Pharmaceutics Drug Interaction between Clove Oil (Eugenia caryophyllata) and Chloroform in mice

Objective: Anaesthetics are compounds that produce loss of sensation and muscle relaxation. They may cause loss of consciousness. Many compounds have been used for anaesthesia, Clove oil is a anesthetic compound. The results obtained by GC/MS analysis showed that clove essential oil contains 36 components. The highest concentration was eugenol (6). Eugenol is widely applied as anesthetics, analgesics, anti-inflammatory agents, and flavoring agents. In recent years, the pharmacodynamics of eugenol has been developed to immunological function, central nervous regulation, reproductive effects, cardiovascular system, digestive system, blood biochemistry, and urinary system. Results obtained from human studies also confirmed analgesic effects of the plant in toothache and in patients suffering from anal fissure (3,7). Chloroform is the other compound that use for anesthesia in laboratory studies. Induction of anesthesia by chloroform is very fast. However, due to its hepatotoxicity side effects its used is limited to laboratory animals. The aim of this study was to investigate the pharmaceutical drug intraction between clove oil (Eugenia caryophyllata) and chloroform. Materials and Methods: This study was approved by the Ethics Committee of Shahrekord University. About 30 adult female mice with body weight ranged from 27-32 g were used in the present study. They kept in cages under conditions of controlled temperature and 12h day-night cycle for 2 weeks, allowed free access to water and fed ad libitum. Mince were randomly divided into two groups of 15 animals each. Group 1 which received only normal saline throughout the course of the experiment was used as control. In group 2, clove oil was given orally for 45 consecutive days at the dose of 40 mg kg-1. Different concentrations of Essential oil were prepared in 10 ml saline. At the end of this period, mice were weighted again, and provided a quiet non-stressful environment. Then, mice were anesthetized using chloroform. Time to the onset and duration of the anesthesia were recorded. Time duration between placing mice in desiccator and anaesthetized was considered induction time or onset time, Time from anaesthetizing to recovery was considered as duration time of anesthesia. All samples were analyzed and the results were assessed statistically using t test. P values less than 0.05 were considered statistically significant. All calculations were performed using Graph pad Prism 5 software. Results and Discussion: The onset time of anesthesia with chloroform in group2 was significantly (p 0.05) longer (155±13 sec) than group1 (55±5 sec).The duration time in group2 was significantly (p 0.05) lesser (137±7sec) than group1 (345±26 sec). The present study is the first to investigate the pharmaceutical drug interaction between Clove Oil (Eugenia caryophyllata) and Chloroform. The result of present study showed that the onset time of anesthesia was 55±5 sec with chloroform in control group (group1), increasing to 155±13 sec in group 2 which was given clove oil for 45 days. However, the duration time of anesthesia was 345±26 sec in group1, decreasing to 137±7sec in group2. The mechanism of anesthetic action of eugenol on fish first starts with the inhibition of the cerebral cortex (period of tactile loss), followed by the basal ganglion and cerebellum (excitation period), and finally the spinal cord (narcosis stage). Eugenol is widely applied to current local nerve anesthesia. Muller et al. studied the adjustment of eugenol to the excitability of sciatic nerve of mouse. They found that eugenol mainly inhibits Na+-dependent channel receptor, activates transient receptor potential vanilloid receptor (TRPV1) subfamily of ion channel receptors, and inhibits the flow of sodium ions, thus, blocking nerve conduction and decreasing the conduction rate gradually (5). Low eugenol content can inhibit complex action potential and decrease nerve excitability. Eugenol produces anesthesia in rodents similar to propofol (4), and alleviates thermal hypersensitivity in an experimental model of neuropathic pain in rats (4). Eugenol inhibits N-methyl-D-aspartate (NMDA) receptors but potentiates ionotropic γ- aminobutyric acid (GABAA) receptors, which are both involved in pain sensitivity (1). Eugenol depresses compound action potentials in both A and C fibers which may explain its analgesic effects. Eugenol is similar in chemical structure to capsaicin and therefore its effect on a vanilloid receptor should not be ignored (8). It was also shown that eugenol inhibits Na+ currents in rat dorsal root ganglion neurons (2). Induction of anesthesia by chloroform is very fast. The mechanism of anesthetic action of chloroform is to inhibit K+ currents. Therefore, there is may be an interaction in ion channel functions. Conclusion: The result of present study showed the existence of a pharmaceutical drug interaction between clove oil and chloroform. Further work should perform to find out the exact mechanism of this interaction.