scientist , department of crop and herbal physiology , jawaharlal nehru krishi vishwa vidyalaya , jabalpur ( m.p ) . hd / chpy/980 was deposited at the jawaharlal nehru krishi vishwavidyalaya ( jnkvv ) , jabalpur ( m.p . ) . m. oleifera was dried in shade , coarsely powered and used for preparation of extract . the powder was extracted with ethanol : water ( 1:1 ) by stirring for 48 h and filtered through whatman no . the residue was then extracted with two additional 200 ml portions of ethanol : water ( 1:1 ) as described above . the combined extracts were then evaporated ( buchi type , york science c. , mumbai ) at 40c to dryness , and stored at 4c for further use . potassium chloride , sodium chloride , sodium bicarbonate , sodium phosphate , and calcium chloride were purchased from central drug house ( cdh ) , india ; glucose was purchased from fischer scientific ( india ) , trichloroacetic acid , and ethylenediaminetetraacetic acid ( edta ) were purchased from qualigens , india ; thiobarbituric acid was purchased from sigma , us . fresh goat eyeballs were obtained from slaughterhouse immediately after slaughter and transported to the laboratory at 0 - 4c . the lenses were removed by extra capsular extraction and incubated in artificial aqueous humor ( nacl 140 mm , kcl 5 mm , mgcl 2 2 mm , nahco3 0.5 mm , nah ( po4 ) 2 0.5 mm , cacl2 0.4 mm and glucose 5.5 mm ) at room temperature and ph 7.8 for 72 h. penicillin 32 mg % and streptomycin 250 mg % were added to the culture media to prevent bacterial contamination . glucose in a concentration of 55 mm was used to induce cataract . at high concentrations , glucose in the lens was metabolized through sorbitol pathway and accumulation of polyols ( sugar alcohols ) , causing over hydration and oxidative stress . extract was dissolved in dimethyl sulfoxide ( dmso ) and used for the study . in present study , a dose of 250 g / ml and 500 g / ml of extract of m. oleifera were used . a total of 24 lenses were divided into following categories ( n = 6 in each category ) : group i : normal lens [ control ( glucose 5.5 mm ) ] group ii : glucose 55 mm group iii : glucose 55 mm + extract 250 g / ml group iv : glucose 55 mm + extract 500 g / ml the protein content of the samples was determined by the method of lowry et al . , using bovine serum albumin as the standard . the lipid peroxide formed was estimated by measuring thiobarbituric acid reacting substances ( tbars ) . the groups were compared using one - way anova followed by bonferroni 's test using glucose 55 mm group as control . scientist , department of crop and herbal physiology , jawaharlal nehru krishi vishwa vidyalaya , jabalpur ( m.p ) . hd / chpy/980 was deposited at the jawaharlal nehru krishi vishwavidyalaya ( jnkvv ) , jabalpur ( m.p . ) . m. oleifera was dried in shade , coarsely powered and used for preparation of extract . the powder was extracted with ethanol : water ( 1:1 ) by stirring for 48 h and filtered through whatman no . the residue was then extracted with two additional 200 ml portions of ethanol : water ( 1:1 ) as described above . the combined extracts were then evaporated ( buchi type , york science c. , mumbai ) at 40c to dryness , and stored at 4c for further use . potassium chloride , sodium chloride , sodium bicarbonate , sodium phosphate , and calcium chloride were purchased from central drug house ( cdh ) , india ; glucose was purchased from fischer scientific ( india ) , trichloroacetic acid , and ethylenediaminetetraacetic acid ( edta ) were purchased from qualigens , india ; thiobarbituric acid was purchased from sigma , us . fresh goat eyeballs were obtained from slaughterhouse immediately after slaughter and transported to the laboratory at 0 - 4c . the lenses were removed by extra capsular extraction and incubated in artificial aqueous humor ( nacl 140 mm , kcl 5 mm , mgcl 2 2 mm , nahco3 0.5 mm , nah ( po4 ) 2 0.5 mm , cacl2 0.4 mm and glucose 5.5 mm ) at room temperature and ph 7.8 for 72 h. penicillin 32 mg % and streptomycin 250 mg % were added to the culture media to prevent bacterial contamination . glucose in a concentration of 55 mm was used to induce cataract . at high concentrations , glucose in the lens was metabolized through sorbitol pathway and accumulation of polyols ( sugar alcohols ) , causing over hydration and oxidative stress . in present study , a dose of 250 g / ml and 500 g / ml of extract of m. oleifera were used . a total of 24 lenses were divided into following categories ( n = 6 in each category ) : group i : normal lens [ control ( glucose 5.5 mm ) ] group ii : glucose 55 mm group iii : glucose 55 mm + extract 250 g / ml group iv : glucose 55 mm + extract 500 g / ml the protein content of the samples was determined by the method of lowry et al . , using bovine serum albumin as the standard . the lipid peroxide formed was estimated by measuring thiobarbituric acid reacting substances ( tbars ) . the groups were compared using one - way anova followed by bonferroni 's test using glucose 55 mm group as control . where p < 0.05 was considered significant . phytochemical studies of m. oleifera extract revealed the presence of glycosides , alkaloids , phenolics and flavonoids . the lens protein level of glucose ( 55 m ) treated lens ( group ii ) showed significant decrease ( p < 0.005 ) as compared to normal control group ( group i ) m. oleifera extract at the concentration of 250 g / ml and 500 g / ml showed significant increase ( p < 0.05 ) in lens protein as compared to toxic control ( group ii ) [ fig . 1 ] . effects of m.oleifera extract on total protein level on isolated goat lens ; all values are mean  sd , n = 6 ; statistical comparison was performed using analysis of variance ( anova ) followed by bonferronis test ( * p < 0.05 ) the lens glutathione level of glucose ( 55 m ) treated lens ( group ii ) showed significant decrease ( p < 0.05 ) as compared to normal control group ( group i ) m. oleifera extract at the concentration of 250g / ml and 500 g / ml showed significant increase ( p < 0.05 ) in lens glutathione as compared to toxic control ( group ii ) . only treated group ( group iv ) showed almost the same levels of glutathione as that of control [ fig . 2 ] . effects of m.oleifera extract on glutathione level on isolated goat lens ; all values are mean  sd , n = 6 ; statistical comparison was performed using analysis of variance ( anova ) followed by bonferronis test ( * p < 0.05 ) in this study , incubation by sugars resulted in a time dependent inactivation of the enzymes . lens catalase activities were also significantly lower in the lens of group ii as compared to extract treated groups ( group iii and iv ; p < 0.05 ) [ fig . effects of m.oleifera extract on catalase level on isolated goat lens ; all values are mean  sd , n = 6 ; statistical comparison was performed using analysis of variance ( anova ) followed by bonferronis test ( * p < 0.05 ) mda levels were found to be very high in glucose 55 mm treated lens compared with normal lens . lens treated with m. oleifera extract had significantly reduced mda content ( p < 0.05 ) at both concentrations compared with high glucose group [ fig . 4 ] . effects of m.oleifera extract on mda level on isolated goat lens ; all values are mean  sd , n = 6 ; statistical comparison was performed using analysis of variance ( anova ) followed by bonferronis test ( * p < 0.05 ) lenses incubated in glucose 5.5 m remained transparent , whereas , the lens incubated in 55 m glucose developed dense opacities . the opacity increased towards centre with complete opacification at the end of 72 h. incorporation of m. oleifera extract ( 250 g / ml and 500 g / ml ) retarded the development of opacity [ fig . ( a ) group i : normal lens after 72 hours of incubation in glucose 5.5 mm ( transparency maintained , squares clearly visible ) . ( b ) group ii : complete cataractogenesis after 72 hours of incubation in glucose 55 mm ( absolute loss of transparency , no squares visible through lens ) . ( c ) group iii : after 72 hours of incubation in glucose 55 mm + extract 250 g / ml , lens appears slightly hazy ( very less no . of squares slightly visible ) . ( d ) group iv : after 72 hours of incubation in glucose 55 mm + extract 500 g / ml , lens appears slightly hazy ( more no . of squares visible ) phytochemical studies of m. oleifera extract revealed the presence of glycosides , alkaloids , phenolics and flavonoids . the lens protein level of glucose ( 55 m ) treated lens ( group ii ) showed significant decrease ( p < 0.005 ) as compared to normal control group ( group i ) m. oleifera extract at the concentration of 250 g / ml and 500 g / ml showed significant increase ( p < 0.05 ) in lens protein as compared to toxic control ( group ii ) [ fig . 1 ] . effects of m.oleifera extract on total protein level on isolated goat lens ; all values are mean  sd , n = 6 ; statistical comparison was performed using analysis of variance ( anova ) followed by bonferronis test ( * p < 0.05 ) the lens glutathione level of glucose ( 55 m ) treated lens ( group ii ) showed significant decrease ( p < 0.05 ) as compared to normal control group ( group i ) m. oleifera extract at the concentration of 250g / ml and 500 g / ml showed significant increase ( p < 0.05 ) in lens glutathione as compared to toxic control ( group ii ) . only treated group ( group iv ) showed almost the same levels of glutathione as that of control [ fig . 2 ] . effects of m.oleifera extract on glutathione level on isolated goat lens ; all values are mean  sd , n = 6 ; statistical comparison was performed using analysis of variance ( anova ) followed by bonferronis test ( * p < 0.05 ) in this study , incubation by sugars resulted in a time dependent inactivation of the enzymes . lens catalase activities were also significantly lower in the lens of group ii as compared to extract treated groups ( group iii and iv ; p < 0.05 ) [ fig . effects of m.oleifera extract on catalase level on isolated goat lens ; all values are mean  sd , n = 6 ; statistical comparison was performed using analysis of variance ( anova ) followed by bonferronis test ( * p < 0.05 ) mda levels were found to be very high in glucose 55 mm treated lens compared with normal lens . lens treated with m. oleifera extract had significantly reduced mda content ( p < 0.05 ) at both concentrations compared with high glucose group [ fig . 4 ] . effects of m.oleifera extract on mda level on isolated goat lens ; all values are mean  sd , n = 6 ; statistical comparison was performed using analysis of variance ( anova ) followed by bonferronis test ( * p < 0.05 ) the lens glutathione level of glucose ( 55 m ) treated lens ( group ii ) showed significant decrease ( p < 0.05 ) as compared to normal control group ( group i ) m. oleifera extract at the concentration of 250g / ml and 500 g / ml showed significant increase ( p < 0.05 ) in lens glutathione as compared to toxic control ( group ii ) . only treated group ( group iv ) showed almost the same levels of glutathione as that of control [ fig . effects of m.oleifera extract on glutathione level on isolated goat lens ; all values are mean  sd , n = 6 ; statistical comparison was performed using analysis of variance ( anova ) followed by bonferronis test ( * p < 0.05 ) in this study , incubation by sugars resulted in a time dependent inactivation of the enzymes . lens catalase activities were also significantly lower in the lens of group ii as compared to extract treated groups ( group iii and iv ; p < 0.05 ) [ fig . 3 ] . effects of m.oleifera extract on catalase level on isolated goat lens ; all values are mean  sd , n = 6 ; statistical comparison was performed using analysis of variance ( anova ) followed by bonferronis test ( * p < 0.05 ) mda levels were found to be very high in glucose 55 mm treated lens compared with normal lens . lens treated with m. oleifera extract had significantly reduced mda content ( p < 0.05 ) at both concentrations compared with high glucose group [ fig . 4 ] . effects of m.oleifera extract on mda level on isolated goat lens ; all values are mean  sd , n = 6 ; statistical comparison was performed using analysis of variance ( anova ) followed by bonferronis test ( * p < 0.05 ) lenses incubated in glucose 5.5 m remained transparent , whereas , the lens incubated in 55 m glucose developed dense opacities . the opacity increased towards centre with complete opacification at the end of 72 h. incorporation of m. oleifera extract ( 250 g / ml and 500 g / ml ) retarded the development of opacity [ fig . ( a ) group i : normal lens after 72 hours of incubation in glucose 5.5 mm ( transparency maintained , squares clearly visible ) . ( b ) group ii : complete cataractogenesis after 72 hours of incubation in glucose 55 mm ( absolute loss of transparency , no squares visible through lens ) . ( c ) group iii : after 72 hours of incubation in glucose 55 mm + extract 250 g / ml , lens appears slightly hazy ( very less no . of squares slightly visible ) . ( d ) group iv : after 72 hours of incubation in glucose 55 mm + extract 500 g / ml , lens appears slightly hazy ( more no . of squares visible ) cataract is the most prevalent disorder leading to visual impairment , the prospect of pharmacological intervention to inhibit or to delay the onset of cataract is still at the experimental stage . cat , sod , and gsh are important components of the innate enzymatic defenses of the lens . cat has been shown to be responsible for the detoxification of significant amounts of h2o2 sod catalyzes the removal of superoxide radicals ( o ) , which would otherwise damage the membrane and biological structures . the enzyme gsh , first demonstrated in the lens by pirie , has been reported to maintain the integrity of the phospholipid bilayer of membranes by inhibiting lipid peroxidation . cat and gsh catalyze the transformation of h2o2 within the cell to harmless by products , thereby curtailing the quantity of cellular destruction inflicted by products of lipid peroxidation . a reduction in the activities of these enzymes in tissues has been associated with the accumulation of highly reactive free radicals , leading to deleterious effects such as loss of integrity and function of cell membranes . phytoconstituents from herbal drugs may ultimately inhibit expenditure of gsh through oxidation leaving the  sh groups intact . on the other hand , they may directly stimulate reduced glutathione synthesis which may be due to a modulating effect on reduced glutathione related enzymes in the lens . the restoration of reduced glutathione levels by m. oleifera extract also warranted its anticataract potential . during oxidative stress , denaturation and aggregation of lens proteins the situation is however improved along with an increase in the protein level in lens treated with m.oleifera . the preventive role of extract has also been demonstrated by the restoration of protein . in this study the levels of mda were more in group ii when compared with group i , iii and iv which suggest preventive role against cataract . in vitro anti - cataract effect of m. oleifera on isolated goat lenses incubated in a high glucose medium was dose - dependent . hence it can be conclude that oxidative stress is an important factor in the development of glucose induced cataract and the use of anti - oxidants may be advocated in patients to delay or prevent formation of cataract . in conclusion m. oleifera showed promising in vitro activity against glucose cataract in an isolated goat lens model . result of study is encouraging , but further studies are required to access the use of m. oleifera in humans for prevention of cataract .
aim of study : the aim of current work was to evaluate in vitro anticataract potential of moringa oliefera extract.materials and methods : goat eye lenses were divided into 4 groups ; group served as control , group ii as toxic control , group iii and group iv were incubated in extract ( 250 g / ml and 500 g / ml of extract of m. oliefera ) group ii , iii and iv were incubated in 55 mm glucose in artificial aqueous humor to induce lens opacification . 
 estimation of total , water soluble protein , catalase , glutathione and malondialdehyde along with photographic evaluation of lens was done.results:group ii ( toxic control ) lenses showed high amount of mda ( malondialdehyde ) , soluble , insoluble protein , decreased catalase and glutathione levels , while lenses treated with moringa oliefera extract ( group iii and group iv ) showed significant ( * p < 0.05 ) reduction in mda and increased level of catalase , glutathione , total and soluble protein.conclusion:results of present findings suggest protective effect of moringa oliefera in prevention of in vitro glucose induced cataract .