Effect of Nardostachys jatamansi DC. on Apoptosis, Inflammation and Oxidative Stress Induced by Doxorubicin in Wistar Rats

Abstract

The study aimed to investigate the protective action of jatamansi (Nardostachys jatamansi DC.) against doxorubicin cardiotoxicity. Methanolic extract of jatamansi (MEJ) was prepared and standardized using HPTLC fingerprinting, GC-MS chemoprofiling, total phenolic content, and antioxidant activity in vitro. Further in vivo activity was evaluated using rodent model. Animals were divided into five groups (n = 6) namely control (CNT) (Normal saline), toxicant (TOX, without any treatment), MEJ at low dose (JAT1), MEJ at high dose (JAT2), and standard desferrioxamine (STD). All groups except control received doxorubicin 2.5 mg per Kg intra-peritoneally for 3 weeks in twice a week regimen. After 3 weeks, the blood samples and cardiac tissues were collected from all groups for biochemical and histopathological evaluation. Treatment with MEJ at both dose levels exhibited significant reduction (p < 0.001 vs. toxicant) of serum CK-MB (heart creatine kinase), LDH (Lactate dehydrogenase) & HMG-CoA (3-hydroxy-3-methylglutaryl-coenzyme A) levels, and tissue MDA (melondialdehyde) level; insignificant difference was observed (p > 0.05) in TNF-alpha (tumour necrosis factor), IL-6 (interleukine-6) levels and caspase activity as compared to TOX. Histopathological evaluation of cardiac tissues of different treatment groups further reinforced the findings of biochemical estimation. This study concludes that jatamansi can protect cardiac tissues from oxidative stress-induced cell injury and lipid peroxidation as well as against inflammatory and apoptotic effects on cardiac tissues.

Keywords: Nardostachys jatamansi DC.; biochemical estimation; cardioprotection; cardiotoxicity; doxorubicin; lipid peroxidation.

Figure A1

GC-MS chromatogram of hexane fraction of MEJ showing jatamansone as main constituent.

Figure A2

Mass spectra of jatamansone in sample (A) comparing with library (B) by GC-MS.

Figure 1

Nardostachys jatamansi.

Figure 2

Antioxidant potential of MEJ compared with ascorbic acid by using DPPH (A) and Nitric oxide (B) scavenging method.

Figure 3

Results of different biochemical parameters for CNT group, TOX group, STD, MEJ 250 mg/kg (JAT1), and 500 mg/kg treated group (JAT2). (A) serum CK-MB level where TOX group showed a significant increase (p < 0.01 vs. Control) in CK-MB activity, indicating myocardial damage. Treatment with Jatamansi exhibited a significant reduction in CK-MB activity. (B) serum LDH activity was significantly increased with DOX treatment. This effect was reversed upon treatment with Jatamansi. (C)Tissue MDA (TBARS) level was elevated significantly compared to control group, which indicates severe lipid peroxidation and oxidative stress. Jatamansi at both dose levels prevented elevation of MDA levels significantly; (D) serum HMG-CoA level, which was similar to other mentioned parameters, was significantly low in Jatamansi-treated animals; Serum inflammatory marker TNF-α (E) and IL-6 (F) also showed elevated levels upon DOX treatment. The anti-inflammatory effect of Jatamansi was evident from lowering of TNF- α and IL-6 levels as compared to DOX. (G) % caspase activity measured as marker of apoptosis showed a significant increase in cardiac tissue of DOX-treated animals.

Figure 4

Histopathology of cardiac tissues from animals belonging to CNT, TOX group, MEJ groups, and STD-treated group was assessed with special reference to the integrity of myocardial fibre integrity and evidence of cardiac damage caused by DOX. CNT shows normal myocardial structure, whereas TOX shows disarray of myocardial cells with small and large vacuolar myopathy but no evidence of necrosis of the myocardium seen. Cardiac tissue from MEJ (JAT1, JAT 2) and STD groups, respectively, shows normal myocardial architecture.