Preprint / Version 1

In silico, in vitro, and in vivo acute and sub-acute toxicity profiling of whole plant methanol extract of Equisetum diffusum D. Don from the sub-Himalayan West Bengal, India, having ethnobotanical uses

Authors

  • Sourav Sarkar Department of Zoology, Cell and Molecular Biology Laboratory, University of North Bengal, Raja Rammohunpur, Darjeeling, West Bengal 734013 India
  • Debabrata Modak Department of Zoology, Cell and Molecular Biology Laboratory, University of North Bengal, Raja Rammohunpur, Darjeeling, West Bengal 734013 India
  • Sudipta Roy Department of Zoology, Cell and Molecular Biology Laboratory, University of North Bengal, Raja Rammohunpur, Darjeeling, West Bengal 734013 India
  • Anupam Biswas Chemical Science and Technology Division, Pharmacology Lab, Natural Products Chemistry Group, CSIR-North East Institute of Science and Technology (CSIR-NEIST), Jorhat, Assam 785006 India
  • Mafidul Islam Department of Biotechnology, Gauhati University, Guwahati, Assam 781014 India
  • Rinku Baishya Chemical Science and Technology Division, Pharmacology Lab, Natural Products Chemistry Group, CSIR-North East Institute of Science and Technology (CSIR-NEIST), Jorhat, Assam 785006 India
  • Sujoy Bose Department of Biotechnology, Gauhati University, Guwahati, Assam 781014 India
  • John Georrge Department of Bioinformatics, University of North Bengal, Darjeeling, West Bengal 734013 India
  • Soumen Bhattacharjee Department of Zoology, Cell and Molecular Biology Laboratory, University of North Bengal, Raja Rammohunpur, Darjeeling, West Bengal 734013 India

Keywords:

Equisetum diffusum D. Don, ADME-Toxicity, HEK293, Huh7, Acute toxicity, Sub-acute toxicity

Abstract

Equisetum diffusum D. Don commonly known as ‘Himalayan horsetail’, has been traditionally used in the treatment of back pain, bone fracture and dislocation, and arthritis by various tribal communities of India. Our previous study confirmed the anti-inflammatory efficacy of the plant through in silico, in vitro, and in vivo model studies. Therefore, the current research is focused on safety dose evaluation for the first-time of the whole-plant methanol extract (EDME) of E. diffusum through appropriate in silico, in vitro, and in vivo approaches. Method The whole plant, along with its rhizomes, was collected, and the methanol extract was prepared. The in silico ADMET study was performed to predict the pharmacokinetics profile and toxicity of all the identified phyto-compounds of EDME previously screened by GC–MS study. In vitro cytotoxicity study of EDME was performed using two cell lines: kidney (HEK293) and liver (Huh7) cell lines. The in vivo toxicity study of EDME was validated by the acute toxicity (OECD 423, 2002) and sub-acute toxicity assays (OECD 407, 2008) in the Wistar Albino rat model. Results The in silico ADMET study of all 47 bioactives predicted good pharmacokinetic and low toxicity profiles. In vitro cytotoxicity showed higher IC50 values of EDME viz., 672 ± 15.7 μg/mL and 1698 ± 6.54 μg/mL for both kidney (HEK293) and liver (Huh7) cell lines, respectively, which were considered as low-toxic. Based on acute oral toxicity, the LD50 value of the extract was considered “non-toxic” up to a feeding range of 2000 mg/kg of body weight. The regular consumption of the extract for an extended period (28 days) was also qualified as safe based on the body and organ weight, hematological, biochemical, and histoarchitecture results in the sub-acute toxicity assay. Conclusion The detailed in silico, in vitro, in vivo (acute and sub-acute oral toxicity) studies gave us a new insight to the safety dose evaluation of Equisetum diffusum, which may serve as a reliable documentation for undertaking the experimental validation of the ethnobotanical uses of the plant which would help in the field of drug development for the treatment of inflammation related complications. Supplementary Information The online version contains supplementary material available at 10.1186/s12906-024-04606-y. Keywords: Equisetum diffusum D. Don, ADME-Toxicity, HEK293, Huh7, Acute toxicity, Sub-acute toxicity