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Indian Journal of Pharmacology 1998; 30: 16-20 RESEARCH PAPER

ANTI-CATARACT ACTIVITY OF OCIMUM SANCTUM ON EXPERIMENTAL


CATARACT

P. SHARMA, S. KULSHRESHTHA, A.L. SHARMA

Department of Pharmacology, S.N. Medical College, Agra - 282 001

Manuscript Received: 17.4.1997 Revised: 13.9.1997 Accepted: 26.10.1997

SUMMARY Objective: To study the effect of Ocimum sanctum (Tulsi) on experimental cataract in rats and rabbits.
Methods: Two models of experimental cataract were induced: (1) Galactosaemic cataract in rats by
30% galactose, (2) Naphthalene cataract in rabbits by 1 gm/kg naphthalene. Ocimum sanctum (O.S.)
was administered orally in both models at two dose levels 1 and 2 gm/kg of body weight for curative
and prophylactic effects. The study was conducted for 40 days.
Results: O.S. delayed the onset of cataract as well as the subsequent maturation of cataract significantly
in both models. In addition to delay in reaching various stages of development of cataract, IV stage
did not develop with high doses till completion of 40 days of experimental period.
Conclusion: O.S. delayed the process of cataractogenesis in both models. The higher doses are
more effective and have aot promising prophylactic role rather than curative one. This effect is more
clear in galactosaemic c&r&t.

KEY WORDS Ageing cataract Ocimum sanctum galactose naphthalene adaptogens

INTRODUCTION antiemetic, diaphoretic, expectorant and is also used


for catarrhal bronchitis, bronchial asthma, dysentery
Ageing is a mysterious physiological process and
and various skin diseasesgslO. It has been shown
leads to a gradual decline in antioxidant enzymesl.
to be effective as antistress and adap to enic and
Endogenous anti-oxidative enzymes constitute a tis-
attenuates the stress induced changes Bl,12.
sue defence system against ageing and stress, likely
to be caused by free radicals2. These enzymes
MATERIALS AND METHODS
like glutathione reductase and peroxidase and super
oxide dismutase are necessary for removing the Young Charles Foster rats (2-4 weeks age) weighing
free radicals from the system and the lens also. 100-l 25 gm and albino rabbits (l-l .25 kgs) of either
Senile cataract is an a e related problem and a sex, free from ocular pathology, were obtained from
major cause of blindness B .There are about 12 million Veterinary College, Mathura. Animals were housed
blind people due to cataract in India alone4. There in light (12 hours light - 12 hours dark cycle) and
are many causative factors for cataract formation temperature (23*2OC) controlled conditions. They
other than age-like nutritional deficiencies, environ- were fed on standard pellet diet (Lipton, India) and
mental changes, radiation, metabolic diseases like given water ad libitum.
diabetes and oxidative stres@. The oxidation of cel-
lular and membrane constituents occurs along with Plan of study
the cataractogenic degeneration of lens and an- Animals were randomly grouped and sub-grouped
tioxidative enzyme levels in lens are decreased2e617. having six animals each and were given catarac-
For arresting this age related cataract, although a togenic challenge and test drug (Table 1).
single drug is not available, certain substances have Induction of cataract
been reported. Some plant products and drugs are
reported to have anti-stress-adaptogenic activity. Galactosaemic cataract (GC) was induced in rats
Keeping in account the role of free radicals and by keeping them on diet containing 30% galactose
oxidative damage in induction of cataract8 adap- up to 40 days13. Naphthalene cataract (NC) was
togen, O.S. was selected for the present study. OS. induced in rabbits by administering naphthalene
also known as Tulsi has been used as analgesic, (lgm/kg) in liquid paraffin orally’4.
ANTI-CATARACT ACTIVITY OF OCIMUM SANCTUM 17

Table 1. Plan of study period. “Z” test was applied to know the significance
Group Subgroup Model of Dose (gm/kg)
of preventive effect. In some animals appearance
(n=6) Cataract of O.S./Saline of cataract and its progression from first stage to
subsequent stages was delayed or even arrested.
Normal - - -
Student’s t-test was applied to calculate the sig-
Control la - G Saline nificance of delaying effect on the development of
experimental cataract in all groups.
II II-C-A G 1
II-C-B G 2
RESULTS
Ill III-P-A G 1

Ill-P-B G 2 Normal
Rats and rabbits kept under standard conditions
Control lb - N Saline
and diet did not elicit any abnormality in the eyes
IV IV-C-A N 1 or lens during study.
IV-C-B N 2 Cataract control
V V-P-A N 1 Saline treated control rats subjected to galactosae-
mic challenge showed 100% response (Table 2).
V-P-A N 2
Similarly all rabbits subjected to naphthalene
C = Curative group. O.S. started from the first day of challenge developed cataract (Table 3).
cataractogenic challenge.
P = Prophylactic group. OS. started 10 days prior to the Effect of O.S. in galactosaemic cataract
cataractogenic challenge. (Table 2)
G = Galactosaemic cataract; N = Naphthalene cataract.
Curative group
O.S. significantly delayed the appearance as well
Drug as its progression to various stages with both doses
Fresh leaves of O.S. were crushed in concentration of 1 and 2 gm/kg. Mature cataract was not developed
of 1 gm/5 ml of distilled water at room temperature. in any rat.
It was administered orally at two dose levels of
1 gm/kg and 2 gm/kg body weight in both curative Prophylactic group
and prophylactic groups. O.S. was found to be highly effective when it was
administered 10 days prior to cataractogenic
Examination of eyes challenge. Lenticular opacity appeared after a sig-
Eyes were examined daily by oblique illumination nificant delay and its progress to further stages was
and direct ophthalmoscopy for noting changes in also affected, especially with 2 gm/kg dose. Stage
lens. Slit lamp examination was done as and when III and IV were not observed and cataract progressed
required till the end of experimental period. Initiation only to stage II.
and progression of lenticular opacity was graded
into four stages15. First stage-invasion of lens by Effect of O.S. in naphthalene cataract (Table 3)
vacuoles on periphery. Second stage-presence of
linear opacities in naphthalene cataract; structural Curative group
and vacuolar opacities in centre of lens in galac- In this group at both the dose levels O.S. inhibited
tosaemic cataract. Third stage-presence of plaques progression of cataract as evidenced by significant
and clouding of lens; and Fourth stage-mature cata- delay in reaching various stages of cataractogene-
ract. sis.
Statistical analysis Prophylactic group
Observations showed two types of effects of O.S. Mature cataract did not appear in any animal. Len-
In some animals development of experimental cata- ticular opacity was arrested at stage II only in high
ract was prevented completely and even first stage dose. Results were found to be statistically signifi-
of cataract could not appear during 40 days of study cant for both initiation and progression of cataract.
18 P. SHARMA, S. KULSHRESHTHA AND A.L. SHARMA

Table 2. Effect of Ocimum sanctum in galactosaemic cataract Table 3. Effect of Ocimum sanctum in naphthalene cataract

Group Stage Rats Developing Day (Mean&D) Group Stage Rabb Developing Day (Mean&D)
(n=6) of No. cataract of appearance (n=6) of -its cataract of appearance of
cataract % of cataract cataract No. % cataract

la I 6 100.00 5.90 f 0.55 lb I 6 100.00 4.58 i 0.48

II 6 100.00 10.55 zt 0.63 II 6 100.00 6.20 * 0.69

Ill 6 100.00 23.98 i 0.89 Ill 6 100.00 8.50 f 0.96

IV 6 100.00 33.13 f 0.52 IV 6 100.00 11.38 i 0.82

II-C-A I 4 66.67 15.66 f 0.43a IV-C-A I 4 66.67 10.00 f 0.6Ea

II 3 50.00’ 22.50 i 0.75’ II 4 66.67 18.80 f 0.90a

III 2 33.33” 34.06 * 0.86’ Ill 3 50.00’ 24.85 f 0.28a

IV 1 16.67” 40.00a IV 2 33.33” 34.23 f 0.34a

II-C-B I 3 50.00 18.34 f 0.76a IV-C-B I 4 66.67 15.80 zt 0.65a

II 2 33.33” 29.00 f 0.8Ea II 2 33.33” 26.00 i 0.56a

Ill 1 16.67” 36.00a Ill 2 33.33” 35.00 zt 0.38a

IV 0 0 Not appeared IV 0 0 Not appeared

III-P-A I 2 33.33** 24.00 i 0.58a V-P-A I 4 66.67 19.00 i 0.52a

II 2 33.33** 32.80 zt 0.46a II 2 33.33” 25.00 i 0.64a

Ill 1 16.67** 38.52a Ill 1 16.67”. 37.33a

IV 0 0 Not appeared IV 0 0 Not appeared

Ill-P-B I 2 33.33** 30.46 i 0.98a V-P-B I 3 50.00’ 27.50 zt 0.78a

II 2 33.33” 38.50 f 0.50a II 2 33.33” 39.00 f 0.4ga

Ill 0 0 Not appeared Ill 0 0 Not appeared

IV 0 0 Not appeared IV 0 0 Not appeared

fi = 1 gm/kg; B = 2gm/kg. A = 1 gm/kg; B = 2gm/kg.


P < 0.05; P < 0.01 when compared to control (Z-test) *P c 0.05; **P < 0.01 when compared to control (Z-test)
aP < 0.001 when compared to control (t-test). aP < 0.001 when compared to control (t-test).

DISCUSSION antioxidants in preventing various diseases and in


Cataract is a major cause of blindness all over the human cataract formation is reported’89’g. The free
worldss4. It is an age related phenomenon, over radicals are the main causative factors in cataract.
and above oxidative stress also plays its role. Sur- The toxic species of oxygen formed in lens milieu
gical treatment has remained the only remedy till include superoxide anion, lipid hydroperoxides, OH
now. Hence, if a drug is sought which can either radicals and hydrogen peroxide. Endogeneous an-
reverse or prevent lenticular opacity, it will be a tioxidant like reduced glutathione is present in high
great advance in the treatment of this disorder. A concentration in lens, super-oxide dismutase and
number of drugs have been shown to interfere with catalase keep the level of free radicals below toxic
the process of cataract formation like aldose levels. In cataractous lenses its concentration is
reductase inhibitors, restatin, sulindac, aspirin, decreased2’. Hence, with the use of antioxidants
quercetin15-17. The potential role of vitamins and cataract formation can be prevented. O.S. has been
ANTI-CATARACT ACTIVITY OF OClMUM SANCTUM 19

reported to increase physical endurance and body 3. Sasaki K, Karinok, Kajima M. Cataract survey in the local
area using photographic documentation. Dev Ophthalmol
resistance in animals subjected to various types 1987;15:28-36.
of physical and chemical stress and normalizes bio-
logical and histopathological changes produced by 4. Minassian DC, Mehra V. 3.6 million blinded by cataract
stress, showing that it possesses significant anti- each year. Projections from the first epidemiological study
stress properties. Stressful conditions are known of incidence of cataract blindness in India. BrJ Ophthalmol
1990;74:341-3.
to increase the susceptibility of an individual to vari-
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and immunosuppression”. Erythrocyte membrane for cataract. Eye 1987;1:537-41.
is taken as an experimental model to study bio-
membrane. Restraint stress produces changes in 6. Kobayashi S, Kasuya M, Shimizu K, Takehana M, Sakai
K, Suzuki N, ltoni M. Glutathione isoprophylester (YM 737)
the structure of erythrocyte membrane. There is inhibits the progression of X-ray induced cataract in
increase in protein clusterization, increase in fluidity rats.Curr Ey e Res 1993;12:115-22.
and reduction in thickness. The resultant effect is
an increase in cell permeability. O.S. attenuates 7. Reddy VN, Giblin FJ, Matsuda H. Defence system of the
all these stress induced changes to normal*‘. Stress lens against oxidative damage. In: Srivastava, (ed.) Red
Blood Cell and Lens Metabolism. 1980:139-54.
besides effecting other organs like causing gastric
ulcer and immunosuppression, might also produce a. Kador PF, Kinoshita JH. Diabetic and galactosaemic cata-
changes in lenticular membranes and permeability. racts. Ciba Fdn Symp 1984;6:110-31.
It is therefore concluded that OS. can delay as 9. Singh N, Nath R, Gupta ML, Bhargava KP. An expeimental
well as arrest the progress of cataractogenesis. The evaluation of anti-asthmatic potentialities of lnula racemosa.
effect is more with higher doses. It has got promising Quart J Crude Drug Res 1980;18:86-96.
prophylactic role and is more clear in galactosaemic
cataract which is more close to diabetic cataract. 10. Bharvaga KP, Singh N. Anti-stress activity of Ocimum sanc-
O.S. is known to possess adaptogenic and antistress tum. Indian J Med Res 1981;73:443-51.
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11. Singh N, Verma P, Mishra N, Nath R. A comparative evalu-
suspension are reported to possess significant im- ation of some antistress agents of plant origin. lndian J
munostimulant activity in rats23124. Prevention of Pharmacol 1991;23:99-103.
cataract may be throu h a mechanism involving
free radical scavenging%5 , preventing lipid peroxi- 12. Singh N, Mishra N, Srivastava AK, Dixit KS, Gupta GP.
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may be actin as anticataract by inhibiting lipid per- 13. Yudkin AM. Cataract produced in albino rats on a ration
oxidation27p2 B. Antioxidants viz. vitamin “C” and “E”*s containing a high proportion of lactose or galactose. Arch
p-carotene are also acting through free radical scav- Ophthal 1935;14:960-7.
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poglycaemic activity30. This may help in arresting 14. Salmony, Dorrit. Some biochemical changes in naphthalene
cataract. Br J Ophthal 1960;44:29-33.
diabetic cataract process. It is hereby concluded
that daily consumption of O.S. may delay emergence 15. Sharma P, Sharma AL, Kulshreshtha S, Mishra SS, Pandey
of lenticular opacity. However, further studies are DN. An expertmental study of quercetin in lenticular opaci-
needed for biochemical parameters. ties. Indian J Pharmacol 1982;14:169-75.

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f
INDIAN PHARMACOLOGICAL SOCIETY AWARDS - 1997

ACHARI PRIZE Pallabi De, S.C. Dasgupta and A. Gomes


Lab. of Toxinology and Experimental Pharmacodynamics,
Department of Physiology, University of Calcutta, Calcutta.

N.N. DUTTA PRIZE D. Kumar, H.C. Tripathi, S.K. Mishra,S.K. Tandon, V. Ravi Prakash
and S.C. Mishra
Division of Pharmacology and Toxicology, Division of Standardization,
I.V.R.I., lzatnagar - 243 122, (U.P.)

P.C. DANDIYA PRIZE L.K. Gupta, M. Choudhury, S. Khetarpal and G. Tayal


Department of Pharmacology, Lady Hardinge Medical College,
New Delhi.

P.P. SURYAKUMARI Ashish R. Sevak and Ramesh K. Goyal


PRIZE Department of Pharmacology, L.M. College of Pharmacy,
P.O.Box No.4011, Navrangpura, Ahmedabad - 380 009.

UVNAS PRIZE Doodipale S. Reddy, Manjeet Singh, Sujata Ghosh and N.K. Ganguly
Department of Pharmacology, University Institute of Pharmaceutical
Sciences, Punjab University, Chandigarh - 160 014.

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