Effects Of Centella Asiatica Extracts On Cell Cycle And Intracellular "Ca²⁺i Of Spleen Lymphocytes

ObjectiveTo isolate the Further purification of antioxidant activite ingredient from centella asiatica and evaluate its antioxidant potential; Lymphocyte proliferation was assessed to observe the influence of centella asiatica extracts and to assess cell cycle and the concentration of intracellular [Ca2+]i in spleen lymphocytes.Methods1.Centella asiatica was isolated with alcohol, different organic solvent and Sephadex-LH20 gel column chromatography was applied for further purification.2. The methods of scavenging capacity against radical of OH·, DPPH·were used to evaluate the antioxidant activity of centella asiatica extracts.3. The spleen lymphocyte proliferation model of mice was established to observe the influence of centella asiatica extracts.4. Flow cytometry was performed to assess cell cycle, Application Fluo-3/AM fluorescence probe combined Flow cytometry analysis of the concentration of intracellular [Ca2+]i in spleen lymphocytes of extracts from centella asiatica.Results1. The A,B,C,D four fractions were gained by furthering extracted form the crude extracts of centella asiatica.The further purification extract D from centella asiatica was obtained and confirm extract D inhibit DPPH·, OH·ability most evident. The active ingredient that it is extract D was identified as flavone.2. Only D extract from centella asiatica could suppress lymphocytes proliferation.3. D extract from centella asiatica could decrease the concentration of the intracellular [Ca2+]i of spleen lymphocytes in a dose-dependentmanner.4. D extract from centella asiatica could arrest cell cycle at S phage and G2/M.ConclusionsThe activite ingredient extract from centella asiatica have possesed antioxidant activity and it was identified as flavonoid. The ability of centella asiatica for lowing intracellular [Ca2+]i and arresting cell cycle at S phage and G2/M of spleen lymphocytes could be responsible for its anti-proliferative activites.