墨江蜈蚣与少棘蜈蚣的比较研究：I.化学组成

本文对墨江蜈蚣（Scolopendra mojiangica Zhang et Chi）与少棘蜈蚣（Scolopendra subspinipes mutilans L.Koch）的水分、灼烧残渣、蛋白质、游离氨基酸、微量元素和挥发性脂肪酸的含量进行了系统的测定并比较了两者间的异同性。实验结果表明：墨江蜈蚣的蛋白质、灼烧残渣、水分的含量分别为67.2%、3.95%和3.59%,而少棘蜈蚣分别为68.8%、4.80%和3.65%,它们的含量基本相同。游离氨基酸、挥发性脂肪酸和微量元素的相对含量也很接近,但脂类和总糖的含量差异较大。     

墨江蜈蚣与少棘蜈蚣的比较研究──Ⅰ．化学组成

本文对墨江蜈蚣（ＳｃｏｌｏｐｅｎｄｒａｍｏｊａｎｇｉｃａＺｈａｎｇｅｔＣｈｉ）与少棘蜈蚣（ＳｃｏｔｏｐｅｎｄｒａｓｕｂｓｐｉｎｉｐｅｓｍｕｔｉｌａｎｓＬ．Ｋｏｃｈ）的水分、灼烧残渣、蛋白质、游离氨基酸、微量元素和挥发性脂肪酸的含量进行了系统的测定并比较了两者间的异同性。实验结果表明：墨江蜈蚣的蛋白质、灼烧残渣、水分的含量分别为６７．２％、３．９５％和３．５９％，而少棘蜈蚣分别为６８．８％、４．８０％和３．６５％，它们的含量基本相同。游离氨基酸、挥发性脂肪酸和微量元素的相对含量也很接近，但脂类和总糖的含量差异较大。     

墨江蜈蚣与少棘蜈蚣的比较研究：II.药效和毒理

本文比较了墨江蜈蚣与少棘蜈蚣的药效学和毒理学作用,实验包括抗惊厥试验、对致病性真菌和细菌体外生长的影响、急性毒性试验和染色体畸变实验等。结果两种蜈蚣的３％醋酸提取液对所试的真菌有抑制作用但无杀真菌作用,而对细菌无抑制作用。两种蜈蚣的毒性很低,在给小鼠５０ｇ／ｋｇ的剂量下都无法测出ＬＤ５０。在２０５ｍｇ／ｋｇ条件下的致突变率与未给药的突变率接近,表明两者都无遗传毒性。     

少棘巨蜈蚣的化学组成

在我国蜈蚣作为药用已有数百年历史,在《本草纲目》等古代及现代医药书籍中多有描述。     

少棘蜈蚣候选杀虫肽κ-SLPTX-Ssm2e的原核表达与纯化

κ-SLPTX-Ssm2e是一条通过生物信息学同源序列分析获得的与已知具有杀虫活性的毒素κ-SLPTX-Ssm2a高度同源的少棘蜈蚣毒素,由31个氨基酸残基组成,且含有3对二硫键。为了后续便于对其结构和功能进行研究,现采用原核表达与纯化的方法来获取较高产量的毒素多肽。文中首先利用大肠杆菌自动诱导表达系统,在大肠杆菌表达菌株BL21(DE3)中表达融合蛋白GST-SUMO-κ-SLPTX-Ssm2e,将其通过GST亲和层析纯化后,采用Ulp1激酶酶切,以释放毒素分子。随后,通过反相高效液相色谱(RP-HPLC)进一步纯化。最后,利用MALDI-TOF/TOF质谱仪鉴定目的多肽峰的相对分子质量为3 475.732,与κ-SLPTX-Ssm2e的理论相对分子质量3 475.07基本一致,说明κ-SLPTX-Ssm2e很有可能被成功表达。上述结果为进一步研究κ-SLPTX-Ssm2e的杀虫活性奠定了基础。     

两种药用蜈蚣的简介

两种药用蜈蚣的简介张崇洲，王克勤（中国科学院动物研究所北京１０００８０）（湖北省中医药研究院中药研究所）关键词少棘蜈蚣，多棘蜈蚣，区别蜈蚣，迄今常用的多足动物名称，在最早记载它的《广雅》里写为“吴公”。蜈蚣作为一味动物药在我国最古老的药典《神农本草经...     

中国少棘蜈蚣（Scolopendra subspinipes mutillans）的毒腺结构

用免疫组化和光镜、透射电镜等观察了中国少棘蜈蚣毒腺的结构。结果显示,纵贯颚肢的弯月形毒腺为单管泡状腺,主要由柱状分泌细胞和介于其间的纤细表皮细胞组成。被肌肉束环绕的分泌细胞辐射状排列于几丁质的毒液导管外,其纤细的颈部由环状括约肌控制,分泌端以折叠回转的单向瓣膜经导管壁上的孔道直接伸入管腔,膨大的盲端直达毒腺底膜。高电子密度的分泌溶酶体向分泌口汇集时电子密度逐渐降低并降解为分泌小泡,其中的杆状结晶样毒蛋白也经无定型状态逐渐分散,经胞吐作用进入管腔并进一步疏散和均质化。免疫组化显示,分泌细胞颈部密集的分泌颗粒的主要成分为毒蛋白,毒蛋白在分泌细胞中呈明显的向心式梯度增强型分布。根据上述观察,提出了蜈蚣毒液以分泌溶酶体介导的非经典途径分泌的观点。     

火棘多糖的分离纯化及抗氧化活性研究

对火棘多糖的提取工艺、分离纯化以及抗氧化活性作了研究.结果表明:火棘多糖的最佳提取工艺为90℃、9h,料液比为1∶10,提取率3.5%;火棘多糖具有明显的体外清除含氧自由基的作用,DPPH·分光光度法测得SC50为1.87mg/mL;通过分级沉淀、DE-52柱层析分离得到PP-A2、PP-A3、PP-A4和PP-B1、PP-B2、PP-B36种级分;经SephadexG-200柱层析鉴定,PP-A2、PP-A3、PP-B2为均一多糖.     

蜈蚣碱性蛋白SSmp-d的分离纯化及其部分理化性质的鉴定

少棘巨蜈蚣（ＳｃｏｌｏｐｅｎｄｒａｓｕｂｓｐｉｎｉｐｅｓｍｕｔｉｌａｎｓＬ．Ｋｏｃｈ）经９５％乙醇脱脂后,再经４℃水冷渗,水提液低温旋转浓缩,冻干,得到的冻干粉先后经过ＳｅｐｈａｄｅｘＧ－２５柱,等电聚焦制备电泳,再经ＳｅｐｈａｄｅｘＧ－１５０柱,ＳｅｐｈａｄｅｘＧ－１００柱,最后经ＨＰＬＣ制备得到一个纯的碱性蛋白,命名为ＳＳｍｐ－ｄ．该蛋白经ＨＰＬＣ、超薄等电聚焦电泳检验是均一的．采用ＨＰＬＣ和Ｐｒｏｔｅｉｎ－ＰａｋＴＭ１２５柱测定其分子量为２４．６４ｋＤ．ＩＥＦ－ＨＰＣＥ显示其等电点为９．２７．氨基酸分析表明ＳＳｍｐ－ｄ含较多的Ａｒｇ、Ｌｙｓ等碱性氨基酸,另外还含有较多的Ａｌａ、Ｌｅｕ．使用蛋白质自动序列分析仪测定了ＳＳｍｐ－ｄＮ端的１１个氨基酸,序列为ＮＨ３＋－Ａｓｐ－Ｖａｌ－Ａｓｎ－Ｐｈｅ－Ａｒｇ－Ｌｅｕ－Ｓｅｒ－Ｇｌｙ－Ａｌａ－Ａｓｐ－Ｐｒｏ．     

蜈蚣碱性蛋白SSmp—d的分离纯化及其部分理化性质的鉴定

少棘巨蜈蚣经９５％乙醇脱脂后,再经４℃水冷渗,水提液低温旋转浓缩,冻干,得到的冻干粉先后经过Ｓｅｐｈａｄｅｘ Ｇ－２５柱,等电聚焦制备电泳,再经Ｓｅｐｈａｄｅｘ Ｇ－１５０柱,Ｓｅｐｈａｄｅｘ Ｇ－１００柱,最后经ＨＰＬＣ制备得到一个纯的碱性蛋白,命名为ＳＳｍｐ－ｄ。该蛋白经ＨＰＬＣ、超薄等电聚焦电泳检验是均一的。采用ＨＰＬＣ和Ｐｒｏｔｅｉｎ－Ｐａｋ＾ＴＭ１２５柱测定其分子量为２４．６４ｋＤ。ＩＥ     

Centipede (Scolopendra subspinipes mutilans) venom toxin peptide exhibits cytotoxic and cell growth effects in a concentration-dependent manner

Centipede venom contains a variety of proteins, peptides, and enzymes. However, the biological actions of toxin peptides in centipede venom remain largely unknown. In this study, we identified a centipede (Scolopendra subspinipes mutilans) venom toxin peptide (SsmTP) that was shown to act as a cell growth factor at low concentrations-in vitro. SsmTP was found to consist of 66 amino acids that display seven cysteine residues, which exhibited high similarity to the predicted neurotoxin. SsmTP was expressed in the venom gland of S. s. mutilans. A recombinant SsmTP peptide of approximately 5.2 kDa was produced in baculovirus-infected insect cells. Interestingly, SsmTP exhibited cytotoxicity in murine cells in a concentration-dependent manner but displayed a cell growth effect at low concentrations. SsmTP at a low concentration also protected murine cells against oxidative damage through the inhibition of caspase-1 and apoptosis. Our data indicate that SsmTP acts as a cell growth factor and a toxin peptide in a concentration-dependent manner. Consequently, our results provide evidence that the identification of SsmTP, a centipede toxin peptide, may not only elucidate the biological action of toxin peptides but also offer additional insight into the pharmacological applications of centipede venoms.     

A New 1,5‐Dihydroxy‐4‐methoxyisoquinoline from Scolopendra subspinipes mutilans

A new isoquinoline, 1,5‐dihydroxy‐4‐methoxyisoquinoline ( 1 ), was obtained from Scolopendra subspinipes mutilans. Compound 1 showed moderate cytotoxicity on tumour cells with IC₅₀ values ranging from 13 to 26 μm against five esophageal squamous cancer cells whereas low cytotoxicity against normal human esophageal epithelial cells. Isoquinoline ring oxidized at C(1), C(4), and C(5) can enhance its cytotoxicity. In addition, compound 1 showed potent inhibitory effect (inhibition rate > 50% at 13 μm ) on cell migration in human umbilical vein endothelial cells. This article mainly studies the structure and activity of 1 , and more modification of 1 as a potential anticancer agent.     

Centipede venom peptide SsmTX‐I with two intramolecular disulfide bonds shows analgesic activities in animal models

Pain is a major symptom of many diseases and results in enormous pressures on human body or society. Currently, clinically used analgesic drugs, including opioids and nonsteroidal anti‐inflammatory drugs, have adverse reactions, and thus, the development of new types of analgesic drug candidates is urgently needed. Animal venom peptides have proven to have potential as new types of analgesic medicine. In this research, we describe the isolation and characterization of an analgesic peptide from the crude venom of centipede, Scolopendra subspinipes mutilans. The amino acid sequence of this peptide was identical with SsmTX‐I that was previously reported as a specific Kv2.1 ion channel blocker. Our results revealed that SsmTX‐I was produced by posttranslational processing of a 73‐residue prepropeptide. The intramolecular disulfide bridge motifs of SsmTX‐I was Cys1–Cys3 and Cys2–Cys4. Functional assay revealed that SsmTX‐I showed potential analgesic activities in formalin‐induced paw licking, thermal pain, and acetic acid‐induced abdominal writhing mice models. Our research provides the first report of cDNA sequences, disulfide motif, successful synthesis, and analgesic potential of SsmTX‐I for the development of pain‐killing drugs. It indicates that centipede peptide toxins could be a treasure trove for the search of novel analgesic drug candidates. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.     

Antimicrobial peptide scolopendrasin VII,derived from the centipede Scolopendra subspinipes mutilans,stimulates macrophage chemotaxis via formyl peptide receptor 1

In this study, we report that one of the antimicrobial peptides scolopendrasin VII, derived from Scolopendra subspinipes mutilans, stimulates actin polymerization and the subsequent chemotactic migration of macrophages through the activation of ERK and protein kinase B (Akt) activity. The scolopendrasin VII-induced chemotactic migration of macrophages is inhibited by the formyl peptide receptor 1 (FPR1) antagonist cyclosporine H. We also found that scolopendrasin VII stimulate the chemotactic migration of FPR1-transfected RBL-2H3 cells, but not that of vector-transfected cells; moreover, scolopendrasin VII directly binds to FPR1. Our findings therefore suggest that the antimicrobial peptide scolopendrasin VII, derived from Scolopendra subspinipes mutilans, stimulates macrophages, resulting in chemotactic migration via FPR1 signaling, and the peptide can be useful in the study of FPR1-related biological responses. [BMB Reports 2015; 48(8): 479-484]     

Induction, purification and characterization of an antibacterial peptide scolopendrin I from the venom of centipede Scolopendra subspinipes mutilans

The crude venom of the centipede Scolopendra subspinipes mutilans, injected with Escherichia coli K12D31 for 3-4 days showed broad-spectrum antimicrobial activity against Gram-positive. Gram-negative bacteria and fungi. It showed good antibacterial activity against E. coli K12D31 at different temperatures, pH, and ionic strengths. The crude venom was heated at 100 degrees C for 30 min, centrifuged at 10,000 rpm for 30 min at 4 degrees C and the supernatants were obtained, from which an antibacterial fraction having a molecular mass of 3000-5000 Da, was further separated by ultrafiltration. A homogeneous antibacterial peptide named scolopendrin I, having a molecular mass of 4,498 Da, was isolated using cation-exchange chromatography and two steps of reverse-phase high performance liquid chromatography (RP-HPLC). Scolopendrin I did not show any hemolytic and agglutination activities at the concentration below 30 microM.     

Purification and molecular cloning of a novel serine protease from the centipede, Scolopendra subspinipes mutilans

A novel serine protease, named as scolonase, was purified and characterized from the tissue of the Korean centipede, Scolopendra subspinipes mutilans. Purified scolonase showed an apparent molecular weight of 25 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and an isoelectric point of 4.8 on isoelectric focusing gel. Scolonase was able to preferentially hydrolyze arginine over lysine at the cleavage site among the several synthetic peptide substrates. Scolonase has also a potent fibrinolytic activity by converting human Glu-plasminogen to activated plasmin due to the specific cleavage of the molecule at the peptide bond Arg(561)-Val(562). The enzyme activity of scolonase was completely inhibited by phenylmethanesulfonyl fluoride and difluorophosphate. The cDNA encoding scolonase was cloned from the cDNA library of the centipede constructed with oligonucleotide probe, which was designed on the basis of the N-terminal amino acid sequence of scolonase. The deduced complete amino acid sequence of scolonase demonstrated that the protein is composed of 277 amino acids including 33 amino acids as a leader sequence, and that it has significant sequence homology with other serine proteases.     

Prey orientation and the role of venom availability in the predatory behaviour of the centipede Scolopendra subspinipes mutilans (Arthropoda: Chilopoda)

Many animal phyla contain clades in which most or all species are venom-injecting predators. An example, in the arthropods, is the class Chilopoda, containing the approximately 3500 species of centipedes. Very little ecological or behavioural work yielding quantitative data has been conducted on centipede predation. Here, we describe a study of this kind. Our experiments employed one centipede species - a large tropical one, Scolopendra subspinipes mutilans - and two species of prey - a cricket, Gryllus assimilis, and a locust, Schistocerca gregaria. We conducted two experiments. The first was aimed at investigating the extent to which the centipedes attacked prey in particular tagmata as opposed to at random over the whole body surface. The results showed that the centipedes were highly selective, preferring to attack the head or thorax rather than the abdomen; indeed, they often reoriented the prey in order to achieve this. A possible explanation of this behaviour is to maximize the speed with which the neurotoxins in the venom reach either the brain or the thoracic ganglia that control limb movement. The second experiment was aimed at investigating the effect of venom-extraction on the attack rate, and specifically at testing if the magnitude of any such effect differed between the two types of prey, which differ considerably in size. The results showed a major effect of venom extraction in relation to both types of prey, but with the time taken to return to a 'normal' attack rate being longer in the case of the larger prey-type, namely the locust. We discuss these results in relation to the 'venom optimization hypothesis' and, more generally, to the principle of minimizing the production/use of venom, which is an energetically expensive resource.     

Identification and optimization of an antimicrobial peptide from the ant venom toxin pilosulin

A template based on positional residue frequencies in the N-terminal stretch of natural alpha-helical antimicrobial peptides was used to prepare sequence patterns and to scan the Swiss-Prot Database, using the ScanProsite tool. This search identified a segment in pilosulin 1, a cytotoxic peptide from the venom of the jumper ant Myrmecia pilosula, as a potential novel antimicrobial peptide sequence. This segment, corresponding to the 20 N-terminal residues, was synthesized and its structural properties and biological activities were investigated. It showed a potent and broad spectrum antimicrobial activity including standard and multi-drug resistant gram-positive and gram-negative bacteria and Candida albicans, confirming the validity of the search method. A rational redesign approach resulting in four amino acid substitutions yielded a variant with improved antibacterial and significantly reduced hemolytic activity.     

蜈蚣(Scolopendra subspinipes mutilans L.Koch)毒的化学组成和生物活性

系统分析少棘蜈蚣粗毒的化学组成,蛋白质含量86.23％、水不溶物质0.24％、还原糖0.23％、水份2.1％,仅含Ser、Pro、Arg等三种游离氨基酸,鉴定了11种微量元素,碱性凝胶电泳显示20条蛋白质谱带。分析了10种酶的活性、出血毒性及对血小板聚集的影响,其中精氨酸酯酶活力最高,不存在类凝血酶、淀粉酶活性及出血毒性,蜈蚣毒的浓度为0.3μg/μL时强烈诱导血小板的聚集。