Kandikattu Hemanth Kumar, Rachitha P, Krupashree K, Jayashree G V, Abhishek Virat, Khanum Farhath
Biochemistry and Nanosciences Discipline, Defence Food Research Laboratory, Mysore 570011, Karnataka, India.
Food Engineering and Packaging Division, Defence Food Research Laboratory, Mysore 570011, Karnataka, India.
Pathophysiology. 2015 Dec;22(4):165-73. doi: 10.1016/j.pathophys.2015.07.001. Epub 2015 Jul 28.
In the present investigation, we identified the phytochemical constituents of total oligomeric flavonoid fraction (TOF) of Cyperus rotundus by LC-ESI-MS/MS and also demonstrated its antihemolytic effects against 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) induced hemolysis of rat erythrocytes. Our results of TOF extract exhibited DPPH, metal chelating, ABTS, NO and hydroxyl radical scavenging activities with an IC50 values of 23.72±1.6, 52.45±2.88, 9.8±0.42, 6.5±0.33 and 120±6.83μg/ml respectively, whereas total antioxidant and reducing power activities were 194±12.5μg GAE/mg extract and 145±8.3μg AAE/mg extract. The extract showed potent inhibitory activity against AAPH induced plasmid DNA damage, protein oxidation and lipid peroxidation. The TOF extract mitigates AAPH induced hemolysis and exhibits ∼50% antihemolytic activity. TOF pretreatment also preserved morphology of erythrocytes as observed and measured by light microscope and atomic force microscope analysis. Furthermore, the TOF fraction effectively inhibited AAPH induced LDH release, ROS generation and lipid peroxidation. Taken together, our data demonstrate the antihemolytic activity of C. rotundus against AAPH induced oxidative stress of erythrocytes, and was associated with the decrease in oxidative stress, cellular damage and protection of macromolecules. In conclusion, the effects might be correlated with high content of flavonoids and polyphenols identified in C. rotundus. This suggests the clinical application of TOF fraction of C. rotundus against ROS induced cell death.
在本研究中，我们通过液相色谱-电喷雾串联质谱（LC-ESI-MS/MS）鉴定了香附总寡聚黄酮部分（TOF）的植物化学成分，并证明了其对2,2'-偶氮二异丁脒二盐酸盐（AAPH）诱导的大鼠红细胞溶血的抗溶血作用。我们的TOF提取物结果显示出二苯基苦味酰基自由基（DPPH）清除、金属螯合、2,2'-联氮-双-（3-乙基苯并噻唑啉-6-磺酸）二铵盐（ABTS）清除、一氧化氮（NO）清除和羟基自由基清除活性，其半数抑制浓度（IC50）值分别为23.72±1.6、52.45±2.88、9.8±0.42、6.5±0.33和120±6.83μg/ml，而总抗氧化活性和还原能力分别为194±12.5μg没食子酸当量（GAE）/mg提取物和145±8.3μg抗坏血酸当量（AAE）/mg提取物。该提取物对AAPH诱导的质粒DNA损伤、蛋白质氧化和脂质过氧化显示出强大的抑制活性。TOF提取物减轻了AAPH诱导的溶血，并表现出约50%的抗溶血活性。通过光学显微镜和原子力显微镜分析观察和测量发现，TOF预处理还保留了红细胞的形态。此外，TOF部分有效抑制了AAPH诱导的乳酸脱氢酶（LDH）释放、活性氧（ROS）生成和脂质过氧化。综上所述，我们的数据证明了香附对AAPH诱导的红细胞氧化应激具有抗溶血活性，并且与氧化应激的降低、细胞损伤的减轻和大分子的保护有关。总之，这些作用可能与香附中鉴定出的高含量黄酮类化合物和多酚类化合物有关。这表明香附TOF部分在临床上可用于对抗ROS诱导的细胞死亡。
