Effect of Zinc Supplementation on Adenosine Deaminase ... - medIND
Effect of Zinc Supplementation on Adenosine Deaminase ... - medIND
Effect of Zinc Supplementation on Adenosine Deaminase ... - medIND
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376 Indian Medical Gazette — OCTOBER 2013<br />
Clinical Evaluati<strong>on</strong><br />
<str<strong>on</strong>g>Effect</str<strong>on</strong>g> <str<strong>on</strong>g>of</str<strong>on</strong>g> <str<strong>on</strong>g>Zinc</str<strong>on</strong>g> <str<strong>on</strong>g>Supplementati<strong>on</strong></str<strong>on</strong>g> <strong>on</strong><br />
<strong>Adenosine</strong> <strong>Deaminase</strong> Activity in Leprosy<br />
T. Pradhan, Dept <str<strong>on</strong>g>of</str<strong>on</strong>g> Biochemistry, Associate Pr<str<strong>on</strong>g>of</str<strong>on</strong>g>, KIMS, Bhubaneswar.<br />
S. Kumari, Dept <str<strong>on</strong>g>of</str<strong>on</strong>g> Biochemistry, Assistant Pr<str<strong>on</strong>g>of</str<strong>on</strong>g>, AIIMS, Bhubaneswar.<br />
Absract<br />
Serum <strong>Adenosine</strong> <strong>Deaminase</strong> (ADA) acts as marker<br />
<str<strong>on</strong>g>of</str<strong>on</strong>g> cellular immunity and its activity is found to be altered<br />
in various diseases in which there is a cell mediated<br />
immune resp<strong>on</strong>se (CMI) including leprosy. The role <str<strong>on</strong>g>of</str<strong>on</strong>g><br />
zinc is well established in the development and<br />
maintainence <str<strong>on</strong>g>of</str<strong>on</strong>g> immunocompetence and its supplementati<strong>on</strong><br />
activates the immune resp<strong>on</strong>se in particular T-<br />
lymphocytes and m<strong>on</strong>ocytes in several ways. The aim <str<strong>on</strong>g>of</str<strong>on</strong>g><br />
the study was planned to evaluate the effect <str<strong>on</strong>g>of</str<strong>on</strong>g> nutriti<strong>on</strong>al<br />
zinc supplementati<strong>on</strong> <strong>on</strong> cell mediated immune resp<strong>on</strong>se<br />
by investigating the pre and post interventi<strong>on</strong> serum ADA<br />
levels after oral zinc sulphate supplementati<strong>on</strong> in leprosy<br />
patients.<br />
A total <str<strong>on</strong>g>of</str<strong>on</strong>g> 49 cases, 30 Tuberculoid Leprosy (TT) and<br />
19 Lepromatous Leprosy (LL) patients, within the age<br />
group <str<strong>on</strong>g>of</str<strong>on</strong>g> 25-60 years were enrolled in the study al<strong>on</strong>g with<br />
30 age matched healthy c<strong>on</strong>trols. Serum ADA was<br />
estimated in all the subjects before and after (2 m<strong>on</strong>ths and<br />
4 m<strong>on</strong>ths) oral zinc supplementati<strong>on</strong>. Pre interventi<strong>on</strong> serum<br />
ADA level was observed to be significantly increased in<br />
both the TT and LL (p
Indian Medical Gazette — OCTOBER 2013 377<br />
functi<strong>on</strong>s <str<strong>on</strong>g>of</str<strong>on</strong>g> neutrophils, natural killer cells and<br />
macrophages.<br />
Keeping in view the above facts, this study was designed<br />
to find out the pre and post interventi<strong>on</strong> serum ADA level<br />
after oral zinc sulphate supplementati<strong>on</strong> and made an attempt<br />
to explore the beneficial effects <str<strong>on</strong>g>of</str<strong>on</strong>g> zinc and its potential as<br />
an nutriti<strong>on</strong>al immunostimulant in leprosy cases.<br />
Materials and Methods<br />
Forty nine (49) untreated patients visiting the OPD <str<strong>on</strong>g>of</str<strong>on</strong>g><br />
the leprosy home at Naya Bazaar, Cuttack, with definitive<br />
evidence <str<strong>on</strong>g>of</str<strong>on</strong>g> hypopigmented anaesthetic patches and<br />
enlarged peripheral nerves were included in this study. They<br />
were categorized according to the Ridley Jopling<br />
classificati<strong>on</strong> 16 into thirty (30) Tuberculoid Leprosy (TT)<br />
and nineteen (19) Lepromatous Leprosy (LL) cases. Slit<br />
skin smear and skin biopsy was carried out in all patients<br />
to c<strong>on</strong>firm the diagnosis. Thirty (30) age, sex and socio<br />
ec<strong>on</strong>omic status matched healthy pers<strong>on</strong>nel working at the<br />
leprosy home, not <strong>on</strong> any type <str<strong>on</strong>g>of</str<strong>on</strong>g> medicati<strong>on</strong> were taken as<br />
c<strong>on</strong>trols. Seum was estimated in all the leprosy cases and<br />
c<strong>on</strong>trols. After obtaining informed c<strong>on</strong>sent, the patients<br />
received zinc sulphate formulati<strong>on</strong> (220 mg/ day) for 4<br />
m<strong>on</strong>ths. During the treatment period they were advised<br />
not to take various drugs or any food products al<strong>on</strong>g with<br />
the supplement that would interfere with zinc absorpti<strong>on</strong>.<br />
Patients with history <str<strong>on</strong>g>of</str<strong>on</strong>g> Multi-Drug Therapy, cases with<br />
chr<strong>on</strong>ic c<strong>on</strong>diti<strong>on</strong>s such as leukemia, lymphoma, hepatitis,<br />
cirrhosis or any other chr<strong>on</strong>ic illness were excluded from<br />
the study.<br />
The present study was c<strong>on</strong>ducted in the department <str<strong>on</strong>g>of</str<strong>on</strong>g><br />
Biochemistry, Hi-Tech Medical College, Bhubaneswar,<br />
Orissa. Both cases and c<strong>on</strong>trols were subjected to routine<br />
biochemical investigati<strong>on</strong>s. Serum ADA activity was<br />
estimated by the method <str<strong>on</strong>g>of</str<strong>on</strong>g> Guisti and Galanti 11 . Amm<strong>on</strong>ia<br />
liberated by reacti<strong>on</strong> <str<strong>on</strong>g>of</str<strong>on</strong>g> ADA <strong>on</strong> adenosine forms an intensely<br />
blue indophenol with sodium hypochlorite and phenol in<br />
alkaline soluti<strong>on</strong>. The intensity <str<strong>on</strong>g>of</str<strong>on</strong>g> colour formed was<br />
measured spectrophotometrically at 620nm. One unit <str<strong>on</strong>g>of</str<strong>on</strong>g><br />
ADA is defined as the amount <str<strong>on</strong>g>of</str<strong>on</strong>g> enzyme required to release<br />
1 mmol <str<strong>on</strong>g>of</str<strong>on</strong>g> amm<strong>on</strong>ia/min from adenosine at standard assay<br />
c<strong>on</strong>diti<strong>on</strong>s. Results were expressed as internati<strong>on</strong>al unit <str<strong>on</strong>g>of</str<strong>on</strong>g><br />
enzyme activity <str<strong>on</strong>g>of</str<strong>on</strong>g> serum. Estimati<strong>on</strong> was d<strong>on</strong>e in the preinterventi<strong>on</strong><br />
period, and after 2 m<strong>on</strong>ths and 4 m<strong>on</strong>ths oral<br />
zinc therapy after which results were tabulated after<br />
statistical analysis. There were no drop-outs during 2<br />
m<strong>on</strong>ths supplementati<strong>on</strong> period, however, after 4 m<strong>on</strong>ths,<br />
ADA measurement could be d<strong>on</strong>e in 38 cases as 4 TT and<br />
7 LL subjects withdrew their c<strong>on</strong>sent, either because <str<strong>on</strong>g>of</str<strong>on</strong>g> GI<br />
side effects or due to lack <str<strong>on</strong>g>of</str<strong>on</strong>g> motivati<strong>on</strong>. Results were<br />
expressed in Mean ± SD. Statistically analysis <str<strong>on</strong>g>of</str<strong>on</strong>g> results<br />
was d<strong>on</strong>e using student’s ‘t’ test.<br />
Observati<strong>on</strong>s<br />
As the cases bel<strong>on</strong>ged to a low socio-ec<strong>on</strong>omic status,<br />
routine biochemical parameters like Hb% FBS and Total<br />
serum protein revealed a moderate fall (p< 0.05) indicating<br />
a state <str<strong>on</strong>g>of</str<strong>on</strong>g> hypoproteinemia and anaemia while TLC and<br />
ESR showed a rise (p
.<br />
378 Indian Medical Gazette — OCTOBER 2013<br />
Table 2<br />
Baseline ADA (IU/L) Levels in the Study Group<br />
C<strong>on</strong>trols ( n=30) TT (n=30) LL (n=19)<br />
20 ± 2.31 47.04 ± 5.37* 30.23 ± 7.52*<br />
*( p< 0.001), highly significant<br />
immune resp<strong>on</strong>se against the invading mycobacteria.<br />
However raised ADA activity seen in an immune<br />
hyporresp<strong>on</strong>sive state like LL may be due to abnormal<br />
proliferati<strong>on</strong> <str<strong>on</strong>g>of</str<strong>on</strong>g> different subsets <str<strong>on</strong>g>of</str<strong>on</strong>g> lymphocytes. Table 3<br />
showed variable changes after zinc sulphate supplementati<strong>on</strong><br />
in the diseased groups. After 2 m<strong>on</strong>ths, there was no<br />
significant change (NS) and a moderate rise (p
Indian Medical Gazette — OCTOBER 2013 379<br />
activity in LL cases by the effect <str<strong>on</strong>g>of</str<strong>on</strong>g> oral zinc therapy<br />
indicating an improvement in several immune parameters 6 .<br />
Some experimental studies have c<strong>on</strong>cluded that it is possible<br />
to restore the enzyme activity in aged skin fibroblasts to<br />
normal levels by raising the zinc c<strong>on</strong>centrati<strong>on</strong> in the culture<br />
medium to four or five times the c<strong>on</strong>trol normal plasma<br />
zinc level 10 while others suggested that though ZnCl 2<br />
has<br />
enhancement effects <strong>on</strong> ADA activity in a dose dependent<br />
manner, it might show toxicity with deleterious effects in<br />
higher c<strong>on</strong>centrati<strong>on</strong>s 21 . The above results matched with<br />
the present study which depicts that oral zinc therapy plays<br />
an important role in specific immune defences and<br />
microbicidal activity in both types <str<strong>on</strong>g>of</str<strong>on</strong>g> leprosy more so in<br />
the TT than LL group by significantly enhancing the ADA<br />
activity in the cases.<br />
Summary<br />
ADA as a marker <str<strong>on</strong>g>of</str<strong>on</strong>g> CMI, c<strong>on</strong>sistent with lymphocyte<br />
and macrophage activity was found to be raised in leprosy<br />
patients in comparis<strong>on</strong> to c<strong>on</strong>trols. Its c<strong>on</strong>centrati<strong>on</strong> runs<br />
parallel with the immunological resp<strong>on</strong>se <str<strong>on</strong>g>of</str<strong>on</strong>g> the patients<br />
against antigenic stimulati<strong>on</strong>. Administrati<strong>on</strong> <str<strong>on</strong>g>of</str<strong>on</strong>g> zinc brings<br />
about stimulati<strong>on</strong> <str<strong>on</strong>g>of</str<strong>on</strong>g> a variety <str<strong>on</strong>g>of</str<strong>on</strong>g> immunological functi<strong>on</strong>s<br />
involved in host defense like T-lymphocyte and macrophage<br />
activati<strong>on</strong> indicated by a rise in serum ADA levels after<br />
supplementati<strong>on</strong> in leprosy patients. <str<strong>on</strong>g>Zinc</str<strong>on</strong>g> having the ability<br />
to enhance cell mediated immunity, could reduce the<br />
incidence <str<strong>on</strong>g>of</str<strong>on</strong>g> reacti<strong>on</strong> and minimize the morbidity <str<strong>on</strong>g>of</str<strong>on</strong>g> the<br />
disease. In additi<strong>on</strong> to various drugs and other primary<br />
immunomodulators, it has an immense immunotherapeutic<br />
role in treating leprosy cases and maybe used as an adjunct<br />
to chemotherapy to enhance bacterial killing as well as<br />
bacterial clearance and hence shorten the treatment period.<br />
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