The Effect of Medium in Properties and Activity of Acid Phosphatase (ACP) in the Infective Form of Leishmania Major Promastigotes

• Main Authors: Amir Navabi, Simindokht Soleimanifard
• Format: Article
• Language: fas
• Published: Vesnu Publications 2014-10-01
• Series: مجله دانشکده پزشکی اصفهان
• Subjects: Leishmania major; Media culture; Acid phosphatase; Virulence
• Online Access: http://jims.mui.ac.ir/index.php/jims/article/view/3367

Background: The wet cutaneus leishmaniasis caused by Leishmania major infection is one of the common skin diseases in many parts of the world. Reports indicate that some properties of Leishmania, such as time of infection, is affected of medium. Respecting the use of artificial medium in the majority of the studies on Leismania, and the role of acid phosphatase (ACP) in the pathogenesis of promastigotes, in this research, we studied the effects of medium in properties of acid phosphatase and time of infecting in Leishmania major promastigotes. Methods: In this cross-sectional study, to culture promastigotes, Leishmania major (MRHO/IR/75/ER) from previously infected Balb/c mice was transferred to NNN (Novy-MacNeal-Nicolle) and RPMI1640 (Roswell Park memorial institute) medium. Growth curve was generated and stationary stages were divided. Frozen promastigotes of each medium were homogenized using sodium acetate and Triton-X-100 and acid phosphatase was measured via calorimetric assay. Findings: Stationary parasites were collected in NNN and RPMI-1640 medium in seventh and fifth day, respectively. The rate of acid phosphatase activity was determined as 1.02±0.08 in NNN and 1.8 ± 0.01 in RPMI-1640. Also, Km (Michaelis–Menten) and Vmax (Maximum velocity) of this enzyme was 105.26 ± 1.11 µM and 98.37 ± 1.48 µM/min/mg protein in NNN and 106.39 ± 1.14 µM and 98.04 ± 0.96 µM/min/mg protein in RPMI1640. Conclusion: Despite the infecting time in promastigotes, rate of agglutination, rate of kinase synthesis and virulent are different in two mediums. It seems that there are no significant differences in properties of acid phosphatase in stationary promastigotes in the two mediums.