Volume 22, Issue 4, Pages (December 2015)

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Volume 22, Issue 4, Pages 165-173 (December 2015) LC–ESI-MS/MS analysis of total oligomeric flavonoid fraction of Cyperus rotundus and its antioxidant, macromolecule damage protective and antihemolytic effects Hemanth Kumar Kandikattu, P. Rachitha, K. Krupashree, G.V. Jayashree, Virat Abhishek, Farhath Khanum Pathophysiology Volume 22, Issue 4, Pages 165-173 (December 2015) DOI: 10.1016/j.pathophys.2015.07.001 Copyright © 2015 Elsevier B.V. Terms and Conditions

Fig. 1 LC–ESI-MS/MS of hydroalcoholic fraction of Cyperus rotundus. Pathophysiology 2015 22, 165-173DOI: (10.1016/j.pathophys.2015.07.001) Copyright © 2015 Elsevier B.V. Terms and Conditions

Fig. 2 Chemical structures of the compounds of Cyperus rotundus identified by LC–ESI-MS/MS analysis. Pathophysiology 2015 22, 165-173DOI: (10.1016/j.pathophys.2015.07.001) Copyright © 2015 Elsevier B.V. Terms and Conditions

Fig. 3 (a) The protective effect of C. rotundus on 2,2′-azobis(2-amidinopropane) dihydrochloride induced plasmid DNA damage analysis by agarose gel electrophoresis. (b) The protective effect of C. rotundus on 2,2′-azobis(2-amidinopropane) dihydrochloride induced protein oxidation of bovine serum albumin analysed by polyacrylamide gel electrophoresis. (ii), The densitometric analysis of protein oxidation by NIH Image J software. *P<0.05 versus AAPH treated BSA. Pathophysiology 2015 22, 165-173DOI: (10.1016/j.pathophys.2015.07.001) Copyright © 2015 Elsevier B.V. Terms and Conditions

Fig. 4 (a) The hemolytic effects of different concentrations of 2,2′-azobis(2-amidinopropane) dihydrochloride on erythrocytes. The results were analyzed by one-way ANOVA. *P<0.05 versus AAPH treated cells (n=5). (b) The antihemolytic effect of TOF fraction on 2,2′-azobis(2-amidinopropane) dihydrochloride induced hemolysis. The results were analyzed by one-way ANOVA. *P<0.05 versus AAPH treated cells (n=5). Pathophysiology 2015 22, 165-173DOI: (10.1016/j.pathophys.2015.07.001) Copyright © 2015 Elsevier B.V. Terms and Conditions

Fig. 5 (A) The morphological observation of erythrocytes by light microscope (Olympus, Japan). (a) Control cells (b) erythrocytes treated with 100mM 2,2′-azobis(2-amidinopropane) dihydrochloride for 4h (c) erythrocytes treated with 50μg/mL TOF followed by 100mM 2,2′zobis(2-amidinopropane) dihydrochloride treatment for 4h. (B) The morphological observation and measurement of size and height of erythrocytes by Atomic force microscope analysis (NT-MDT, Russia). (a) Control cells (b) Erythrocytes treated with 100mM 2,2′-azobis(2-amidinopropane) dihydrochloride for 4h (c) Erythrocytes treated with 50μg/mL TOF followed by 100mM 2,2′-azobis(2-amidinopropane) dihydrochloride treatment for 4h. Pathophysiology 2015 22, 165-173DOI: (10.1016/j.pathophys.2015.07.001) Copyright © 2015 Elsevier B.V. Terms and Conditions

Fig. 6 Cytotoxic effect of 2,2′-azobis(2-amidinopropane) dihydrochloride and its protective effect by TOF on erythrocytes analyzed by LDH assay. The results were analyzed by one-way ANOVA. *P<0.05 versus AAPH treated cells (n=5). Pathophysiology 2015 22, 165-173DOI: (10.1016/j.pathophys.2015.07.001) Copyright © 2015 Elsevier B.V. Terms and Conditions

Fig. 7 (a) Estimation of ROS generation in erythrocytes on treatment with 2,2′-azobis(2-amidinopropane) dihydrochloride/TOF using a fluorescent probe 2′,7′-DCFH2DA. The results were analyzed by one-way ANOVA. #P<0.05 versus control cells. *P<0.05 versus AAPH treated cells (n=5). (b) Estimation of lipid peroxidation in erythrocytes on treatment with 2,2′-azobis(2-amidinopropane) dihydrochloride/TOF by MDA method. The results were analyzed by one-way ANOVA. #P<0.05 versus control cells. *P<0.05 versus AAPH treated cells (n=5). Pathophysiology 2015 22, 165-173DOI: (10.1016/j.pathophys.2015.07.001) Copyright © 2015 Elsevier B.V. Terms and Conditions