in the past decades are summarized.

金丝桃素-β-环糊精络合物的制备及其在小龙虾保鲜膜中的应用

目的探究魔芋葡甘露聚糖(konjac glucomannan,KGM)、海藻酸钠(sodium alginate,SA)、巴西棕榈蜡(carnaubawax,CW)与金丝桃素-β-环糊精络合物(hypericin-β-cyclodextrin complex,Hyp-β-cd)制备的光敏性保鲜膜在4℃储藏条件下对小龙虾的品质影响。方法将制备的Hyp-β-cd加入到KGM、SA与CW溶液中,采用流延法制备光动力杀菌保鲜膜(KGM/SA/CW/Hyp-β-cd膜),通过核磁共振光谱(nuclear magnetic resonance spectroscopy,NMR)与荧光猝灭法验证了Hyp-β-cd的形成和活性氧(reativ oxygen species,ROS)的生成能力,然后用正交实验法对复合膜的制备进行优化。通过扫描电子显微镜(scanning electron microscope,SEM)、傅里叶红外光谱(Fourier transform infrared spectroscopy,FTIR)、X射线衍射(X-ray diffrction,XRD)进行表征,最后,以pH、总挥发性盐基氮(total volatile basic nitrogen,TVB-N)、菌落总数及硫代巴比妥酸反应物(thiobarbituric acid reactants,TBARS)值做为指标对保鲜效果进行评估。结果NMR及荧光猝灭的结果表明Hyp-β-cd的形成和ROS的生成,通过正交实验法对复合膜的制备进行优化:KGM含量为0.7%(m/V,下同)、SA含量为1.2%、CW含量为0.6%、Hyp-β-cd含量为0.4%时,复合膜有最优机械性能,拉伸强度为51.84MPa,断裂伸长率为14.27%。SEM证明了各物质在复合膜中的均匀分布。FTIR与XRD结果表明,随着SA、CW与Hyp-β-cd的加入,KGM与SA之间形成氢键,提高了复合膜的物理性能,CW的加入减少了复合膜的微孔隙,其中水接触角从67.5°增加到88.7°,氧气透过率从32.96降低到22.37 m^(3)·μm/m^(2)·day·kPa。由KGM/SA/CW/Hyp-β-cd膜包裹且在黄色LED光源照射处理后的小龙虾,在第8d时TVB-N值为20.37mg/g,菌落总数为6.14lgCFU/g,TBARS值为0.85 mg/kg,pH超过7.6。结论KGM/SA/CW/Hyp-β-cd复合膜具有作为光敏性抗菌包装材料的潜在应用价值。

金丝桃苷调节TXNIP/NLRP3信号通路对细菌性脑膜炎大鼠神经炎症的影响

目的:分析金丝桃苷(Hyp)调节硫氧还蛋白结合蛋白(TXNIP)/核苷酸结合寡聚结构域样受体蛋白3(NLRP3)信号通路对细菌性脑膜炎(BM)大鼠神经炎症的影响。方法:随机选择15只健康大鼠为健康组(尾静脉注射生理盐水),BM模型大鼠随机分为BM组(尾静脉注射生理盐水)、L-Hyp组(尾静脉注射10mg/kg Hyp)、H-Hyp组(尾静脉注射50mg/kg Hyp)、TXNIP-AAV组(尾静脉注射TXNIP-AAV)、AAV组(尾静脉注射AAV-NC)、白藜芦醇(Res)组(尾静脉注射30mg/kg Res)。采用Loeffler评分评估大鼠治疗后神经功能;血细胞分析仪测定白细胞(WBC)数量;酶联免疫吸附法分析脑脊液活性氧(ROS)、白细胞介素(IL)-1β、IL-6、肿瘤坏死因子α(TNF-α)水平;测定各组大鼠脑组织含水量;HE染色观察脑组织病理学变化;TUNEL染色观察脑组织细胞凋亡;免疫印迹法分析脑组织中TXNIP、NLRP3、caspase-1、凋亡相关斑点蛋白(ASC)、离子钙结合适配分子1(Iba1)、IL-1β蛋白表达。结果:与健康组比,BM组脑脊液ROS、WBC、IL-1β、IL-6、TNF-α水平及脑组织含水量、细胞凋亡率增加,Loeffler评分减少(P<0.05);与BM组比,L-Hyp组、H-Hyp组Loeffler评分增加,脑脊液ROS、WBC、IL-1β、IL-6、TNF-α水平及脑组织含水量、细胞凋亡率减少(P<0.05);与H-Hyp组比,TXNIP-AAV组、AAV组脑脊液ROS、WBC、IL-1β、IL-6、TNF-α水平及脑组织含水量、细胞凋亡率增加,Loeffler评分减少,Res组Loeffler评分增加,脑脊液ROS、WBC、IL-1β、IL-6、TNF-α水平及脑组织含水量、细胞凋亡率减少(P<0.05)。健康组大鼠脑组织形态正常;BM组、TXNIP-AAV组、AAV组脑组织形态改变,细胞排列散乱,出现核皱缩;L-Hyp组、H-Hyp组、Res组脑组织形态有所改善,细胞坏死、皱缩减少。与健康组比,BM组脑组织中TXNIP、NLRP3、caspase-1、ASC、Iba1、IL-1β表达增加(P<0.05);与BM组比,LHyp组、H-Hyp组脑组织中TXNIP、NLRP3、caspase-1、ASC、Iba1、IL-1β表达减少(P<0.05);与H-Hyp组比,TXNIP-AAV组、AAV组脑组织中TXNIP、NLRP3、caspase-1、ASC、Iba1、IL-1β表达增加,Res组脑组织中TXNIP、NLRP3、caspase-1、ASC、Iba1、IL-1β表达减少(P<0.05)。结论:Hyp可能抑制BM大鼠神经炎症,其机制可能与抑制TXNIP/NLRP3通路有关。

中医情志病抑郁障碍的发病机制及贯叶连翘在抗抑郁领域中的研究进展

随着社会高速发展,抑郁症的患病率、致残率、致死率日益增高。其发病机制具有复杂性,异质性,尚未完全了解。目前的治疗方法机制不同效,正在广泛探讨中。本文总结了中医情志病抑郁障碍的发病机理和中医药抗抑郁的研究进展,着重阐述了贯叶连翘及其提取物金丝桃素的抗抑郁效果和分子机制。

兴安杜鹃3种主要成分的HPLC测定及其对大鼠气管平滑肌的舒张作用

[目的]建立同时测定兴安杜鹃中金丝桃苷、槲皮素和杜鹃素含量的高效液相色谱法(HPLC),并探讨兴安杜鹃提取物及3种主要成分对大鼠气管平滑肌舒张作用的影响及其作用机制。[方法]采用HPLC法测定兴安杜鹃中金丝桃苷、槲皮素和杜鹃素的含量并进行方法学考察。采用离体气管灌流法取大鼠气管环,将气管环固定于麦氏浴皿中,使用扑尔敏、吲哚美辛、格列苯脲、左旋硝基精氨酸甲酯(L-NAME)、硝苯地平和普萘洛尔进行预孵育,待气管环张力稳定后,加入60 mmol/L KCl溶液收缩气管环,进行兴安杜鹃对气管环的舒张试验,观察大鼠气管平滑肌张力变化。[结果]兴安杜鹃中3种主要成分HPLC测定方法考察结果显示,3种成分均在2~80μg/mL线性关系良好,平均回收率分别为金丝桃苷100.3%(RSD=2.63%)、槲皮素102.7%(RSD=2.89%)、杜鹃素102.1%(RSD=2.36%)。在吲哚美辛、格列本脲和普萘洛尔孵育组织中,兴安杜鹃提取物及3种单体成分的舒张作用与未孵育组相比没有显著差异;在扑尔敏、L-NAME和硝苯地平孵育的组织中,兴安杜鹃及金丝桃苷与未孵育的组织相比有显著差异,舒张作用有所降低。这种现象可能是通过抑制组胺H1受体、一氧化氮合酶途径和钙离子通道介导的钙离子内流实现的。[结论]该试验建立的方法简单、准确,能快速测定兴安杜鹃中金丝桃苷、槲皮素和杜鹃素的含量,为其质量评价和控制提供了有效的分析手段;并且这3种成分与兴安杜鹃舒张气管的作用有关,可作为支气管扩张剂开发的理论依据。

金丝桃素通过调控AMPK/Nrf2/HO-1信号通路减轻大鼠心肌缺血再灌注损伤

目的:探讨金丝桃素(hypericin,Hyp)对心肌缺血再灌注损伤(myocardial ischemia-reperfusion injury,MIRI)大鼠心脏的影响及其作用机制。方法:30只SPF级雄性SD大鼠随机分为5组:假手术组(sham组)、心肌缺血再灌注损伤组(MIRI组)、金丝桃素低剂量给药组(MIRI+L-Hyp组)、金丝桃素高剂量给药组(MIRI+H-Hyp组)以及阳性对照药物曲美他嗪(trimetazidine,TMZ)给药组(MIRI+TMZ组),每组6只。除假手术组,其余4组大鼠均采用结扎左冠状动脉前降支后再通法建立MIRI模型,通过监测心电图判断造模是否成功;利用超声心动图检测大鼠心功能;TTC染色检测大鼠心肌梗死面积;HE染色观察大鼠心肌形态学特征;应用生化试剂盒分别测定大鼠血清中乳酸脱氢酶(lactate dehydrogenase,LDH)、超氧化物歧化酶(superoxide dismutase,SOD)的活性以及丙二醛(malondialdehyde,MDA)的含量;应用ELISA试剂盒分别检测大鼠血清中心肌肌钙蛋白(cardiac troponin I,cTnI)和活性氧(reactive oxygen species,ROS)含量;Western blot测定大鼠心肌组织中腺苷酸激活蛋白激酶(AMP-activated protein kinase,AMPK)、p-AMPK、核因子E2相关因子2(nuclear factor E2-related factor 2,Nrf2)以及血红素加氧酶1(heme oxygenase-1,HO-1)蛋白表达水平。结果:与sham组相比,MIRI组大鼠心肌组织病理损伤加剧,心肌梗死面积增大,超声心动图显示左心室射血分数(left ventricular ejection fraction,LVEF)和左心室短轴缩短率(left ventricular fractional shortening,LVFS)降低,LDH活性、cTnI、MDA以及ROS含量均升高,SOD活性、p-AMPK、Nrf2、HO-1蛋白表达均显著降低(P<0.05);与MIRI组相比,金丝桃素低、高剂量给药组及阳性对照药物曲美他嗪组大鼠心肌组织病理损伤减轻及心肌组织梗死面积减少,LVEF及LVFS升高,血清中LDH活性、cTnI、MDA以及ROS含量降低,而SOD活性、p-AMPK、Nrf2、HO-1蛋白表达均升高(P<0.05)。结论:金丝桃素可减轻大鼠心肌缺血再灌注损伤,可能通过调节AMPK/Nrf2/HO-1信号通路发挥作用。

Ultrastructure of the Multicellular Nodules in Hypericum perforatum Leaves

With the development of the multicellular nodules in the leaves of Hypericum per plastids increased both in number and volume, and some plastids degenerated and were surrounded by dark tubular elements and vesicles. Some vesicles fused with vacuoles and secreted substances into vacuoles. There are many multivesicles, multimembrane structures and osmiophilic droplets in vacuoles of nodule cells. Meanwhile, dictyosome secreted vesicles into the vacuoles. However, there is a large central vacuole completely filled with secretory materials (hypericin) in matured multicellular nodules. This suggests that the osmiophilic droplets may be a precursor of hypericin originated from the degenerated plastids. There were abundant endoplasmic reticulums and dictyosomes between the plastids and osmiophilic droplets, suggesting that they may be involved in the synthesis and/or transport of the precursor of hypericin.

金丝桃素与乙基金丝桃素的全合成及对人免疫缺陷病毒逆转录酶的抑制活性

金丝桃素与乙基金丝桃素的全合成及对人免疫缺陷病毒逆转录酶的抑制活性赵晶张致平陈鸿珊陈湘红（中国医学科学院医药生物技术研究所，北京１０００５０）近年发现由贯叶金丝桃（Ｈｙｐｅｒｉｃｕｍｐｅｒｚｏｒａｔｕｍ），三棱三叶金丝桃（Ｈ．ｔｒｉｑｕｅｔｒｉｆ...

金丝桃属植物研究进展

金丝桃属植物中许多种是传统草药,二蒽酮类、类黄酮类及间苯三酚类等次生代谢产物具有许多药理作用.因此近年来对该属植物的分泌结构、具药理作用的化学成分、快速繁殖以及分子水平的研究越来越多.本文就以上各方面的研究进展作一综述.