Biological Evaluation of Polyherbal Ayurvedic Cardiotonic Preparation "Mahamrutyunjaya rasa"

Abstract

Mahamrutyunjaya rasa (MHR), an Ayurvedic formulation, used as cardiotonic, contains potentially toxic compounds like aconitine, which are detoxified during preparation using traditional methods. Comparative toxicological evaluation of laboratory prepared formulation (F1) and two marketed formulations (F2 and F3) were performed based on their effects on viability of H9c2 cells and after single oral dose administration in mice. Cardioprotective effect of formulations at 25 and 50 mg/kg doses were studied in isoproterenol- (ISO-) induced myocardial infarcted rats. F1 and F2 did not affect the cell viability, while F3 decreased the cell viability in concentration and time-dependent manner. Rats administered with ISO showed significant increase in the serum levels of glutamate oxaloacetate transaminase, alkaline phosphotase, creatinine kinase isoenzymes, lactate dehydrogenase, and uric acid, while F1 and F2 treatment showed significant reduction in the same. F3 showed further increase in the serum levels of enzymes and uric acid in ISO-challenged rats. High pressure liquid chromatographic analysis of formulations showed higher concentration of aconitine in F3. Study shows that F1 and F2 possess cardioprotective property with higher safety, while formulation F3 cannot be used as cardioprotective due to its cytotoxic effects. Thus, proper quality assessment methods are required during preparation of traditional formulations.

Figure 1

Microscopic images of mice heart, kidney, and liver illustrating effect of single oral dose administration of formulation F1, F2, and F3 (Staining: Hematoxylin and Eosin). Microscopic images of mice heart from (a) F1, (b) F2, (c) F3 groups; images of mice kidney from (d) F1, (e) F2, and (f) F3 images of mice liver from (g) F1, (h) F2, and (i) F3 groups. Solid arrow indicates the normal; and unfilled arrow indicates the affected/infracted/injured area of kidney/heart//liver.

Figure 2

Effect of formulations F1 (a), F2 (b), and F3 (c) on viability of H9c2 cells after 12, 24, and 48 h of treatment. Values are expressed as mean ± SD and are average of three determinations.

Figure 3

Effect of Formulations on serum levels of (a) CK-MB, (b) LDH, (c) GOT, (d) ALKP, and (e) Uric acid. Group I (1% solution of tween 80 in water followed by normal saline (2 mL/kg, s.c. twice at an interval of 24 h)); Group II (1% tween 80 followed by ISO (25 mg/kg, s.c. twice at an interval of 24 h)); Group III, VI, IX: (50 mg/kg dose of F1, F2, and F3, resp., followed by normal saline (2 mL/kg, s.c. twice at an interval of 24 h)); Group IV, VII, X: (administered orally with 25 mg/kg dose of F1, F2, and F3, respectively, followed by ISO (25 mg/kg, s.c. twice at an interval of 24 h)); Group V, VIII, XI (administered orally with 50 mg/kg dose of F1, F2, and F3, resp., followed by ISO (25 mg/kg, s.c. twice at an interval of 24 h)). #Compared with ISO treated group, *Compared with saline treated (control) group.

Figure 4

Effect of formulations F1, F2, and F3 (a) 25 mg/kg and (b) 50 mg/kg on Heart weight/body weight ratio (HW/BW). Group I (1% solution of tween 80 in water followed by normal saline (2 mL/kg, s.c. twice at an interval of 24 h)); Group II (1% tween 80 followed by ISO (25 mg/kg, s.c. twice at an interval of 24 h)); Group III, VI, IX: (50 mg/kg dose of F1, F2, and F3, resp., followed by normal saline (2 mL/kg, s.c. twice at an interval of 24 h)); Group IV, VII, X: (administered orally with 25 mg/kg dose of F1, F2, and F3, resp., followed by ISO (25 mg/kg, s.c. twice at an interval of 24 h)); Group V, VIII, XI (administered orally with 50 mg/kg dose of F1, F2, and F3, resp., followed by ISO (25 mg/kg, s.c. twice at an interval of 24 h)). *Compared with control group.

Figure 5

Microscopic images of rat heart illustrating the effect of formulation F1, F2, and F3 in ISO-induced MI rats (Staining: Haematoxylin and Eosin). Microscopic images of rat heart from Group I (a), Group II (b), Group III (c), Group IV (d), Group V (e), Group VI (f), Group VII (g), Group VIII (h), Group IX (i), Group X (j), Group XI (k).

Figure 6

Representative chromatogram of formulations F1 (prepared in laboratory), F2 (Baidyanath), and F3 (Pune Rasashala). Retention time of Aconitine ~8.2 mins. (Aconitine content (μgg⁻¹) in F1-1.06, F2-1.27, and F3-5.10.).

Figure 7

Schematic Representation of the study performed for the biological evaluation of Mahamrutyunjaya rasa.