Mohamad Sadikin
Department of Biochemistry and Molecular Biology, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia

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The increased of carbonic anhydrase in liver tissue of rat induced by chronic systemic hypoxia Rahmawati Ridwan; Febriana Catur Iswanti; Mohamad Sadikin
Acta Biochimica Indonesiana Vol. 1 No. 1 (2018): Acta Biochimica Indonesiana
Publisher : Indonesian Society for Biochemistry and Molecular Biology

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.32889/actabioina.v1i1.1

Abstract

Background: Carbonic anhydrases (CAs) are metalloenzymes which catalyze the reversible hydration/dehydration reaction of CO2, in order to maintain the cell homeostasis. These enzymes are found in various tissues and involve in a number of different physiological processes, including ion transport, acid-base balance, bone formation, and gluconeogenesis. Objective: To examine the specific activity of CA and to observe the liver tissue respond to oxidative stress by measured the malondialdehyde (MDA) concentration, in rat liver tissue induced by chronic systemic hypoxia for 1, 3, 5, 7, and 14 days of hypoxia. Results: The study showed that the activity of CA induced by chronic systemic hypoxia significantly increasing at early exposure to the hypoxic condition, at day 1 and days 3 of hypoxia (0.281 and 0.262 nmol/mg protein/minute compared to control 0.155 nmol/mg protein/minute) (p<0.05). No statistical difference at treatments of hypoxia 5, 7, and 14 days. The concentration of MDA also increased significantly on day 3 of liver tissue hypoxia (0.013 nmol/mg compared to control 0.009 nmol/mg liver tissue) (p<0.05), and no statistical differences at day 1, 5, 7, and 14 days of hypoxia. Conclusion: There was damage of membrane cells affected by oxidative stress in the liver tissue of rats induced by chronic systemic hypoxia.
Carbon tetrachloride administration induces the expression of hypoxia inducible factor-1alpha in rat liver Lindi G Haritsyah; Mohamad Sadikin; Sri Widia Jusman
Acta Biochimica Indonesiana Vol. 1 No. 1 (2018): Acta Biochimica Indonesiana
Publisher : Indonesian Society for Biochemistry and Molecular Biology

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.32889/actabioina.v1i1.3

Abstract

Background: There is now increasing evidence that HIF-1 is also responsive to a variety of non-hypoxic stimuli. However, the mechanisms by which these non-hypoxic stimuli induce HIF-1α are not completely known, yet, although some evidence points to a role of ROS as messengers regulating HIF activity. Objective: To determine the expression of HIF-1α in liver rat tissue induced by carbon tetrachloride under normoxic conditions, with or without N-acetylcysteine protection. Methods: Twenty-five male Sprague-Dawley rats were divided into 5 groups: normal control rats, normal rats orally administered with coconut oil (1 mL/200 g body weight) for 1 day, rats orally administered with CCl4 (0.55 mg/g body weight) for 1 day, rats injected i.v. with NAC (0.15 mg/g body weight) for 8 days and then orally administered with CCl4 (0.55 mg/g body weight) for 1 day, rats orally administered with CCl4 (0.55 mg/g body weight) for 1 day and then injected i.v. with NAC (0.15 mg/g body weight) for 2 days. The expression of HIF-1α mRNA was measured by real-time RT-PCR using the Livak method. The expression of HIF-1α protein was measured by ELISA assay. Results: The highest HIF-1α mRNA and protein expression found in the group treated by CCl4 and then was gradually lowered in the pre-NAC group, post-NAC group, control group, and last, in the oil group. Conclusion: Our study shows the effect of CCl4-treated rats under normoxic conditions increased the mRNA and protein HIF-1α. NAC post-treatment provide a better protective effect compared with NAC pre-treatment
Palladium (II) chloride (PdCl2) spectrophotometry to determine lipoic acid concentration in plasma and leukocytes Novian Agni Yudhaswara; Ani Retno Prijanti; Mohamad Sadikin
Acta Biochimica Indonesiana Vol. 3 No. 1 (2020): Acta Biochimica Indonesiana
Publisher : Indonesian Society for Biochemistry and Molecular Biology

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.32889/actabioina.v3i1.29

Abstract

Background: Lipoic acid is a substance contained in intra and extracellular that act as a coenzyme of Pyruvate Dehydrogenase, also as an antidote, chelating agent and antioxidant. Measurement of lipoic acid is needed to determine the amount of lipoic acid that performs its functions either as a coenzyme or an antioxidant. Besides, this measurement requires a special tool such as High Performance Liquid Chromatography (HPLC) and a process that is available in rural or simple laboratories. Objective: A common and easy tool such as a spectrophotometer was conducted and could expected to be a tool of lipoic acid determination in body fluid such as plasma. Methods: Measurement of lipoic acid using spectrophotometry with UV methanol and visible PdCl2 has been tested and compared to HPLC measurement that was valid and reliable in drug measurement or pharmaceutical preparations. Results: Determination of lipoic acid in plasma and leukocytes using PdCl2 produced replicable, reliability and valid result, with high accuracy, precision and was not different from lipoic acid measurement using HPLC, p=0.99. While UV methanol was different compare to HPLC p =0.0001 or was not valid. Conclusion: The measurement of lipoic acid using PdCl2 visible method can be applied to determine the levels of lipoic acid (LA) and DHLA in plasma and equal to HPLC result.