Investigating the effect of Withania somnifera on coagulation, blood count and bronchial lavage of Balb/c mice

Abstract

Withania somnifera is a phytomedicine that is an important component of the Ayurvedic Pharmacopoeia of India and has been used for over 400 years to treat all kinds of diseases and ailments. The aim of this study was to investigate the effect of W. somnifera on coagulation, blood count and bronchial lavage by using the asthmatic Balb/c mouse model. Because it’s known that asthmatic patients have decreased selenium levels and since selenium is known to be a potent antioxidant, the effects of selenium alone and in combination with Withania somnifera were investigated in this study. Hydrocortisone is most commonly used in the treatment of asthma and therefore hydrocortisone was used as positive control in this study. To establish the asthmatic Balb/c mouse model, the mice were sensitized, nebulized and treated over a period of 43 days and blood smears were made of each individual animal and bronchial lavage was performed by injecting 0.3ml of saline into the trachea of the mice on the day of termination. Light microscopy analysis of the bronchial lavage revealed a significant decrease in the number of eosinophils counted in the asthmatic and the different treatment groups. In the asthmatic group, numerous platelet clumps were found distributed between white blood cells. Platelets were also found in the other treatment groups but are not as prevalent as in the asthmatic group. Results from the blood smears showed the same trends, where cell counts in control and hydrocortisone blood smears were decreased compared to that of the asthma group. Platelets and fibrin play an important physiological role in allergic reactions and immunological mechanisms. In conditions like allergic asthma, platelets participate by acting as inflammatory cells, by releasing mediators, spasmogens and/or by interacting with other inflammatory cell types. Results indicate control mice possess major, thick fibers and minor thin fibers as well as tight round platelet aggregates with typical pseudopodia formation. Minor fibers of asthmatic mice have a netlike appearance covering the major fibers, while the platelets seem to form loosely connected, granular aggregates. Hydrocortisone made the fibrin more fragile and the platelet morphology changes form a tight platelet aggregate to a more granular aggregate not closely fused to each other. The plant extract separately and in combination with selenium did not impact on the fragility of the fibrin and reversed the formation of the dense minor netlike layer over the major fibers. A tight platelet aggregate was also seen in these mice. Asthmatic mice only treated with selenium showed a dense minor fibrin layer (as was seen in the asthmatic mice); however the platelets formed a dense aggregate. It is concluded that W. somnifera and W. somnifera combined with selenium significantly decreased the white blood cells in both bronchial lavage as well as blood smears, suggesting that W. somnifera indeed has an anti-inflammatory potential and it, in combination with an anti-oxidant like selenium might successfully be used in the treatment of asthma. Also, the anti-inflammatory products impacts on the fibrin network but not on the platelet aggregate stability (seen with hydrocortisone). Anti-oxidants like selenium do not impact on the stability of the fibrin networks in asthma, but impacts on platelet stability. Herbal products like W. somnifera, which are potent antioxidants as well as anti-inflammatory product impacts on the stability of both the fibrin networks and the platelet aggregates. These results therefore suggests that asthmatic patients should indeed use an antioxidant supplement e.g. selenium, as it stabilizes platelet aggregates.