Effect of Methyl Chavicol on the Candida albicans Intracellular Glutathione

Abstract

Herbs, have long been recognized for their medicinal values. Ocimum sanctum belongs to the family Labiatae and is known as Tulsi in Hindi and Holy Basil in English. Tulsi constituents are chiefly femchol, methyl chavicol, linalool, eugenol and ß-caryophyllene, responsible for its anti-bacterial, anti-septic and anti-fungal action.  Inhibitory effects of aqueous and alcoholic extracts of Tulsi have been observed on enteric pathogens by Agar diffusion method. Tulsi is also known to have numerous pharmacological activities, such as antidiabetic, antistress, immunomodulatory, analgesic, antipyretic, anti inflammatory, antiulcerogenic, antihypertensive, CNS depressant, hepatoprotective, radioprotective, antitumor and antimicrobial activities. Methyl chavicol a major constituent of Holy Basil (Ocimum sanctum) essential oil has been evaluated against the effect of methyl chavicol on the intracellular Glutathione. Cytosolic reduced glutathione was determined by the method of Jollow with slight modifications. In this method 1.0 ml of the cell free extract was precipitate with 1.0 ml of 4 % sulphosalicylic acid. The samples were then kept for 1 hour at 4ºC and centrifuged at 1200 g for 15 minutes at 40ºC. The assay mixture consisted of 0.1 ml of above supernatant, 2.7 ml of Phosphate buffer (0.01 M, pH 7.4) and 0.2 ml of freshly prepared 5, 5’- dithiobis-2-nitrobenzene (DTNB) in a total volume of 3.0 ml. The colour developed due to the formation of a yellow colored complex, 5-thio-2-nitrobenzoate, was measured immediately at 412 nm at 25ºC. It has been found in our study that the levels of glutathione were found to be 144μmole/gm at zero hours i.e. in yeast uninduced cells. Post 2 hours induction shows that levels of reduced glutathione in control untreated cells were found to be 124.4375μmole/gm cells. It went down as the induction period increases; at the end of 24hours induction the GSH levels were found to be 86.56775μmole/gm. These results in agreement with the previous studies.  All treated cells showed decreased levels of GSH as the induction period increases. Decrease in GSH was found to be in dose dependent manner. A subsequent fall in the value of reduced glutathione was observed with the increasing concentrations of MC with the lowest being 28.452μmole/gm cells at 50µg/ml concentration of MC after 24 hrs inducton. This result is in coordination with the assumption that methyl chavicol may also affect the glutathione metabolic enzymes and may have affected the cell defense armory.