Abstract
In this paper to isolate and study the properties of the cyclooxygenase-2 (EC: 1.14.99.1) enzyme in the blood of a patient suffering from rheumatoid arthritis and study the effect of natural products of the Soapwort on the activity of purified enzyme. The study involves taking 30 ml of blood from an adult woman 40 years old, who suffers from rheumatoid arthritis disease for 13 years. Serum is separated and subjected to a series of purification processes including: precipitation by ammonium sulfate, filtration by centrifugation radiator, dialysis in presence of ammonium bicarbonate, separation using the technology of ion exchange, lipholization and then estimating approximate molecular weight of the enzyme using gel filtration technique and sodium dodecyl sulfate (SDS)-page polyacrylamide gel electrophoresis. The study also includes isolating the natural products of Soapwort plant and study the effect of isolated natural products on the activity of the purified enzyme. The result of the study indicates that cycloxygenase-2 has an approximate molecular weight of 71.5 kDa and that the extracted oil of the Soapwort has a negative impact on the activity of the enzyme (r= -0.824; P=0.006), while flavonoids and Saponin have no such impact (r= -0.565; P=0.113; r= -0.634; P=0.067 respectively).
Highlights
Cyclooxygenase (COX, called prostaglandin H synthase or PGHS), (EC: 1.14.99.1), is a key enzyme in the synthesis of prostanoids [1, 2]
The important feature of synovial cells is that the cells stimulate COX-2 expression [6], for this reason, the blood sample that applies in the current study is selected from a patient with rheumatoid arthritis because in the case of rheumatoid arthritis the inflammation gets out of control and the immunity system becomes more active and increases the concentration of the factors that induce the secretion of the COX-2 enzyme such as lipopolysaccharides [7] cytokines [8], tumor necrosis factor (TNF) [9]
There occurs a failure of acute inflammation to resolve may predispose to: autoimmunity, chronic dysplastic inflammation, excessive tissue damage and occurs dramatic increase of COX2 expression [14], and because COX-2 is one of the most important enzymes that is responsible for inflammation, the current study aims to purify COX-2 from a blood of a patient with rheumatoid arthritis and study the effect of Soapwort natural products on the activity of purified enzyme
Summary
Cyclooxygenase (COX, called prostaglandin H synthase or PGHS), (EC: 1.14.99.1), is a key enzyme in the synthesis of prostanoids [1, 2]. Inflammatory mediators, including tumor necrosis factor (TNF) and interleukin (IL-l), release from the inflammatory cells in response to various stimuli activate fibroblast-like synovial cells. The important feature of synovial cells is that the cells stimulate COX-2 expression [6], for this reason, the blood sample that applies in the current study is selected from a patient with rheumatoid arthritis because in the case of rheumatoid arthritis the inflammation gets out of control and the immunity system becomes more active and increases the concentration of the factors that induce the secretion of the COX-2 enzyme such as lipopolysaccharides [7] cytokines [8], tumor necrosis factor (TNF) [9]
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