The saliva of the medicinal leech Hirudo medicinalis—1. Biochemical characterization of the high molecular weight fraction

doi:10.1016/0305-0491(87)90053-8

Comparative Biochemistry and Physiology Part B: Comparative Biochemistry

Volume 87, Issue 3, 1987, Pages 567-573

https://doi.org/10.1016/0305-0491(87)90053-8 Get rights and content

Abstract

1.
1. A method is described for obtaining dilute Hirudo medicinalis saliva by feeding leeches through a membrane on arginine/saline and squeezing them immediately after from the posterior end forwards. The process can be repeated at intervals. Yields are considerably higher than from salivary gland extracts.
2.
2. Hirudo saliva contains hirudin, eglin, hyaluronidase, collagenase and apyrase. Leech collagenase and apyrase are here reported for the first time.
3.
3. On gel filtration of lyophilized saliva, activity peaks were well defined. Approximate molecular weights were determined. Apyrase appears in two forms with optimum activity around pH 7.5. Collagenase was identified as belonging to the mammalian type.

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2024, Pharmacological Research - Modern Chinese Medicine
Leech extract contains many identified bioactive substances which have a variety of biological effects. Leech extract was discovered in the late 19th century and since then many pharmaceutical products have been produced using leech extract for different ailments, but many have been withdrawn. Ongoing studies focus on developing health authority compliant pharmaceuticals to be used as modern medicine.
The online databases including Google Scholar, Scopus, PubMed, and Web of Science, were searched using different keywords: "Leech", "Leech extract", "Leech salivary gland extract" and "Leech saliva extract". The purpose of this review was to discuss the development of leech extract as a therapeutic agent, including the evolution of extraction techniques, and the successful manufacturing of leech extract-based pharmaceuticals.
Leech extract has been successfully developed as a therapeutic agent. Some of the developed leech extract-based pharmaceutics were withdrawn and some are still in the market. The extraction methods played a vital role in the quality and efficacy of leech extract-based pharmaceutics and ultimately on their sustainability in the market. Also, the full characterization of leech extract components is a key factor in the development of leech extract as a therapeutic agent.
This review provides a comprehensive historical perspective on the development of leech extract therapy, exploring its various stages of development and the key scientific and medical advances that have shaped its current state. The discussion also delves into potential future applications.
Leech extract is an invaluable source of bioactive substances that can be utilized for the treatment of mild and life-threatening medical disorders. This review will encourage other scientists to continue their research on leech extract, especially in the areas of formulation and marketing.
In-depth profiles of bioactive large molecules in saliva secretions of leeches determined by combining salivary gland proteome and transcriptome data
2019, Journal of Proteomics
Citation Excerpt :
Leeches were starved for three weeks. Saliva was obtained by a method described in the literature [30]. Saliva proteins and peptides were denatured with lysis buffer (8 M urea and protease inhibitor cocktail) and then applied to an ultrafiltration tube with a 3-kD cut-off.
Medicinal leeches have been widely utilized in medical procedures for thousands of years. The application of leeches depends on the components of leech saliva secretions and active molecules, but many components of the secretions are not well characterized due to their low concentration and abundance. Determination of the profiles of leech salivary secretions is important to its medicinal application. In this study, we performed an in-depth proteomic analysis of leech salivary glands and deduced 434 full-length protein sequences from combined leech proteome and transcriptome databases. After integrating data from both datasets, forty-four proteins and two hundred twenty-one transcripts of bioactive molecules were involved in leech sucking pathways. Using gene expression analysis, we found that two-thirds of bioactive genes played key roles in leech bite processes and were associated cave-dwelling habitats. Our results indicate that the treatment efficiency can differ depending on the sucking leech species. Moreover, combining high-throughput proteomic and transcriptomic analyses is effective for the determination of wide profiles of proteins that are present at low concentrations in secretions. These findings highlight the extensive diversity of bioactive molecules and provide a new foundation for performing novel investigations and discovering future pharmacological agents or targets in leech medicinal therapy.
Medicinal leech therapy has been used for many centuries depending on the components of leech saliva secretions and active actions, but many components of the secretions were less known due to its low concentration and abundance. Determination of the profiles of leech salivary secretions is important to its medicinal application. Hereby, the molecular information provided by proteomic and transcriptomic analysis can be used to develop a more thorough understanding of leech sucking pathway and medicinal application. It provided a new foundation for performing novel investigations and discovering future pharmacological agents or targets in leech medicinal therapy.
Leech therapy in reconstructive maxillofacial surgery
2012, Journal of Oral and Maxillofacial Surgery
Corrective plastic surgery is indicated after accidents, burns, cancer surgery, or postoperative wound healing disorders with large tissue defects. The range of reconstructive techniques includes local skin flaps, pedicle grafts, and microvascular anastomosed flaps in the case of large defects. Main complications such as arterial and venous insufficiency caused by a vessel collapse or a vascular spasm are reported regularly in the area of anastomosed vessels and are the concern of any surgeon. Today, leeches are used if wound healing is at risk because of hemodynamic imbalance or a venous insufficiency.
A retrospective evaluation of 148 patients who underwent medical leech therapy in the case of local or pedicaled flaps and some patients who had undergone reconstruction with microvascular flaps from 2005 and 2010 was conducted. Our sample had the typical symptoms of venous congestions of their flaps, despite suture removal, relief of pressure on the flap, and the elimination of a hematoma beyond the flap after surgery. Medical leech therapy was used in these cases.
Our series has confirmed the excellent and predictable healing after medical leech therapy for local and microsurgical anastomosed flaps in the case of venous congestion.
Leech therapy should be considered as a reliable additional procedure and an advantage in maxillofacial and plastic reconstructive surgery to remedy complications resulting from a hemodynamic imbalance or venous insufficiency in the immediate postoperative period.
Elimination of symbiotic Aeromonas spp. from the intestinal tract of the medicinal leech, Hirudo medicinalis, using ciprofloxacin feeding
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Markwardt [2] isolated and characterized this molecule and in 1986 it was produced by genetic engineering [3]. In recent years, recognition of a number of antithrombotic substances and a vasodilator in the saliva of the medicinal leech [4,5] has generated renewed interest in the use of the leech itself to promote venous drainage in tissues whose vitality is threatened by venous congestion and obstruction, especially after plastic and reconstructive surgery. In 2004, this treatment modality received the approval of the Food and Drug Administration in the USA.
The use of the medicinal leech (Hirudo medicinalis) in promoting venous drainage in tissues whose vitality is threatened by venous congestion and obstruction, especially in plastic and reconstructive surgery, has been complicated by infections caused by Aeromonas spp. These are leech endosymbionts for which patients undergoing hirudotherapy frequently receive systemic chemoprophylaxis. In order to evaluate the possibility of rendering leeches safe for use on patients, H. medicinalis were fed artificially with a 2 g/L arginine solution (used as a phagostimulant) supplemented with ciprofloxacin (100 mg/L). Aeromonads were detected in 57 out of 80 control leeches (71.3%), but in none of the 56 leeches treated with ciprofloxacin (p <0.001). Treated leeches survived for up to 4 months. Tested weekly, 61% of these leeches took human blood for at least 4 weeks after treatment and all remained negative for aeromonads. All water samples in which leeches were kept before treatment were contaminated with Aeromonas spp.; none were detected in any of the NaCl/arginine solutions with which treated animals were fed. Molecular characterization of two phenotypically distinct isolates using gyrB sequencing showed that one clustered tightly with A. veronii and the other was closely related to A. media. Other environmental bacteria and fungi were isolated from 26.5% of treated leeches that had taken a blood meal 1–4 weeks after treatment. Ciprofloxacin reduced the number of leech-associated aeromonads to undetectable levels for extended periods. Most treated leeches were ready to take a blood meal after treatment, suggesting the possibility of using ciprofloxacin-treated leeches instead of chemoprophylaxis in patients undergoing hirudotherapy.
Effectiveness of leech therapy in women with symptomatic arthrosis of the first carpometacarpal joint: A randomized controlled trial
2008, Pain
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Different mechanisms may explain the observed effects. First, a variety of pharmacological active substances besides hirudin have been described in leech saliva, such as histamin-like vasodilators, kallikrein and tryptase inhibitors, a variety of other proteinase inhibitors and anaesthetics [3,26,27,32]. Through the concomitant activity of a further leech saliva component, hyaluronidase [4] these substances might reach deeper tissue zones, and, possibly, the joint space.
Leech therapy has been shown to be effective for symptomatic treatment of osteoarthritis of the knee. We aimed to investigate the effectiveness of leech therapy in another type of osteoarthritis, osteoarthritis of the first carpometacarpal joint (thumb saddle joint). Thirty-two women with symptomatic painful osteoarthritis of the first carpometacarpal joint and who scored > 40 mm on a 100 mm VAS pain scale were randomized to a single treatment with 2–3 locally applied leeches (leech group) or a 30-day course with topical diclofenac twice a day. Primary outcome measure was change of overall pain (mean of VAS for pain at rest, in motion, during grip) from baseline to day 7. Secondary outcomes were functional disability (DASH-questionnaire), quality of life (QoL, SF-36) and grip strength. Patients were examined baseline and at days 7, 30 and 60 after treatment. Overall pain score at day 7 was reduced from 59.6 ± 13.8 to 27.1 ± 20.6 in the leech group (n = 16) and from 50.6 ± 13.3 to 46.9 ± 18.5 with diclofenac (n = 16) (group difference −26.5, 95%CI −40.3; −12.7; p = 0.0003). Group differences for pain relief favoring the leech treatment increased at days 30 and 60. Significant treatment effects were also observed for the DASH score, QoL and grip. Results were not affected by outcome expectation or consumption of analgetics. A single course of leech therapy is effective in relieving pain, improving disability and QoL for at least 2 months. The potential of leech therapy for treatment of arthritic pain and underlying mechanisms should be further investigated.
Effects of broodstock density and diet on reproduction and juvenile culture of the Leech, Hirudinaria manillensis Lesson, 1842
2008, Aquaculture
Citation Excerpt :
However, in more recent years, the medically beneficial usage of leech is once again increasing (Baskova et al., 1983, 1992). For example, H. medicinalis is used by plastic surgeons to restore venous circulation in tissue grafts where blood stagnation is a problem (Sawyer, 1986; Rigbi et al., 1987; Roters and Zebe, 1992; Whitaker et al., 2004; Huang et al., 2006b). In order to meet the demand from clinical use, Chinese traditional medicine and other scientific research, there has been growing interest in culturing and breeding leeches in many countries (Yang, 1996; Trontelj and Utevsky, 2005).
The leech Hirudinaria manillensis, Lesson 1842 is of much interest for clinical and medicinal use. In this study, the effect of broodstock density (5, 10, 15, 20, 25, 30 and 50 ind tank^− 1) on the reproductive efficiency of H. manillensis was examined. After 4 months of culture, the number of cocoons produced was significantly different among the different broodstock densities (F_{6, 34} = 4.560, P < 0.05), but fertilization ratio was not (F_{6, 34} = 1.319, P = 0.285). The average number of hatchlings per cocoon (5.72 ± 0.13 ind) and hatching rate (96.82 ± 1.31%) of the cocoons in the 5 ind tank^− 1 treatment were significantly higher than those of the other treatments. The 50 ind tank^− 1 treatment had the highest mortality of parent leeches (29.60 ± 2.48%). The size and wet weight of the cocoons were significantly different among the treatments, with the 5 ind tank^− 1 treatment having the largest cocoon size (standard length and diameter) and wet weight. The time of juvenile release from the cocoons did not differ significantly among the different broodstock densities (P > 0.05).
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General reviewThe saliva of the medicinal leech Hirudo medicinalis—1. Biochemical characterization of the high molecular weight fraction

Abstract

Br. J. Plast. Surg.

Archs Biochem. Biophys.

Comp. Biochem. Physiol.

J. biol. Chem.

Leeches (Hirudinea)

Leeches (Hirudinea)

Cell

J. biol. Chem.

Experientia

J. exp. Med.

Biochim. biophys. Acta

Hoppe-Seyler's Z. physiol. Chem.

J. biol. Chem.

Br. J. Plast. Surg.

Effect of the salivary gland secretion of Hirudo medicinalis on the extrinsic and intrinsic mechanisms of blood clotting

Byul. Eksp. Biol. Meditsin.

Salivary gland secretion of the leech Hirudo medicinalis inhibits ADP-induced human platelet adhesion on a collagen-coated surface

Byul. Eksp. Biol. Meditsin.

Antithrombin, antitrypsin and antichymotrypsin activities of the salivary gland secretion and intestinal chyme of medicinal leeches

General review
The saliva of the medicinal leech Hirudo medicinalis—1. Biochemical characterization of the high molecular weight fraction