Metabolite profiling and mechanisms of bioactivity of snake autolysate - A traditional Uzbek medicine

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Abstract

Ethnopharmacological relevance

Aqueous autolysate from the snake Eryx miliaris (SNA) has been used in traditional medicine of Uzbekistan as anti-inflammatory, hepatoprotective and immunomodulatory agent. However, little is known about the chemical composition and its mechanisms of activity.

Aim of the study

This is our first attempt to analyse the composition of snake autolysate using gas chromatography with mass spectrometry (GC-MS) and to investigate the mechanisms of anti-inflammatory and hyaluronidase activity of fingerprinted E. miliaris autolysate to support their use in the traditional Uzbek medicine.

Materials and methods

Aqueous autolysate was evaporated and derivatised for GC-MS analysis of metabolites. For quantification, lipids were extracted from autolysate by solvent extraction and derivatised by esterification and silylation. Biological activity was evaluated with lipid peroxidation, cyclooxygenase (COX) inhibition and antihyaluronidase activity tests.

Results

GC-MS analysis of SNA enabled the identification of 27 compounds. Short chain fatty acids (SCFA, 21%), amino acid/derivatives 39% (incl. 2-piperidinone 19%), phenyl (7%), and OH-Phenyl (10%) derivatives covered 77%. Other derivatives (9%) included succinic acid and 3-indole acetic acid). Long chain fatty acids (C16–C18) accounted for 3%. The lipid concentration of SNA was 1.2 mg/mL (0.12%). Three concentration levels (1.0–20.0 μg/mL) did not inhibit COX-1 and COX-2 in vitro and malondialdehyde level was not decreased by SNA in lipid peroxidation model. However, SNA was a potent inhibitor of the hyaluronidase enzyme activity in a dose dependent manner with IC50 = 0.086 mL/mL.

Conclusion

The results from GC-MS analyses of SNA lead us to the identification of a wide range of major chemical structures of the metabolites and their derivatives with several categories. Pharmacological studies support the traditional use of SNA and show one of its possible mechanisms of activity via inhibition of hyaluronidase.

Introduction

The snake used to be a permanent companion of healers and magicians from very ancient times and early references to snakes used medicinally are known since Greek and Roman writings. Already in the 18th century, the belief in viper flesh as a medicinal remedy was known by European physicians, who prescribed it for many disorders like snakebite, tumors and childbirth pains (McDonald, 1994). More recently, according to the interviewers, snake products have been used to treat 21 different diseases in Brazilian traditional medicine (Alves and Filho, 2007; Alves, 2009). The snake preparations are used in Russian officinal medicine (Prokopov et al., 2019). Fat and whole coral snakes Micrurus sp., whole tiger snake Spilotes pullatus and whole parrot snake Leptophis ahetula are recommended for the treatment of rheumatism and as pain relief caused by sting of animals (Alves and Rosa, 2007).

Eryx miliaris Pall., known commonly as the Dwarf sand boa (Uzbek name Ko'lvor ilani - blind snake), is a species of snake in the Boidae family. The species is endemic to Iran, Afghanistan, Turkmenistan, Uzbekistan, southern Russia, and northwestern China (McDiarmid et al., 1999). The whole snake E. miliaris without removing any part and cooked as a soup has been used against asthma in Uzbekistan (Sezik et al., 2004). Aqueous autolysate from the whole snake E. miliaris, known as ‘Erixin’ (or ‘Reptilin’) is used in the traditional medicine of Uzbekistan as anti-inflammatory, hepatoprotective and immunomodulatory drug. It has been found to be effective in rheumatological diseases, tuberculosis, brucellosis, acute obstructive or chronic bronchitis and dermatosclerosis (Ubaidullaev et al., 1999; Silantiyeva et al., 2000; Akbarov et al., 2003; Asatova et al., 2003).

The aim of the study was: i) to analyse the chemical composition of snake autolysate (SNA) of E. miliaris by gas chromatography-mass spectrometry (GC-MS) for the first time, and ii) to investigate possible mechanisms of anti-inflammatory activity of fingerprinted SNA to support their use in the traditional Uzbek medicine.

Section snippets

Snake sample

The snake autolysate (SNA) of E. miliaris was received from “NPC Sv-Bio” (Uzbekistan), batch# 10.2017. According to manufacturer information, the whole snake E. miliaris after euthanasia in CO2 camera were located in closed vessels, and left for autolysis in thermostatic condition; the resulted product was filtered and sterilised.

Derivatisation of autolysate samples for GC-MS analysis

The liquid snake autolysate sample was evaporated to dryness, and the residue was trimethylsilylated at 100 °C. In addition, the autolysate sample was extracted for

Results and discussion

In this study the chemical composition of SNA was analysed by GC-MS. The metabolic profiling provides an insight of the chemical compositions of SNA (Fig. S1, Table S1), which contained 27 identified compounds (Fig. 1). Short chain fatty acids (SCFA, 21%), amino acid/derivatives 39% (incl. 2-piperidinone 19%), phenyl (7%), and OH-Phenyl (10%) derivatives covered 77% of identified compounds. Other derivatives (9%) included succinic acid and 3-indole acetic acid. In autolysate, long chain fatty

Conclusions

The results from GC-MS analyses of SNA lead us to the identification of a wide range of major chemical structures of the metabolites and their derivatives with several categories. Pharmacological studies support the traditional use of SNA and show one of its possible mechanisms of activity via inhibition of hyaluronidase.

Declaration of competing interest

The authors declare no conflicts of interests.

References (34)

  • R.R.N. Alves

    Fauna used in popular medicine in Northeast Brazil

    J Ethnobiol Ethnomed

    (2009)
  • R.R.N. Alves et al.

    Commercialization and use of snakes in North and Northeastern Brazil: implications for conservation and management

    Vertebrate Conservation and Biodiversity

    (2007)
  • T. Aoki et al.

    Prostaglandins in chronic inflammation, 3-17

  • S.S. Asatova et al.

    Hepatoprotector properties of eryxin in animals with acute drug-induced hepatitis model

    Pharm. Chem. J.

    (2003)
  • B. Dalile et al.

    The role of short-chain fatty acids in microbiota-gut-brain communication

    Nat. Rev. Gastroenterol. Hepatol.

    (2019)
  • F.N. Djakhangirov et al.

    Experimental study of some pharmacological properties of eryxin preparation

    Infect. Immun. Pharmacol.

    (2000)
  • Y. Ji et al.

    Indole-3-acetic acid alleviates nonalcoholic fatty liver disease in mice via attenuation of hepatic lipogenesis, and oxidative and inflammatory stress

    Nutrients

    (2019)
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