|Year : 2016 | Volume
| Issue : 2 | Page : 561-564
Protective effect of ayurvedic formulations against doxorubicin-induced cardiotoxicity: Preliminary studies on Brahma Rasayana and Chyavanaprash
Entissar AlSuhailbani1, Aditya Menon2, Cherupally Krishnan Krishnan Nair2
1 Department of Genetics, College of Science, King Saud University, Riyadh, Saudi Arabia
2 Pushpagiri Institute of Medical Sciences and Research Centre, Thiruvalla, Kerala, India
|Date of Web Publication||25-Jul-2016|
Cherupally Krishnan Krishnan Nair
Pushpagiri Institute of Medical Sciences and Research Centre, Thiruvalla - 689 101, Kerala
Source of Support: None, Conflict of Interest: None
Aim of Study: The present work aimed to examine the efficacy of two ayurvedic formulations, Brahma Rasayana (BRM) and Chyavanaprash (CHM) to alleviate doxorubicin (DOX) induced acute cardiotoxicity.
Materials and Methods: Swiss albino mice were administered with DOX (25 mg/kg, i.p.) and two doses of BRM or CHM (1 and 2 g/kg). Cardiotoxicity was assessed by measuring the levels of various antioxidant parameters in the heart as well as release of marker enzymes in the serum was assayed. Histology of the heart was also performed to check for DOX-induced damages.
Results: Administration of either BRM or CHM (1 and 2 g/kg) maintained the antioxidant status in the heart thereby preventing tissue damage as well as the release of marker enzymes. DOX-induced variation of cardiac architecture was also prevented by BRM and CHM administration.
Conclusion: BRM and CHM administration could prevent DOX-induced acute cardiotoxicity.
Keywords: Antioxidant status, Brahma Rasayana, cardiotoxicity, Chyavanaprash, doxorubicin
|How to cite this article:|
AlSuhailbani E, Menon A, Nair CK. Protective effect of ayurvedic formulations against doxorubicin-induced cardiotoxicity: Preliminary studies on Brahma Rasayana and Chyavanaprash. J Can Res Ther 2016;12:561-4
|How to cite this URL:|
AlSuhailbani E, Menon A, Nair CK. Protective effect of ayurvedic formulations against doxorubicin-induced cardiotoxicity: Preliminary studies on Brahma Rasayana and Chyavanaprash. J Can Res Ther [serial online] 2016 [cited 2019 Mar 22];12:561-4. Available from: http://www.cancerjournal.net/text.asp?2016/12/2/561/151931
| > Introduction|| |
Doxorubicin (DOX), an anthracycline cancer chemotherapeutic drug, is widely used to treat various types of malignancies. The long-term use of DOX is limited by its cardiotoxicity, which includes congestive heart failure, chronic cardiomyopathy, and changes in electrocardiogram pattern.
Endogenous antioxidant deficits have been suggested to play a major role in DOX-induced cardiomyopathy and heart failure. DOX elicit cardiotoxicity by various mechanisms such as by interfering with DNA repair, inducing DNA damage, free radicals generation, lipid peroxidation, mitochondrial damage, etc., and decreasing the activity of Na + K + ATPase. Since free radicals bring out the basic mechanism of DOX-induced cardiotoxicity, it is rational to consider antioxidants as therapeutics to mitigate its toxicity.
Ayurveda, the Indian holistic healthcare system of traditional medicines using natural ingredients, has several potent antioxidant formulations. Two of such formulations Brahma Rasayana (BRM) and Chyavanaprash (CHM) are evaluated in the present study for their cardioprotective ability since several ingredients of BRM and CHM are proved to be potent antioxidants., These formulations have shown promise in alleviating various free radicals induced physiological conditions., Present study envisages to find out the ability of these ayurvedic formulations in reducing DOX-induced cardiotoxicity in mice model.
| > Materials and Methods|| |
Brahma Rasayana and CHM, made by a renowned manufacturer were obtained from a retail outlet. All the other chemicals and reagents used in this study were of analytical grade.
Male swiss albino mice of 8–10 weeks old, weighing 22–25 g was obtained from the small animal breeding section, Kerala Agricultural University, Mannuthy, Thrissur, Kerala. They were kept under standard conditions of temperature and humidity in the center's animal house facility. The animals were provided with standard mouse chow (Sai Durga Feeds and Foods, Bengaluru, India) and water ad libitum. All animal experiments in this study were carried out with the prior approval of the Institutional Animal Ethics Committee and were conducted strictly adhering to the guidelines of committee for the purpose of control and supervision of experiments on animals constituted by the Animal Welfare Division of Government of India.
Animals were randomly divided into six groups of five each as detailed below. Group I was kept as the untreated control and all the other groups received DOX (25 mg/kg, i.p.). Group III and Group IV were administered with BRM or CHM (2 g/kg, p.o.) after 1 h of DOX injection, i.p.
Group I: Untreated control.
Group II: DOX (25 mg/kg, i.p.).
Group III: DOX (25 mg/kg, i.p.) + BRM (1 g/kg, p.o.).
Group IV: DOX (25 mg/kg, i.p.) + BRM (2 g/kg, p.o.).
Group V: DOX (25 mg/kg, i.p.) + CHM (1 g/kg, p.o.).
Group VI: DOX (25 mg/kg, i.p.) + CHM (2 g/kg, p.o.).
Assessment of cardiotoxicity
Twenty-four hours after DOX treatment, blood was collected by cardiac puncture and serum was separated for biochemical analysis. Heart was excised and washed with ice-cold phosphate-buffered saline (PBS) and homogenates 10% (w/v) were prepared in PBS. Assessment of cardiac biomarkers such as creatinine kinase isoenzyme (CK-MB), lactate dehydrogenase (LDH), and glutamate oxaloacetate transaminase (GOT), were analyzed using diagnostic kits (Agappe Diagnostic Pvt. Ltd.; Ernakulam, Kerala, India). Level of GSH was assayed by the method of Moron et al. (1979), based on the reaction with 5,5'-dithiobis-2-nitrobenzoic acid. Glutathione peroxidase (GPx) activity was measured based on the method of Hafeman et al., based on the degradation of H2O2. Activity of superoxide dismutase (SOD) was measured by nitroblue tetrazolium reduction method of McCord and Fridovich, protein levels in the tissue were measured by following the method of Lowry et al. Levels in peroxidation of membrane lipids were done based on the method of Buege and Aust (1978). For histopathological studies, heart was fixed in 10% formalin and embedded in paraffin wax. Sections of 5 micron thickness were made using a microtome and stained with hematoxylin-eosin.
The results are presented as mean ± standard deviation of the studied group. Statistical analyses of the results were performed using ANOVA with Tukey–Kramer multiple comparisons test.
| > Results|| |
[Table 1] gives the cellular antioxidant levels (GSH, GPx, and SOD). The results indicated that a single dose of DOX (25 mg/kg) significantly decreased the tissue antioxidants. Oral administration of BRM or CHM helped to maintain these antioxidants at near normal levels.
|Table 1: Effect of BRM or CHM onantioxidants level in DOX induced cardiotoxicity in mice|
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Peroxidation of membrane lipids
As presented in [Figure 1], DOX (25 mg/kg) injection resulted in an increase in the levels of peroxidation of membrane lipids from 1.69 ± 0.30 to 4.1 ± 0.44 nM/mg protein. Administration of BRM or CHM helped in reducing the extent of peroxidation. BRM administration at a dose of 1 g/kg and 2 g/kg restored the level of peroxidation to 2.77 ± 0.62 and 2.01 ± 0.61, respectively. CHM administration at a dose of 1 g/kg and 2 g/kg restored the level of peroxidation to 2.63 ± 0.14 and 2.35 ± 0.29, respectively.
|Figure 1: Effect of Brahma Rasayana or Chyavanaprash on peroxidation of membrane lipids in doxorubicin-induced cardiotoxicity in mice. Values are expressed as mean ± standard deviation (n = 5)|
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Serum biochemical parameters
The levels of various serum parameters [Table 2] suggest that DOX (25 mg/kg) administration increased these enzyme levels while the administration of BRM or CHM prevented the increase in the serum enzyme levels following DOXO treatment.
|Table 2: Effect of BRMor CHM on serum marker enzymes in doxorubicin induced cardiotoxicity in mice|
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Treatment of animals with DOXO imposed damage to the cardiac tissues as can be observed from the histopathological analysis [Figure 2]. [Figure 2]a is the section of a normal heart which shows normal morphology. There was a loss in the myofibril content, vacuolization of the cytoplasm along with swelling of mitochondria in DOX (25 mg/kg) administered animals [Figure 2]b. Administration of BRM or CHM prevented these side effects of DOX toxicity [Figure 2]c,[Figure 2]d,[Figure 2]e,[Figure 2]f. The results of histopathological analysis correlated with the findings from the tissue antioxidant levels and serum marker enzymes.
|Figure 2: Effect of Brahma Rasayana and Chyavanaprash various doses (1 and 2 g/kg) on doxorubicin (20 mg/kg) induced cardiotoxicity in swiss albino mice (×40)|
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| > Discussion|| |
The clinical use of DOX, an anthracycline derivative used for the treatment of various types of cancer, is limited by its dose-limiting cardiotoxicity. The present work was an attempt to find the ability of two ayurvedic formulations in mitigating DOX-induced cardiotoxicity.
Doxorubicin elicits its cytotoxic effect by the generation of free radicals, by mainly two pathways: A nonenzymatic pathway that utilizes iron and an enzymatic mechanism using the mitochondrial respiratory chain. Ultimately, DOX catalyzes the reduction of oxygen to form superoxide radical  thereby putting stress on the total tissue antioxidant capacity. Increase in production of free radicals along with the endogenous deficiency of antioxidants in heart results in induction of cardiac stress, which can be measured by the decrease in the levels of various antioxidants as well as an increase in peroxidation of membrane lipids in heart tissue. Administration of BRM or CHM prevented the DOX-induced cardiotoxicity by restoring the total antioxidant content.
In the present study, a single dose of DOX (25 mg/kg) induced cardiotoxicity manifested biochemically by the increase in the levels of serum LDH and CK-MB. These results were consistent with previous studies reported by other investigators.,, The increased levels of LDH and CK-MB in serum suggest leakage of these enzymes from heart mitochondria because of DOX exerted toxicity in the tissue. Serum GOT is another enzyme whose increase has also been monitored during cardiac injury. Administration of BRM and CHM prevented the release of these marker enzymes from mitochondria by protecting it from free radical-induced damages of DOX.
The present study convincingly proved that BRM or CHM administration prevents the cardiotoxic effect of DOX. Screening the vast repertoire of ayurveda can yield in the discovery of formulations which can be useful in preventing the side effects of chemotherapeutic drugs. Further studies will throw more light whether these formulations can offer preferential protection to normal tissues without reducing the curative effect of the chemotherapeutics.
| > Acknowledgement|| |
The authors express their sincere appreciation to the Deanship of Scientific Research at King Saud University for support extended to the research group No (IRG14-05) for the publication
| > References|| |
Wang JJ, Cortes E, Sinks LF, Holland JF. Therapeutic effect and toxicity of adriamycin in patients with neoplastic disease. Cancer 1971;28:837-43.
Chlebowski RT. Adriamycin (doxorubicin) cardiotoxicity: A review. West J Med 1979;131:364-8.
Bristow MR, Mason JW, Billingham ME, Daniels JR. Doxorubicin cardiomyopathy: Evaluation by phonocardiography, endomyocardial biopsy, and cardiac catheterization. Ann Intern Med 1978;88:168-75.
Arola OJ, Saraste A, Pulkki K, Kallajoki M, Parvinen M, Voipio-Pulkki LM. Acute doxorubicin cardiotoxicity involves cardiomyocyte apoptosis. Cancer Res 2000;60:1789-92.
Lenaz L, Page JA. Cardiotoxicity of adriamycin and related anthracyclines. Cancer Treat Rev 1976;3:111-20.
Hanaa A, Fathia M, Gamal AE, Senot HD. Cardioprotective role of melatonin and its novel synthesized derivatives on doxorubicin induced cardiotoxicity. Bioorg Med Chem 2005;13:1847-57.
Dorr RT. Cytoprotective agents for anthracyclines. Semin Oncol 1996;23:23-34.
Myers CE, McGuire WP, Liss RH, Ifrim I, Grotzinger K, Young RC. Adriamycin: The role of lipid peroxidation in cardiac toxicity and tumor response. Science 1977;197:165-7.
Bier CC, Jaenke RS. Function of myocardial mitochondria in the adriamycin-induced cardiomyopathy of rabbits. J Natl Cancer Inst 1976;57:1091-4.
Geetha A, Devi CS. Effect of doxorubicin on heart mitochondrial enzymes in rats: A protective role for alpha-tocopherol. Indian J Exp Biol 1992;30:615-8.
Patwardhan B, Vaidya AD, Chorghade M, Joshi SP. Reverse pharmacology and systems approaches for drug discovery and development. Curr Bioact Compd 2008;4:201-12.
Ramnath V, Rekha PS, Sujatha KS. Amelioration of heat stress induced disturbances of antioxidant defense system in chicken by brahma rasayana. Evid Based Complement Alternat Med 2008;5:77-84.
Parle M, Bansal N. Traditional medicinal formulation, Chyawanprash – A review. Indian J Tradit Knowledge 2006;5:484-8.
Jose JK, Kuttan R. Hepatoprotective activity of Emblica officinalis
and Chyavanaprash. J Ethnopharmacol 2000;72:135-40.
Rekha PS, Kuttan G, Kuttan R. Antioxidant activity of brahma rasayana. Indian J Exp Biol 2001;39:447-52.
Moron MS, Depierre JW, Mannervik B. Levels of glutathione, glutathione reductase and glutathione S-transferase activities in rat lung and liver. Biochim Biophys Acta 1979;582:67-78.
Hafeman DG, Sunde RA, Hoekstra WG. Effect of dietary selenium on erythrocyte and liver glutathione peroxidase in the rat. J Nutr 1974;104:580-7.
McCord JM, Fridovich I. Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein). J Biol Chem 1969;244:6049-55.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193:265-75.
Buege JA, Aust SD. Microsomal lipid peroxidation. Methods Enzymol 1978;52:302-10.
Galeano M, Altavilla D, Cucinotta D, Russo GT, Calò M, Bitto A, et al.
Recombinant human erythropoietin stimulates angiogenesis and wound healing in the genetically diabetic mouse. Diabetes 2004;53:2509-17.
Booser DJ, Hortobagyi GN. Anthracycline antibiotics in cancer therapy. Focus on drug resistance. Drugs 1994;47:223-58.
Horenstein MS, Vander Heide RS, L'Ecuyer TJ. Molecular basis of anthracycline-induced cardiotoxicity and its prevention. Mol Genet Metab 2000;71:436-44.
Mimnaugh EG, Trush MA, Bhatnagar M, Gram TE. Enhancement of reactive oxygen-dependent mitochondrial membrane lipid peroxidation by the anticancer drug adriamycin. Biochem Pharmacol 1985;34:847-56.
Pacher P, Liaudet L, Bai P, Virag L, Mabley JG, Haskó G, et al.
Activation of poly (ADP-ribose) polymerase contributes to development of doxorubicin-induced heart failure. J Pharmacol Exp Ther 2002;300:862-7.
Dowd NP, Scully M, Adderley SR, Cunningham AJ, Fitzgerald DJ. Inhibition of cyclooxygenase-2 aggravates doxorubicin-mediated cardiac injury in vivo
. J Clin Invest 2001;108:585-90.
Saad SY, Najjar TA, Al-Rikabi AC. The preventive role of deferoxamine against acute doxorubicin-induced cardiac, renal and hepatic toxicity in rats. Pharmacol Res 2001;43:211-8.
Jagetia GC, Reddy TK, Malagi KJ, Nayak BS, Naidu MB, Ravikiran PB, et al.
Antarth, a polyherbal preparation protects against the doxorubicin-induced toxicity without compromising its Antineoplastic activity. Phytother Res 2005;19:772-8.
[Figure 1], [Figure 2]
[Table 1], [Table 2]