INTERPLAY OF VITAMIN B6, L-TRYPTOPHAN AND NARINGENIN IN DOXORUBICIN INDUCED CORONARY ARTERY DAMAGE
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Date
2024-02
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SRI VENKATESWARA VETERINARY UNIVERSITY, TIRUPATI - 517 502. (A.P.) INDIA
Abstract
The doxorubicin induced damage in coronary artery was studied initially in
porcine coronary artery as in vitro model. The porcine heart was collected immediately
after slaughter in chilled Krebs-Henseleit solution (pH-7.4). The Right coronary artery
isolated was used for the in vitro experimentation. The RCA was placed in sterile petri
dishes in Krebs-Henseleit solution and the total of four treatments with six replications
was done. Group I served as control (Normal). Group II served as Doxorubicin control
where the RCA was incubated in 40 µM doxorubicin for 3 h. Group III, IV and Group V,
RCA were pre-incubated in 30 µM vitamin B6, tryptophan and naringenin for 1h
respectively and received 40 µM doxorubicin for 3 h.
After completion of treatment with doxorubicin, the tissue was utilized for
biochemical and histomorphological examinations. Biochemical tests viz., GSH, SOD,
TP and MDA were used to determine the status of oxidative damage. LDH and NO were
estimated to determine the extent of cytotoxicity. There is a fall in total protein levels and
antioxidant enzymes levels (GSH & SOD) in doxorubicin pre-treated group. Loss of
endothelium and inflammatory changes were evidenced in doxorubicin treated coronary
artery. There is raise in MDA, LDH and nitric oxide levels in doxorubicin pre-treated
group. Pre-treatment with vitamin B6, tryptophan and naringenin caused a protective
effect on doxorubicin treated coronary artery and restored the enzymes levels back to
normal. Regeneration of endothelium was observed in all three treatment groups upon
histomorphological examination.
In the in vivo experimentation with rabbits, 36 rabbits of 1.5-2 kg body weight
were randomly assigned to 6 groups of six rabbits in each group. Group I served as healthy
normal animals and Group II rabbits were given doxorubicin @10mg/kg B,Wt
intraperitoneally on 15th and 16th day of study. Group III animals received vitamin B6
@50 mg/kg B.Wt per orally for 14 days. Group IV and V received tryptophan (100 mg/kg
B.Wt) and naringenin (100 mg/kg B.Wt) per orally for 14 days. Group VI served as an
interplay group where the rabbits received vitamin B6 (50mg/kg B.Wt), tryptophan (100
mg/kg B.Wt) and naringenin (100 mg/kg B.Wt) for 14 days. On day 15 and 16, all groups
of rabbits received doxorubicin @10mg/kg B.Wt intraperitoneally.
On day 17 the rabbits were sacrificed using xylazine and ketamine anaesthesia
and heart samples were collected in chilled Krebs-Hanseleit solution. Half portion of the
heart tissues was used to prepare tissue homogenate to study antioxidant enzymes levels
and the remaining half to study histomorphological alterations. Blood was collected
during sacrifice to study heamatological parameters and the seperated serum was used to
estimate calcium, phosphorous, nitric oxide and lactate dehydrogenase. There is a fall in
levels of antioxidant enzymes (GSH and SOD), phosphorous, RBC, WBC, haemoglobin,
platelets count where as there is elevation of MDA levels, calcium, nitric oxide and LDH
levels in the doxorubicin treated rabbits. Gross morphological changes like pale heart,
congestion of blood vessels and fatty change and histomorphological alterations like loss
of endothelium, infiltration of inflammatory cells, vacuolation of cardiomyocytes were
observed in doxorubicin treated rabbits. Pre-treatment with vitamin B6, tryptophan,
naringenin and their combination restored the antioxidant enzymes levels, haematological
and cytotoxicity markers. Normal architecture of the artery was preserved in these rabbits
and gross changes were mild to moderate congestion. Rabbits of Group VI showed less
damage which may be due to synergistic effect of vitamin B6, tryptophan and naringenin
over doxorubicin damaged heart and coronary artery. Based on the above results, it is
concluded that combination of vitamin B6, tryptophan and naringenin offered better
protection on doxorubicin induced coronary artery damage.