GC-MS analysis
The GC-MS analysis of the ethanolic extract of
P. pterocarpum leaves is displayed in Fig 1. Eight peaks were detected over the retention period, which spanned from 11.30 to 44.24 minutes. The compounds are psoralene,cyclohexasiloxane-dodecamethyl, hexadecanoic acid, cycloheptasiloxane-tetradecamethyl, 4-ethyl-8-quinolinol, docosanoic acid, canthaxanthin, vitamin E acetate.
Effect of P. pterocarpum leaves on lipid profiles in doxorubicin induced rats
The results of the serum lipid profile levels in control and experimental groups were shown in the in Table 1. The level of lipid profiles except HDL, were increased in doxorubicin induced rats. The levels of the all parameters were normalized nearer to the control when the treatment of ethanolic extract of
P. pterocarpum leaves and standard drug propranolol treated groups.
The increasing concentration of cholesterol in heart tissues is induces the oxidative stress and it induces the formation of free radicals and it leads the cellular injury (
Gokkusu and Mostafa Zadeh, 2003).
Anandan et al. (2007) demonstrated the effect of doxorubicin, which raises the cholesterol level in the rats. This proved in the present study also. After the treatment of
P. pterocarpum leaves the levels returned to normal. This could be because the phytocompounds slow down the production of free radicals and shield the cells from injury.
The increasing level of cholesterol may induce the synthesis of fatty acids and TG levels in the liver and heart tissues. The increasing concentration of TG is directly proportional to the cardiovascular disease
(Fungwe et al., 1993). In the present study, the level of TG is elevated in the doxorubicin treated rats. After the treatment of
P.
pterocarpum leaves the level of TG was normalized. This might be due to the cholesterol reduction because the reducing level of cholesterol may reduces the fatty acid synthesis in serum. The previous study of
Subashini et al., (2007) proves that the Nardostachys jatamansi extract reduces the TG level in the doxorubicin induced cardio toxicity rats.
The increased concentration of LDL is directly proportional to the risk of myocardial infarction. At the same time, the decreased level of TG may reduced the level of LDL and VLDL. In the present study, the level of LDL and VLDL was increased in the doxorubicin treated rats. After the treatment the level of LDL and VLDL was normalized. This demonstrates clearly that the plant extract lowered the risk of atherosclerosis. The VLDL level is directly correlated with the declining TG concentration.
Sakthivel et al. (2010) and
Manikandan et al. (2018) found comparable results to these. This study showed that when doxorubicin is used to treat cardiotoxicity-induced albino rats, the levels of LDL and VLDL in
Buchanania axillaris are lowered. The previous study of
Algefare et al., (2022) also proves the same.
The transfer of cholesterol from the tissues to the liver is greatly aided by this HDL. This is because lipoprotein lipase activity lowers HDL levels. In the current investigation, rats given doxorubicin had lower levels of HDL. The amount of HDL increased following treatment with the ethanolic extract of
P. pterocarpum leaves. The level of HDL is indirectly proportional to the risk of cardiac damage.
Effect of P. pterocarpum leaves extracts on the lysosomal enzymes
In the doxorubicin induced rats the level of lysosomal enzymes were increased in the experimental rats when compared to the control rats. The treatment of ethanolic extract of
P. pterocarpum leaves reduced the levels significantly (Table 2).
Myocardial cellular damage is caused by the release of the enzyme acid phosphatase from the lysosome into the cytosol by the acid hydrolases in the heart (Decker and Wildentha, 1980). In the present study, after the treatment of
P. pterocarpum leaves the levels were reduced when compared to the disease control rats. This might be caused by lysosomal hydrolases’ decreased activity. This work may corroborate the earlier research by Karthikeyan
et al. (2007), which demonstrated that treatment with grape seed proanthocyanidins lowers serum levels of acid phosphatase.
In the current investigation, the level of cathepsin D was significantly decreased after the treatment of
P. pterocarpum leaves in the cardiotoxicity rats. This might be as a result of the leaf extracts potent inhibition of lysosomal enzyme release, which may shield membranes from damage and boost lysosome stability.
Arafa et al., (2014) proved the cardioprotective properties of endangered Indian medicinal plants. Researchers demonstrated that ethanolic extracts of
Gmelina arborea and
Grewiaum bellifera considerably lower the activity of cathepsin D in rats given doxorubicin.
In the current investigation, the levels of β-D-glucuronidase and β-N-acetyl glucosaminidase levels were significantly reduced after the treatment of
P. pterocarpum leaves in the cardiotoxicity rats. This could be attributed to plants anti-oxidant activity, as it scavenge for oxygen free radicals, which may have conserved the stability of lysosomes and cell membranes. This could stop the lysosomal enzymes from leaking out.
Suchalatha and Devi (2004) demonstrated that
T. chebula treatments in rats with isoproterenol-induced heart injury have lower levels of β-D-glucuronidase and β-N-acetyl glucosaminidase.
Effect of P. pterocarpum leaves extracts on the mitochondrial enzymes
The level of all enzymes (ICH, KDH, SDH, MDH, NADPH dehydrogenase and cytochrome C oxidase) were reduced in the doxorubicin induced rats (Table 3). The treatment of ethanolic extract of
P. pterocarpum leaves the levels were returned back to the normal compared to the control rats.
The level of ICH was reduced due to the damage of the mitochondrial by the action of doxorubicin. Because of the stress caused by the rising amounts of free radicals produced by the doxorubin-induced rats, the levels of tricarboxylic acid cycle enzymes are decreased. The current findings were synchronized with the
Ganoderma lucidum versus CCl4 treated rats results (
Sudheesh, 2011).
The tricarboxylic acid cycle enzymes activities are increased in the current study by the ethanolic leaf extract of
P. pterocarpum and propranolol. This could strengthen the mitochondrial anti-oxidant defense system and alleviate the problems associated with the tricarboxylic acid cycle’s lower function. Propranolol and
P. pterocarpum leaf extract treatment may lessen oxidative damage to mitochondria and improve anti-oxidant status. Al-Assaf (2014) suggested that the decrease in mitochondrial anti-oxidants may be attributed to either feed-back inhibition or oxidative inactivation of enzyme proteins resulting from excessive ROS production.
Effect of P. pterocarpum leaves extracts on the cardiac marker enzymes
The levels of CK-MB, CK and troponin levels were increased in the doxorubicin induced rats when compared to the normal rats. After the treatment of ethanolic extract of
P.
pterocarpum leaves the levels were return back to the normal when compared to the normal and standard drug treated rats (Table 4)
.
In animal models, myocardial infarction is directly correlated with rising blood troponin levels
(O’Brien et al., 2006). Myofibrillarlysis, which results in the release of troponin, can be caused by decreased production of muscle-specific proteins, accelerated breakdown of myofibrillar components. and suppression of protein synthesis. The level of troponin was elevated in the doxorubicin-induced rats; this could be because to tissue injury and inhibition of protein synthesis. The extract which decreased the protein breakdown and it favors the protein formation. This effect is reflected in the decreased release of troponin into the bloodstream from cardiac muscle;
Herman et al. (1998) had shown the value of troponin as a marker for detecting cardio toxicity; significant (P<0.05) differences were noted between the treatment groups.
After the treatment of
P. pterocarpum leaves the levels of CK was decreased. The toxins are directly attacks the plasma membrane permeability. The leaves of
P. pterocarpum may reduce
s the damage and increases the plasma membrane permeability and this may reduces the CK secretion in the blood stream. Both the usual medication treated rats and the rats treated with the plant extract alone had levels that were comparable to those of the control group.
In the time of myocardial damage, the CK-MB enzyme is released from the heart into the circulatory blood. The toxins rupture the cells and releases the CK-MB from heart into the blood. In the present study, after the treatment of
P. pterocarpum leaves the levels are decreased. Significant (P<0.05) changes were noted in the treatment groups.
Grewia umbellifera and
Gmelina arborea ethanolic extracts reduce CK-MB activity in rats treated with doxorubicin, as demonstrated by
Arafa et al. (2014).
In silico analysis of the compound canthaxanthin reported from P. pterocarpum leaves
In the present study, the
in silico approach on phytochemicals canthaxanthin against cardiac target of creatine kinase-Muscle, Cretine kinase-Brain and troponin is carried out using virtual screening, molecular docking and ADMET methods. Virtual screening of canthaxanthin compound showed the binding affinity towards target creatine kinase-Muscle, Cretine kinase-Brain and troponin.
The docking scores were canthaxanthin was found by-6.331, -5.289 and -6.369 Kcal/mol to the creatine kinase-brain, creatine kinase-muscle and troponin resptctively. (Table 5, Fig 2-4). The molecular docking of the hits showed the binding mode and interaction energy. The docking studies confirmed the inhibition of cardiac target protein creatine kinase-brain, creatine kinase-muscle and troponin to show the cardioprotctive activity of canthaxanthin.
