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all al. (2012)
Research Article
56 | International Journal of Life Sciences • ISSN 2091–0525 • Year 2012 • Volume 6 • Issue 1
Research Article Deepa et al. (2012)
of transition metals as well as generation of ROS history of steroid intake, other ophthalmic disease,
during the process of glycation. Oxidative stress also and systemic disorders were excluded from the study.
affects the expression of antioxidant enzymes by its The study was approved by the Institutional Time
action on signal transduction. Hyperglycemia can also Bound Research Committee. A written informed
cause inactivation of the existing enzymes by consent was taken from the subjects. Postoperatively
glycation. Thus oxidative stress may develop by cataractous lens were collected in ice cold 0.9%
insufficient antioxidant activity even if ROS Normal saline container and transferred in ice box to
production is within physiological range (King et al., the place of analysis and analyzed on the same day
1996) immediately for the following parameters: For
determining the antioxidant activity and calcium
Calcium is an important extracellular cation in levels in lens, the lens was homogenized with 1.2ml
the body, total calcium in the body is about 1-1.5kg. ice cold 0.01M phosphate buffer pH 7.4. Mixture was
99% is in bone and 1% in extracellular fluid. It is centrifuged at 2500rpm for 10mins. 100µl
present both in extracellular and intracellular supernatant was used for analysis. The total lens
compartments. The cell membrane is generally antioxidant activity was determined by Koracevic’s
impermeable to calcium ions. Normally the calcium method (Koracevic et al., 2001) the assay measures
levels are maintained at low concentration inside the the capacity of antioxidant to inhibit the production of
cell. This is maintained by Na+ K+ exchanger; Na+ Ca2+ thiobarbituric acid reactive substances from sodium
exchanger and calcium ATPase pump (Galvan, 1988). benzoate under the influence of the oxygen free
Optimum intracellular calcium is necessary to radicals derived from Fenton reaction. The reaction
maintain lens transparency. Calcium is maintained in was measured spectrophotometrically at 535nm.
micromoles range in the cytoplasm by regulatory Calcium in the lens was determined by
systems. In normal lens, the ionized calcium is less Cresolphthalein complexone method (Stern, 1957)
than 1% of the total concentration and below 0.1% of using calcium kits from Aspen Lab Pvt Ltd.
the level in surrounding aqueous humor (Tomlinson Cresolphthalein complexone a metal complex dye
et al., 1991). reacts with calcium ions in alkaline medium forming a
purple color. Intensity of the color formed is directly
Metal ions are known to play an essential role
proportional to the calcium concentration and was
in living systems, both in growth and in metabolism.
measured photometrically at 570nm. Since we
Impaired metabolism of trace elements is observed in
analyzed in lens tissue we need to express the levels
diabetic patients. Alteration in the capacity to
of total antioxidant activity and calcium in milligrams
maintain normal calcium homeostasis have been
of proteins .So protein content was determined by the
suggested to underlie the reduced cellular functions
Lowry’s method (Lowry et al., 1951) blue color
characteristic of aging process and predisposes the
obtained was measured spectrophotometrically using
senescent organism to diverse pathologies viz
Bovine albumin as standard
Cancer, Heart disease, Neurodegenerative disease
including Cataract (Thomas et al., 2000; Ernesto et Statistical Analysis
al., 1987) Altered calcium homeostasis due to
oxidative stress leads to increased intracellular The data was statistical analysis by epi-info
calcium levels and in turn leads to cataract formation software using Analysis of Variance [ANOVA]. Since
(Wride, 1996; Nakamura et al., 2000) two way ANOVA showed that the interaction between
cataract and maturity is significant, the comparison
The aim of the study was to evaluate the total
between the type of cataract and maturity was done
antioxidant activity and calcium levels in diabetic
separately using independent ‘t’ test. The results were
patients lens depending on type of cataract and to
expressed as Mean ± Standard deviation.
correlate the levels of calcium with the antioxidant
activity in progression of cataract formation.
The significance level at p<0.05 was
MATERIALS AND METHODS considered statistically significant. To correlate the
levels of calcium with the antioxidant activity,
The study included fifty diabetic subjects Pearson’s correlation co-efficient was worked out.
having cataract, aged between 45 -70 years of either
sex. Later they were grouped into group 1- immature
diabetic cataract patients and group 2- mature
diabetic cataract patients based on ophthalmoscopic
examination by ophthalmologist. Patients with
57 | International Journal of Life Sciences • ISSN 2091–0525 • Year 2012 • Volume 6 • Issue 1
Research Article Deepa et al. (2012)
N Immature N Mature
Dry weight of lens 24 56±1.79 26 70±2.57**
Total Proteins 57±0.56 65±0.66*
(mg/lens)
Antioxidant 1.320± 0.208 0.820± 0.085***
Activity
(mg/mg of protein)
Calcium 24 0.805± 0.416 26 1.024± 0.056***
(mg/mgof protein)
***=p<0.001
Figure 1. Comparison of lens antioxidant activity between Immature and Mature diabetic cataract patients
Figure 2. Comparison of lens calcium levels in Immature and Mature Diabetic Cataract patients
stress leads to increased intracellular calcium levels status which is responsible for opacification of lens
and in turn leads to cataract formation by following and also calcium plays an important role in causation
mechanisms, increased intracellular calcium interacts of diabetic cataract.
with cytoplasmic proteins causing protein oxidation
and aggregation of high molecular weight proteins. REFERENCES
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activity in human fluids. Journal of Clinical Pathology. 54:356- these), and authors who need assistance with language editing
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