IN VITRO ANTI-INFLAMMATORY ACTIVITY TEST OF TINOCRISPOSIDE AND FREEZE-DRIED AQUEOUS EXTRACT OF TINOSPORA CRISPA STEMS ON HUMAN RED BLOOD CELL BY INCREASING MEMBRANE STABILITY EXPERIMENT
Journal Title: Asian Journal of Pharamceutical and Clinical Research - Year 2019, Vol 12, Issue 5
Abstract
Objective: This study was aimed to evaluate the anti-inflammatory effect of isolated tinocrisposide and freeze-dried aqueous extract of Tinospora crispa stems on human red blood cell (HRBC) by increasing membrane stability in vitro models. Methods: Anti-inflammatory effect of tinocrisposide and FDAETCS was evaluated by in vitro HRBC membrane stabilization method. The study was separated into two steps which were a hemolytic and a membrane stabilization experiment. The hemoglobin that was released throughout the damaged erythrocytes membrane was then quantified at the wavelength of (λ) 560 nm. Results: The hemoglobin in the HBRC supernatant that treated with tinocrisposide at concentration of 100, 200, 400, 600, 800, and 1000 μg/ml showed an absorbance at λ 560 nm of 0.060, 0.061, 0.071, 0.072, 0.075, and 0.0793, respectively, and the calculated hemolysis percentage was 0.032, 0.097, 1.203, 1.236, 1.641, and 2.079%, respectively. We found a linear correlation between concentration and hemolytic activity of tinocrisposide, with regression equation, y=0.0023x−0.1312 (r=0.929). Meanwhile, the HBRC supernatant that treated with FDAETCS at concentration of 100, 200, 400, 600, and 800 μg/ml showed an absorbance at λ 560 nm of 0.063, 0.064, 0.066, 0.067, and 0.077, respectively, and revealed the hemolytic percentage of 0.347, 0.473, 0.693, 0.992, and 1.896%, respectively. It also gave a linear correlation between FDAETCS concentration and hemolytic activity percentage, with regression equation, y=0.002x+0.0222 (r=0.895). Moreover, in HRBC membrane stability experiment, tinocrisposide concentration of 100, 200, 400, 800, and 1000 μg/ml gave absorbance at λ 560 nm of 0.818, 0.808, 0.798, 0.789, 0.773, and 0.761, respectively, and calculated HRBC membrane stabilization activity as much as 5.437, 6.533, 7.707, 8.748, 10.597, and 12.100%, respectively. Meanwhile, the positive control ibuprofen 25 μg/ml only exerted the membrane stability of 5.620%. It was found a linear correlation between tinocrisposide concentration and membrane stability percentage, with the regression equation, y=0.0072x+4.8312 (r=0.9932). Treated FDAETCS in the concentration of 100, 200, 400, and 800 μg/ml gave the absorbance at λ 560 nm of 0.802, 794, 0.777, 0.791, and 0.792, with stability membrane percentage of 7.283, 8.208, 10.944, 8.555, and 8.401%, respectively. It can be seen that the FDAETCS concentrations and its hemolytic percentage showed a parabolic relationship, which gave a maximum at a concentration of the extract of 400 mg/ml with membrane stabilizing of 10.944%. Conclusion: It can be concluded that tinocrisposide and FDAETCS have an anti-inflammatory activity by increase the membrane stability of lysosome cell that has equal physiological properties with erythrocytes membrane cell and it has no hemolytic activity.
Authors and Affiliations
ADEK ZAMRUD ADNAN , FITHRIANI ARMIN , IKHWAN RESMALA SUDJI , MEDIOLA DWI NOVIDA , DEWI IMELDA ROESMA , HANA AFIFAH ALI , ANNISA FAUZANA
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