Purification of paraoxonase enzyme from the sera of patients with Behcet’s disease and analyzing the effects of the drugs containing imuran (azathioprine), prednisolone (methylprednisolone) and colchium (colchicine)

Investigation of Paraoxonase (PON) Enzyme Activities in Behçet’s Disease

This comprehensive study analyzes serum samples from 50 patients diagnosed with Behçet’s disease and 20 healthy volunteers to evaluate enzyme behavior. The research is structured into three distinct phases, focusing on the activity, purification, and inhibition profiles of the Paraoxonase (PON) enzyme. By comparing patient data with a control group, the study provides critical insights into the biochemical markers of the disease.

Determination of PON Activities and Enzyme Purification

In the initial phase of the research, Paraoxonase (PON) activities were measured in the serum samples of both the 50 Behçet’s patients and the 20 healthy individuals. This baseline measurement serves as a foundation for understanding the enzymatic differences between the two groups.

Advanced Purification and Characterization Techniques

Following the activity measurements, serum samples from the patient group were pooled for a detailed purification process. The PON enzymes were purified using a Sepharose-4B-L-tyrosine tyrosine-1-naphtylamine affinity column. To characterize the purified enzymes, several kinetic parameters were established, including:

• Optimum temperature and optimum pH levels.
• Vmax and Km values of the pure enzymes.
• Electrophoretic mobility assessment via SDS-PAGE.

This exact purification and characterization procedure was also applied to the serum samples of the healthy control group to ensure a standardized comparison.

In Vitro Effects of Therapeutic Drugs on PON Enzymes

The final stage of the study investigated the in vitro effects of specific drugs commonly used in the treatment of Behçet’s disease. The research focused on the interactions between the PON enzymes and the following pharmacological agents:

• Azathioprine
• Methylprednisolone
• Colchicine

Analysis of Inhibition Kinetics

For the drugs demonstrating in vitro inhibition effects, precise biochemical constants were calculated for both the patient and control groups. The study determined the IC50 values and Ki constant values to quantify the potency of the inhibition. Furthermore, the specific inhibition types were identified for azathioprine, methylprednisolone, and colchicine, providing a detailed map of how these treatments interact with the Paraoxonase enzyme system.