Purification and characterization of a novel anti-coagulant from the leech Hirudinaria manillensis

Cheng, Ruomei; Tang, Xiaopeng; Long, Ai-Lin; Mwangi, James; Lai, Ren; Sun, Rui-Pu; Long, Chengbo; Zhang, Zhenqing

doi:10.24272/j.issn.2095-8137.2019.037

Cited by 11 publications

(7 citation statements)

References 30 publications

(43 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Several studies have established that the anti-thrombotic effects of leeches are associated with the bioactive polypeptides and proteins. The biological activities of these proteins and polypeptides can vary because of differences in the amino acid composition and molecular weights ( Tang et al, 2018 ; Cheng et al, 2019 ). In this study, we prepared APP, APH, and APA using bio-mimetic enzymatic hydrolysis.…”

Section: Discussionmentioning

confidence: 99%

Characterization of active peptides derived from three leeches and comparison of their anti-thrombotic mechanisms using the tail vein thrombosis model in mice and metabonomics

Dong,

Li,

et al. 2024

Front. Pharmacol.

View full text Add to dashboard Cite

Background and aims: The increasing incidence of cardiovascular diseases has created an urgent need for safe and effective anti-thrombotic agents. Leech, as a traditional Chinese medicine, has the effect of promoting blood circulation and removing blood stasis, but its real material basis and mechanism of action for the treatment of diseases such as blood stasis and thrombosis have not been reported.Methods: In this study, Whitmania Pigra Whitman (WPW), Hirudo nipponica Whitman (HNW) and Whitmania acranutata Whitman (WAW) were hydrolyzed by biomimetic enzymatic hydrolysis to obtain the active peptides of WPW (APP), the active peptides of HNW (APH) and the active peptides of WAW (APA), respectively. Then their structures were characterized by sykam amino acid analyzer, fourier transform infrared spectrometer (FT-IR), circular dichroism (CD) spectrometer and LC-MS. Next, the anti-thrombotic activities of APP, APH and APA were determined by carrageenan-induced tail vein thrombosis model in mice, and the anti-thrombotic mechanisms of high-dose APP group (HAPP), high-dose APH group (HAPH) and high-dose APA group (HAPA) were explored based on UHPLC-Q-Exactive Orbitrap mass spectrometry.Results: The results showed that the amino acid composition of APP, APH and APA was consistent, and the proportion of each amino acid was few different. The results of FT-IR and CD showed that there were no significant differences in the proportion of secondary structures (such as β-sheet and random coil) and infrared absorption peaks between APP, APH and APA. Mass spectrometry data showed that there were 43 common peptides in APP, APH and APA, indicating that the three have common material basis. APP, APH and APA could significantly inhibit platelet aggregation, reduce black-tail length, whole blood viscosity (WBV), plasma viscosity (PV), and Fibrinogen (FIB), and prolong coagulation time, including activated partial thrombin time (APTT), prothrombin time (PT) and thrombin time (TT). In addition, 24 metabolites were identified as potential biomarkers associated with thrombosis development. Among these, 19, 23, and 20 metabolites were significantly normalized after administration of HAPP, HAPH, and HAPA in the mice, respectively. Furthermore, the intervention mechanism of HAPP, HAPH and HAPA on tail vein thrombosis mainly involved in linoleic acid metabolism, primary bile acid biosynthesis and ether lipid metabolism.Conclusion: Our findings suggest that APP, APH and APA can exert their anti-blood stasis and anti-thrombotic activities by interfering with disordered metabolic pathways in vivo, and there is no significant difference in their efficacies.

show abstract

Section: Discussionmentioning

confidence: 99%

Characterization of active peptides derived from three leeches and comparison of their anti-thrombotic mechanisms using the tail vein thrombosis model in mice and metabonomics

Dong,

Li,

et al. 2024

Front. Pharmacol.

View full text Add to dashboard Cite

show abstract

“…In addition to hirudin, leeches have many antithrombotic agents in their bodies and secreted saliva. For example, bdellin, similar to hirudin, showed anticoagulant effects by prolonging the activated partial thromboplastin time (Cheng et al 2019). Destabilase (Zavalova et al 1996), leech carboxypeptidase inhibitor (LCI) (Reverter et al 2000), and hyaluronidase (Jin et al 2014) were suggested to have thrombolytic properties.…”

Section: Introductionmentioning

confidence: 99%

Comparative genomics of three non-hematophagous leeches (Whitmaniaspp.): focusing on antithrombotic genes

Zhao,

Huang,

Tang

et al. 2024

Preprint

View full text Add to dashboard Cite

Leeches are well known for their blood-feeding habits and are widely used for medicinal purposes as they secrete various antithrombotic substances. However, some leeches such as Whitmania spp. exhibit non-hematophagous feeding habits and their significance for medicinal use is debated. In this study, we provide chromosome-level genomes of two non-hematophagous leeches Whitmania acranulata and Whitmania laevis, and combined with our previous results of Whitmania pigra, we systematically analyzed the similarities and differences on the genomes and especially their antithrombotic genes among the three non-hematophagous Whitmania leeches. For W. acranulata, W. laevis, and W. pigra, the genome size (181.72 Mb, 173.87 Mb, and 173.56), the percentage of repeat sites (29.55%, 28.28%, and 27.02%), and the number of protein-coding genes (27,068, 23,805, and 24,156) were close to each other, respectively. In contrast, both the total number of the antithrombotic genes (100, 63, and 79), and the detailed constitutes of different antithrombotic gene families were obviously different among the three leeches. There were also massive genetic variations among the members within each antithrombotic gene/protein family. RNA-Seq-based gene expression estimation showed that the expression profiles of the antithrombotic gene families were apparently different among the three leeches. This is the most comprehensive comparison of the genomes and antithrombic biomacromolecules for the Whitmania leeches to date. Our results will greatly facilitate the evolutionary research and application of leech derivatives for medical and pharmaceutical purposes of thrombosis.

show abstract

“…In combination with hirudin, leeches possess numerous antithrombotic bioactive compounds within their bodies and secreted saliva. For instance, bdellin, like hirudin, lengthened activated partial thromboplastin time and exerted anticoagulant capability [26]. Destabilase [27], leech carboxypeptidase inhibitor [28], and hyaluronidase [29] were believed to have thrombolytic properties.…”

Section: Introductionmentioning

confidence: 99%

Revisiting the Asian Buffalo Leech (Hirudinaria manillensis) Genome: Focus on Antithrombotic Genes and Their Corresponding Proteins

Liu,

Zhao,

Huang

et al. 2023

Genes

View full text Add to dashboard Cite

Leeches are well-known annelids due to their obligate blood-feeding habits. Some leech species secrete various biologically active substances which have important medical and pharmaceutical value in antithrombotic treatments. In this study, we provided a high-quality genome of the Asian buffalo leech (Hirudinaria manillensis), based on which we performed a systematic identification of potential antithrombotic genes and their corresponding proteins. Combining automatic and manual prediction, we identified 21 antithrombotic gene families including fourteen coagulation inhibitors, three platelet aggregation inhibitors, three fibrinolysis enhancers, and one tissue penetration enhancer. A total of 72 antithrombotic genes, including two pseudogenes, were identified, including most of their corresponding proteins forming three or more disulfide bonds. Three protein families (LDTI, antistasin, and granulin) had internal tandem repeats containing 6, 10, and 12 conserved cysteines, respectively. We also measured the anticoagulant activities of the five identified hirudins (hirudin_Hman1 ~ hirudin_Hman5). The results showed that three (hirudin_Hman1, hirudin_Hman2, and hirudin_Hman5), but not the remaining two, exhibited anticoagulant activities. Our study provides the most comprehensive collection of antithrombotic biomacromolecules from a leech to date. These results will greatly facilitate the research and application of leech derivatives for medical and pharmaceutical purposes in the treatment of thrombotic diseases.

show abstract

Purification and characterization of a novel anti-coagulant from the leech Hirudinaria manillensis

Cited by 11 publications

References 30 publications

Characterization of active peptides derived from three leeches and comparison of their anti-thrombotic mechanisms using the tail vein thrombosis model in mice and metabonomics

Characterization of active peptides derived from three leeches and comparison of their anti-thrombotic mechanisms using the tail vein thrombosis model in mice and metabonomics

Comparative genomics of three non-hematophagous leeches (Whitmaniaspp.): focusing on antithrombotic genes

Revisiting the Asian Buffalo Leech (Hirudinaria manillensis) Genome: Focus on Antithrombotic Genes and Their Corresponding Proteins

Contact Info

Product

Resources

About