Trifolium pratense is known to have antioxidant, anti-inflammatory, antilipidemic, hepatoprotective and anticarcinogenic effects. The purpose of this investigation is to investigate the possible effects of Trifolium pratense on purine degradation rate.
Adenosine degradation in the biological tissue was monitored by analyzing uric acid levels. While 27.27 % increase at uric acid levels was determined in control samples, 130,05 % increases were found for extract-treated samples.
The research has shown that Trifolium pratense may exhibit anticarcinogenic activity by causing purine degradation.
Cancer takes an attention as a worldwide health problem . In addition to radiotherapy and chemotherapy, natural remedies draw attention to cancer treatment .
Trifolium pratense is known to have antioxidant, anti-inflammatory, antilipidemic, hepatoprotective and anticarcinogenic effects due to its active substances like Taxifolin and quercetin . It is also used for reducing menopausal symptoms like hot flushes .
Purines and pyrimidines are precursors of DNA synthesis. Adenosine is a purine nucleoside which also plays a role in energy transfer and signal transduction. Nucleotides are synthesized in both de novo and salvage pathways. Products released after nucleic acid and nucleotide cofactor catabolism constitutes purine bases which are used in nucleotide synthesis at salvage pathways . Degradation of these precursors by other factors can deplete nucleotide pool and decrease DNA synthesis.
We aimed to investigate possible effects of Trifolium pratense on purine degradation rate. For this purpose, adenosine degradation in the biological tissue was monitored by analyzing uric acid levels during incubation.
Fresh cattle liver was obtained from a local slaughterhouse. Liver tissues were stored at -80oC till analysis. Before the assays, liver tissue was homogenized by DIAX 900 (Heidolph, Germany) in saline solution (%10 w/v). Then, the homogenates were centrifuged at 3000 rpm for 20 minutes by Harrier 18/80 centrifuge (MSE, London, UK). The clear supernatant fraction of liver homogenate was then separated and used in the assays.
Aqueous extract of Trifolium pratense was prepared at the concentration of %10(w/v) with distilled water.
Adenosine solution was prepared at a concentration of 0,5mM.
Adenosine solution (0,1 mL) was added into 1 mL supernatant fraction of liver homogenate. Then, 0,5 mL aqueous extract of Trifolium pratense was added into the mixture. For control samples, 0,5 mL distilled water was used instead of an aqueous extract of Trifolium pratense. Mixtures of the control and treated samples were analyzed for uric acid determination immediately
and after 20 hours of incubation at 37oC. Modified alkaline
phosphotungstate method was utilized for determination
of uric acid concentration . Absorbances were measured
spectrophotometrically by using Helios alpha Ultraviolet/Visible
Spectrophotometer (Unicam, Cambridge, UK).
Preincubation and postincubation uric acid levels were
compared by using paired sample T-test. After incubation,
statistically significant differences were found for both control
and treated samples (Table 1). While 27.27 % increase at uric acid
levels was determined in control samples, 130,05 % increases
were found for extract-treated samples. The differences were
found statistically significant (Table 1).
Trifolium pratense contains a high amount of polyphenolic
substances which exhibit antioxidant effects. Most widely
studied feature of Trifolium pratense is its estrogenic effects and
usage in postmenopausal women [7,8].
There is insufficient knowledge about anticarcinogenic
activities of Trifolium pratense in literature. Jarred et al 
showed that Trifolium pratense-derived dietary isoflavones led
to apoptosis in prostate carcinoma tissue .
In our study, we investigated adenosine degradation by
analyzing uric acid levels before and after incubation. Uric acid
levels were increased significantly in both treated and control
samples. While %27 increase at uric acid levels was detected
for control samples, %130 increase was determined for samples
treated with Trifolium pratense as summarized in (Table 1).
These results suggest that Trifolium pratense activate adenosine
It is known that purine and pyrimidine bases are utilized
in nucleotide synthesis at salvage pathways . Degradation of
these precursors can inhibit DNA synthesis in owing to substrate
deficiency in the enzymatic process. Cell generation may halt
due to DNA synthesis reduction. Rapid propagation of cancerous
tissues can be decreased by inhibiting DNA synthesis. Trifolium
pratense may exhibit anticarcinogenic activity by causing purine
degradation. Further studies are required to establish possible
anti-carcinogenic effects of Trifolium pratense.