Ocimum sanctum Modulates Selenite-Induced Cataractogenic Changes and Prevents Rat Lens Opacification

Purpose: To study the effect of Ocimum sanctum (OS) on selenite-induced morphological and biochemical changes in isolated rat lenses as well as on cataract incidence in rat pups. Methods: Transparent rat lenses were divided into normal, selenite-only, and four treated groups. Selenite-only and treated group lenses were subjected to oxidative stress in vitro by incorporating sodium selenite (100 μ M) in the culture medium. The effect of OS (70, 140, 280, and 560 μ g/ml) was studied on the levels of reduced glutathione (GSH) and thiobarbituric acid reacting substances (TBARS) in selenite-challenged lenses. The lowest concentration of OS offering significant modulation on these two parameters was determined. Subsequently, the effect of prior and cotreatment with the lowest effective concentration of OS was studied on TBARS, GSH, and on lens antioxidant enzymes such as superoxide dismutase (SOD), glutathione peroxidase (GSHPx), catalase (CAT), and glutathione-S-transferase (GST). Changes in lens protein profiles under different incubation conditions were analyzed by SDS gel-electrophoresis. In vivo, cataract was induced by a single subcutaneous injection of sodium selenite (25 μ mole/kg b.w.) to 9-day-old rat pups. The anticataract effect of OS (5 and 10 mg/kg b.w.) injected intraperitoneally 4 hr prior to selenite challenge was evaluated by the presence of lens nuclear opacity in rat pups on the 16th postnatal day. Insolubilization of lens proteins post–selenite injection was monitored for 4 days. Results: The lenses in the selenite-only group developed cortical opacities in 24 hr. OS showed different degrees of positive modulation in selenite-induced morphological as well as biochemical changes. The lowest effective dose of OS that significantly modulated glutathione and thiobarbituric acid reacting substances was found to be 140 μ g/ml. At this dose, a significant increase in antioxidant enzyme levels and preservation of normal lens protein profile was observed. OS at the dose of 70 μ g/ml did not show any significant protection with respect to either morphology or biochemistry of lenses. In vivo, 5 and 10 mg/kg of OS reduced the incidence of selenite cataract by 20% and 60%, respectively, and prevented protein insolubilization as well. Conclusions: Aqueous extract of OS possesses potential anticataract activity against selenite-induced experimental cataractogenesis. The protective effect was supported by restoration of the antioxidant defense system and inhibition of protein insolubilization of rat lenses as well.

[1]  J. Harding,et al.  Free and protein-bound glutathione in normal and cataractous human lenses. , 1970, The Biochemical journal.

[2]  W B Jakoby,et al.  Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. , 1974, The Journal of biological chemistry.

[3]  H. Sies,et al.  Oxidative stress: oxidants and antioxidants , 1997, Experimental physiology.

[4]  Arun Sharma,et al.  A Handbook of Medicinal Plants: A Complete Source Book , 2004 .

[5]  P. Uma Devi Radioprotective, anticarcinogenic and antioxidant properties of the Indian holy basil, Ocimum sanctum (Tulasi). , 2001, Indian journal of experimental biology.

[6]  P. U. Devi,et al.  Modulation of glutathione and antioxidant enzymes by Ocimum sanctum and its role in protection against radiation injury. , 1999, Indian journal of experimental biology.

[7]  T. Shearer,et al.  Calpain inhibitor, SJA6017, reduces the rate of formation of selenite cataract in rats , 2001, Current eye research.

[8]  J. Prakash,et al.  Chemopreventive activity of Ocimum sanctum seed oil. , 2000, Journal of ethnopharmacology.

[9]  John I. Clark,et al.  Effect of selected anti-cataract agents on opacification in the selenite cataract model. , 1996, Experimental eye research.

[10]  I. Fridovich,et al.  The oxidation of phenylhydrazine: superoxide and mechanism. , 1976, Biochemistry.

[11]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[12]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[13]  Sujata Joshi,et al.  Lycopene attenuates oxidative stress induced experimental cataract development: an in vitro and in vivo study. , 2003, Nutrition.

[14]  S. Srivastava,et al.  Green Tea (Camellia sinensis) Protects against Selenite-Induced Oxidative Stress in Experimental Cataractogenesis , 2002, Ophthalmic Research.

[15]  R. Saxena,et al.  Epidemiology of Cataract in India: Combating Plans and Strategies , 1999, Ophthalmic Research.

[16]  T. Shearer,et al.  Selenite nuclear cataract: review of the model. , 1997, Molecular vision.

[17]  H. Gerster Antioxidant vitamins in cataract prevention , 1989, Zeitschrift fur Ernahrungswissenschaft.

[18]  S. Gupta,et al.  Role of naproxen as anti-oxidant in selenite cataract. , 1994, Ophthalmic research.

[19]  W. Valentine,et al.  Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. , 1967, The Journal of laboratory and clinical medicine.

[20]  G. Sahin,et al.  Effects of some probable antioxidants on selenite-induced cataract formation and oxidative stress-related parameters in rats. , 1999, Toxicology.

[21]  P. Uma Devi,et al.  Radiation Protection by the Ocimum Flavonoids Orientin and Vicenin: Mechanisms of Action , 2000, Radiation research.

[22]  S. Sood,et al.  Protective effect of Tulsi (Ocimum Sanctum) on lipid peroxidation in stress induced by anemic hypoxia in rabbits. , 2003, Indian journal of physiology and pharmacology.

[23]  A. Spector,et al.  Oxidative stress‐induced cataract: mechanism of action , 1995, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[24]  S. Varma,et al.  Attenuation of sugar cataract by ethyl pyruvate , 1999, Molecular and Cellular Biochemistry.

[25]  K. Yagi,et al.  Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. , 1979, Analytical biochemistry.

[26]  B. Mannervik,et al.  Levels of glutathione, glutathione reductase and glutathione S-transferase activities in rat lung and liver. , 1979, Biochimica et biophysica acta.