Albizia lebbeck seed methanolic extract as a complementary therapy to manage local toxicity of Echis carinatus venom in a murine model

Abstract

Context and objective: Viperid venom-induced chronic local-toxicity continues even after anti-snake venom treatment. Therefore, traditional antidote Albizia lebbeck L. (Fabaceae) seed extract was tested against Echis carinatus S. (Viperidae) venom (ECV)-induced local toxicity to evaluate its complementary remedy.

Materials and methods: Soxhlet extraction of A. lebbeck seeds was performed with the increasing polarity of solvents (n-hexane to water); the extract was screened for phytochemicals (alkaloids, anthraquinones, flavonoids, glycosides, phenolics, saponins, steroids and tannins). In preliminary in vitro analysis, A. lebbeck methanolic extract (ALME) demonstrated significant inhibition of ECV proteases, the major enzyme–toxin responsible for local- toxicity. Therefore, in vitro neutralizing potential of ALME was further evaluated against hyaluronidases and phospholipase A₂ (1:1–1:100 w/w). In addition, alleviation of ECV induced characteristic local- toxicity [haemorrhage (i.d.) and myotoxicity (i.m.)] was determined in mice.

Results: ALME contained high concentrations of phenolics and flavonoids and demonstrated significant in vitro inhibition of ECV protease (IC₅₀=36.32 μg, p < 0.0001) and hyaluronidase (IC₅₀=91.95 μg, p < 0.0001) at 1:100 w/w. ALME significantly neutralized ECV induced haemorrhage (ED₅₀=26.37 μg, p < 0.0001) and myotoxicity by significantly reducing serum creatinine kinase (ED₅₀=37.5 μg, p < 0.0001) and lactate dehydrogenase (ED₅₀=31.44 μg, p = 0.0021) levels at 1:50 w/w.

Discussion and conclusion: ALME demonstrated significant neutralization of ECV enzymes that contribute in local tissue damage and haemostatic alterations. The study scientifically supports the anecdotal use of A. lebbeck in complementary medicine and identifies ALME as principle fraction responsible for antivenom properties.

Keywords:

• Folk medicine
• haemorrhage
• hyaluronidases
• myotoxicity
• proteases
• saw scaled viper
• woman’s tongue tree

Acknowledgements

Authors thank Institutional Animal and Human Ethical Committees (IAEC and IHEC) for ethical clearance. Authors thank Dr. K. A. Sharvani, herbarium in-charge, Yuvaraja's College, UOM for plant identification. Authors also thank G. Swamy, M. S. Sumanth, M. N. Savitha and G. V. Rudresha for their support during the work.

Disclosure statement

The authors have no conflicts of interests to disclose.

Funding information

Authors thank University Grants Commission, New Delhi, and Institution of Excellence (funded by Ministry of Human Resource Development, Government of India), University of Mysore (UOM), Mysuru for Fellowship. Authors thank UGC-SAP, DST-PURSE and VGST for financial assistance to DOS in Biochemistry, UOM and Vijnana Bhavan, UOM.