Identification and characterization of hirudin-HN, a new thrombin inhibitor, from the salivary glands of<i>Hirudo nipponia</i>

Cheng, Biao; Liu, Fei; Q, Guo; Lu, Yinghua; Shi, Hong-Zhuan; Ding, Andong; Xu, Chengfeng

doi:10.7717/peerj.7716

Cited by 20 publications

(27 citation statements)

References 66 publications

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“…Hirudin deactivates thrombin by simultaneously binding to the active site to form irreversible stoichiometric complexes. 31 qRT-PCR confirmed that the level of miR-24 increased after ICH in ECs, which were isolated by LCM. EC proliferation was also observed.…”

Section: Thrombin Elevated Mir-24 Expression and Angiogenesis After Ichmentioning

confidence: 67%

Thrombin-induced miRNA-24–1-5p upregulation promotes angiogenesis by targeting prolyl hydroxylase domain 1 in intracerebral hemorrhagic rats

Cui

Yang

Zhou

et al. 2021

Journal of Neurosurgery

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OBJECTIVEThrombin is a unique factor that triggers post-intracerebral hemorrhage (ICH) angiogenesis by increasing hypoxia-inducible factor–1α (HIF-1α) at the protein level. However, HIF-1α mRNA remains unchanged. MicroRNAs (miRNAs) mediate posttranscriptional regulation by suppressing protein translation from mRNAs. This study aimed to determine if miRNAs might be involved in thrombin-induced angiogenesis after ICH by targeting HIF-1α or its upstream prolyl hydroxylase domains (PHDs).METHODSThe study was divided into two parts. In part 1, rats received an injection of thrombin into the right globus pallidus. An miRNA array combined with miRNA target prediction, luciferase activity assay, and miRNA mimic/inhibitor transfection were used to identify candidate miRNAs and target genes. Part 2 included experiments 1 and 2. In experiment 1, rats were randomly divided into the sham group, ICH group, and ICH+hirudin–treated (thrombin inhibitor) group. In experiment 2, the rats were randomly divided into the sham group, ICH group, ICH+antagomir group, ICH+antagomir-control group, and ICH+vehicle group. Western blotting and quantitative real-time polymerase chain reaction were used to determine the expression of protein and miRNA, respectively. The coexpression of miR-24–1-5p (abbreviated to miR-24) and von Willebrand factor was detected by in situ hybridization and immunohistochemical analysis. The angiogenesis was evaluated by double-labeling immunofluorescence. Neurological function was evaluated by body weight, modified Neurological Severity Scores, and corner turn and foot-fault tests.RESULTSIn part 1, it was shown that miR-24, which is predicted to target PHD1, was upregulated (fold-change of 1.83) after thrombin infusion, and that the miR-24 mimic transfection decreased luciferase activity and downregulated PHD1 expression (p < 0.05). miR-24 inhibitor transfection increased PHD1 expression (p < 0.05). In part 2, it was shown that miR-24 was expressed in endothelial cells. The HIF-1α protein level and proliferating cell nuclear antigen–positive (PCNA+) nuclei in vessels were increased, while the PHD1 protein level was decreased after ICH, and these effects were reversed by hirudin (p < 0.05). The antagomiR-24–treated rats exhibited a markedly lower body weight and significantly poorer recovery from neurological deficit compared with those in ICH groups (p < 0.05). AntagomiR-24 intervention also led to lower miR-24 expression, a higher PHD1 protein level, and fewer PCNA+ nuclei in vessels compared with those in ICH groups (p < 0.05).CONCLUSIONSThe present study suggests that thrombin reduces HIF-1α degradation and initiates angiogenesis by increasing miR-24, which targets PHD1 after ICH.

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Section: Thrombin Elevated Mir-24 Expression and Angiogenesis After Ichmentioning

confidence: 67%

Thrombin-induced miRNA-24–1-5p upregulation promotes angiogenesis by targeting prolyl hydroxylase domain 1 in intracerebral hemorrhagic rats

Cui

Yang

Zhou

et al. 2021

Journal of Neurosurgery

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“…Therefore, coagulation is retarded or completely prevented relying on the hirudin concentration in blood. 11 , 12 In Figure 1 previous research of LSE demonstrated that it contains a wide range of active proteins with diverse molecular weight.…”

Section: Introductionmentioning

confidence: 97%

Time to Change Theory; Medical Leech from a Molecular Medicine Perspective Leech Salivary Proteins Playing a Potential Role in Medicine

Shakouri

Wollina

2020

Adv Pharm Bull

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Followed by developing modern medicine, leeches did not have extensive use as before; however, in the late 19th century, they were still used in most countries all over the world. Thus far, leeches were utilized in treating various diseases like skin disorders, arthritis, and cancer. In Egypt, using leeches for treatment dates back to early 1500 BC. A medical leech’s salivary glands involve over 100 bioactive proteins and the salivary gland secretion contains bacteriostatic, analgesic, and anticoagulation influences; with resolving activity, it causes microcirculation disorders elimination, restoring the hurt vascular permeability of organs and tissues, removing hypoxia, decreasing blood pressure and detoxifying the organism by antioxidant paths. The current work reviews the innovative treatment with medical leech, especially proteins in leech saliva extraction (LSE) with high potential in medicine. The virtue of salivary gland secretions which are proteinaceous enzymes, leech acts on various diseases such as venous congestion in reconstructive and plastic surgery, osteoarthritis, cardiovascular diseases caused by blood coagulation disorders, pain management, priapism, macroglossia, cancer complications, wounds and many other. To confirm the potential therapeutic impacts of leech treatment, more studies are required in more extensive areas with more exact methodologies.

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“…Exploring the diversity of anticoagulant-related proteins and framing these in an evolutionary context in terms of presence or absence of orthologues across the phylogeny of leeches would help evince the general evolution of these proteins. Members of several leech families have been investigated at the transcriptome level for compositions of the anticoagulant repertoires, including members of Piscicolidae, Glossiphoniidae, Macrobdellidae, Haemadipsidae, Praobdellidae and Hirudinidae 3,8,[15][16][17][18][19][20][21][22][23][24] . Genome-level evaluations within a species like H. medicinalis would allow insights into the diversity of bioactive proteins available.…”

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confidence: 99%

Draft genome of the European medicinal leech Hirudo medicinalis (Annelida, Clitellata, Hirudiniformes) with emphasis on anticoagulants

Kvist

Manzano-Marı́n

Carle

et al. 2020

Sci Rep

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the european medicinal leech has been used for medicinal purposes for millennia, and continues to be used today in modern hospital settings. its utility is granted by the extremely potent anticoagulation factors that the leech secretes into the incision wound during feeding and, although a handful of studies have targeted certain anticoagulants, the full range of anticoagulation factors expressed by this species remains unknown. Here, we present the first draft genome of the European medicinal leech, Hirudo medicinalis, and estimate that we have sequenced between 79-94% of the full genome. Leveraging these data, we searched for anticoagulation factors across the genome of H. medicinalis. following orthology determination through a series of BLASt searches, as well as phylogenetic analyses, we estimate that fully 15 different known anticoagulation factors are utilized by the species, and that 17 other proteins that have been linked to antihemostasis are also present in the genome. We underscore the utility of the draft genome for comparative studies of leeches and discuss our results in an evolutionary context. Notwithstanding the significance of the European medicinal leech Hirudo medicinalis Linnaeus, 1758 as a medical tool from antiquity through modern science-based medicine, it is remarkable that this species' repertoire of anticoagulants and other bioactive salivary proteins has escaped published in-depth investigation (but see 1). In order to maintain blood flow in the tissues surrounding the incision wounds during their extended periods of feeding and, importantly, keeping the blood from clotting inside of the crop during the periods of digestion, bloodfeeding leeches secrete a variety of bioactive compounds from their salivary glands. Over 20 different compounds relating to antihemsotasis have been isolated from leech saliva including, for example, direct thrombin inhibitors, factor Xa inhibitors, trypsin inhibitors, endoglucuronidases and antiplatelet proteins 2,3. Despite its historical significance in medicine, the detailed composition of the anticoagulant profile of Hirudo medicinalis Linnaeus, 1758 has escaped scrutiny insofar as modern studies have been concerned primarily with the commercially available species Hirudo verbana (Carena, 1820) 4-7 ; but see also 1,8. Whereas H. medicinalis was long thought to be the centerpiece for the clinical application of leeches in hospital settings, recent investigations have shown that Hirudo verbana is far more commonly employed for this purpose 4 yet sold under the name H. medicinalis; this confusion also prompted the designation of a neotype specimen for H. medicinalis in order to ameliorate future taxonomic confusion 9. The utility of leeches in modern medicine focuses on post-surgical procedures, chiefly following digit replantation or skin grafting surgery 10,11 wherein the build-up of blood in the extremities and resulting congestion of the veins is relieved by their application. By contrast, medieval medicinal practices,

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Identification and characterization of hirudin-HN, a new thrombin inhibitor, from the salivary glands ofHirudo nipponia

Cited by 20 publications

References 66 publications

Thrombin-induced miRNA-24–1-5p upregulation promotes angiogenesis by targeting prolyl hydroxylase domain 1 in intracerebral hemorrhagic rats

Thrombin-induced miRNA-24–1-5p upregulation promotes angiogenesis by targeting prolyl hydroxylase domain 1 in intracerebral hemorrhagic rats

Time to Change Theory; Medical Leech from a Molecular Medicine Perspective Leech Salivary Proteins Playing a Potential Role in Medicine

Draft genome of the European medicinal leech Hirudo medicinalis (Annelida, Clitellata, Hirudiniformes) with emphasis on anticoagulants

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