Can the methanolic extract of Andrographis paniculata be used as a supplement to anti-snake venom to normalize hemostatic parameters: A thromboelastographic study

Abstract

Ethnopharmacological relevance of Andrographis paniculataThe whole plant (including leaves and roots) is used in traditional Ayurveda and Siddha medicine to treat various clinical conditions such as fever, respiratory tract infections, colic pain, liver disorders, diabetes, hypertension, and inflammation. It is also used as an antidote for snake-bite, poisonous bites of insects and recommended as a dietary supplement to boost immunity. Aim of the studyIn-vitro thromboelastographic evaluation of the efficacy of methanolic extract of Andrographis paniculata (MAP) and polyvalent anti-snake venom (ASV) in neutralizing the Naja naja (N.N) venom-induced changes in hemostatic parameters. Materials and methodsThromboelastographic evaluation of hemostatic parameters was initiated by adding 3μg N.N venom to citrated whole blood from healthy volunteers. The effect of different concentrations of ASV and MAP in neutralizing the toxicity of N.N venom were studied in two groups. In group 1 experiments, citrated whole blood (340μl) was mixed with N.N venom (3μg), immediately followed by successive addition of ASV (5μl, 8μl and 15μl) or MAP (15μg, 30μg, 60μg and 120 μg) or combination of ASV and MAP (3μl ASV+30μg MAP and 3μl ASV+60μg MAP). In group 2 experiments, N.N venom was incubated with whole blood for 90 minutes at 37°C, followed by successive addition of ASV (5μl, 10μl, and 15μl) or MAP (30μg and 60μg) or combination of ASV and MAP (5μl ASV+30μg MAP and 5μl ASV+60μg MAP). ResultIn Group 1 experiments, N.N venom caused significant (p<0.05) increase in R-time, K-time, LY30% and a decrease in angle and MA. Optimum effect on hemostatic parameters was observed at a concentration of 8μl ASV, where all the deleterious effects of the venom were completely reversed. Similarly, the addition of MAP to the assay system could reproduce results as ASV, in reversing the deleterious effects of the venom. This occurred in a concentration-dependent manner, from 15μg-60μg, with the optimum results at 60μg. When ASV concentration was reduced to 3μl and supplemented with MAP (30μg or 60μg), the positive supplementary effect of MAP was demonstrated.In Group 2 experiments, N.N venom caused significant (p<0.05) changes in all TEG parameters, with most deleterious changes observed in MA and LY30% compared to Group 1 experiments. ASV when added in increasing concentrations (5-15μl), had beneficial effects only on K-time, angle, and MA. When added together with ASV, MAP (30μg or 60μg) could significantly (p<0.05) supplement the effect of ASV (5μl) in improving R-time, K-time, and angle. ConclusionThis in-vitro study demonstrates the effectiveness of MAP as a supplement to ASV in combating the deleterious effects of N.N venom on hemostasis. However, further in-vivo experiments in animal models are required to substantiate these effects.