Abstract

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Background: To determine the effect caused by the methanolic extract of Trichosanthes cucumerina seed on experimentally increased PSA in adult male Wistar rats. Place and Duration of Study: Department of Anatomy, Faculty of Basic Medical Science, Olabisi Onabanjo University, Ikenne, Ogun State Nigeria, for 6weeks.. Methodology: We included 20 adult male Wistar rats, of 4 groups (Normal control NCTRL, Hormone treated control HTC, High dose of extract HDT/C, Low dose of extract LDT/C) with each group comprising of 5 animals each, with individual rats weighing between 150g-320g. The increased PSA value was achieved by the simultaneous induction of 400ng/ml of Estradiol and 1250ng/ml of Testosterone (with respect to the subject’s body weight), which were administered via the inguinal region for a period of 3 weeks (in alternate days). After the period of hormonal administration, two of the subjects from groups NC and HTC were sacrificed; blood samples were taken from the heart apex in other to test for the PSA values. Furthermore, with the methanolic extract of the plant’s seed, the animals grouped in HD/TC and LDT/C was treated with respect to their body weight, for a period of 3 weeks. After which, the animals were sacrificed, blood samples were again taken to obtain the PSA value. All results were expressed as Mean ± Standard Deviation (S.D) for each group. All grouped data were statistically evaluated using SPSS 15.0 software. Hypothesis testing methods included the independent – samples t–test. Statistical significance was set at p<0.05. Result: The experimentally induced increased in PSA was able to be restored back to normal due to the effect of the methanolic extracts of Trichosanthes cucumerina seed Conclusion: The combined administration of Testosterone and Estradiol caused an increase in the PSA values which were reversed after the dosage (high and low) administration of the extract.

Introduction

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Prostate-specific antigen (PSA) also known as gamma-seminoprotein or kallikrein-3 (KLK3) is a glycoprotein that in humans is encoded by the KLK3 gene. [1] KLK3 is a member of the kallikrein-related peptidase family secreted by the epithelial cells of the prostate gland. PSA is produced for the ejaculate where it liquefies the semen in the seminal coagulum and allows sperm to swim freely.[2] It is also believed to be instrumental in dissolving the cervical mucus, allowing the entry of sperm.[3]PSA is present in small quantities in the serum of men with healthy prostates, but is often elevated in the presence of prostate cancer and in other prostate disorders. In those with prostate cancer, rising levels of PSA over time are associated with both localized and metastatic prostate cancer (CaP). Prostate test screening is controversial and may lead to unnecessary, even harmful, consequences in some patients. Prostate-specific antigen (PSA, also known as kallikrein III, seminin, semenogelase, ?-seminoprotein and P-30 antigen) is a 34 kD glycoprotein produced almost exclusively by the prostate gland. It is a serine protease (EC 3.4.21.77) enzyme, the gene of which is located on the nineteenth chromosome in humans.[4] PSA is normally present in the blood at very low levels. The reference range of less than 4 ng/mL for the first commercial PSA test, the Hybritech Tandem-R PSA test released in February 1986, was based on a study that found 99% of 472 apparently healthy men had a total PSA level below 4 ng/mL—the upper limit of normal is much less than 4 ng/mL.[5] Increased levels of PSA may suggest the presence of prostate cancer. However, prostate cancer can also be present in the complete absence of an elevated PSA level, in which case the test result would be a false negative.[6] Obesity has been reported to reduce serum PSA levels.[7] Delayed early detection may partially explain worse outcomes in obese men with early prostate cancer.[8] PSA levels can be also increased by prostatitis, irritation, benign prostatic hyperplasia (BPH), and recent ejaculation,[9][10] producing a false positive result. Digital rectal examination (DRE) has been shown in several studies [11] to produce an increase in PSA. However, the effect is clinically insignificant, since DRE causes the most substantial increases in patients with PSA levels already elevated over 4.0 ng/mL. The "normal" reference ranges for prostate-specific antigen increase with age, as do the usual ranges in cancer:[12] In women, PSA is found in female ejaculate at concentrations roughly equal to that found in male semen.[13] Other than semen and female ejaculate, the greatest concentrations of PSA in biological fluids are detected in breast milk and amniotic fluid. Low concentrations of PSA have been identified in the urethral glands, endometrium, normal breast tissue and salivary gland tissue. PSA also is found in the serum of women with breast, lung, or uterine cancer and in some patients with renal cancer.[14] Snake gourd (Trichosanthes cucumerina) belongs to the family Cucurbitaceae mostly consumed as vegetable, but it may grow throughout the year except extreme winter. It is a popular vegetable with moderately high nutritive value. The total production of snake gourd during 2003-2004 was 136000 tons on the area of 1, 59,000 acres of land. [15] This figure indicates the low yield potentiality of our cultivars. It is commonly called as snake gourd, viper gourd, snake tomato or long tomato. The fruit is usually consumed as a vegetable due to its good nutritional value. The plant is richly constituted with a series of chemical constituents like flavonoids, carotenoids, phenolic acids which makes the plant pharmacologically and therapeutically active. Its Fruit is regarded as anthelmintic, vomitive [16] antidia-betic [17] for boil [18]. Seeds are anthelmintic,and anti fibrile [19]. Root is used as purgative and tonic. The pharmacological activities of cucurbitacin containing plants have been known since ancient times. Cucurbitacins are particularly known in folk medicine for their strong purgative, anti-inflammatory, and hepatoprotective activi-ties [20].

Methods

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Management Adult male rats, weighing between 150g-320g were obtained from the animal house of the Department of Zoology, University of Ibadan, Nigeria. They were housed in the animal house provided by the Department of Anatomy, Olabisi Onabanjo University, where they had access to food, water, and air. Their environment was well cleaned to avoid infection of any kind upon the animals. After a week of acclimatization, the animals were induced with specific dosage of Estradiol and Testosterone (both diluted in corn oil respectively). The hormones were given based on the animal’s body weight, and the route of administration being the inguinal region. After 3weeks of induction, some of the subjects were sacrificed, blood samples taken, to test for the prostate specific antigen. Others were then treated with the methanolic extracts of Trichosanthes cucumerina (in proportion to their body weight), diluted in corn oil. The treatment lasted for 3weeks after which the animals were sacrificed. Drug Administration The hormones were diluted in corn oil into 400ng/ml and 1250ng/ml of estradiol and testosterone respectively. Hormones were administered exogenously via the inguinal region for three weeks, at thrice a week in alternate days. [21] The extracts were given at 0.2mg/ml and 0.1mg/ml of high and low dosages [21]The testosterone was manufactured by Green Field Pharm. (JIANG SU) Co., Ltd, China while Estradiol- manufactured by Medipharm (Pvt.) Ltd., 108-Kotlakhpat Industrial Estate, Lahore. Method of Extraction Six ripe fruits of Trichosanthes cucumerina were gotten from Ayepe, a local town in ijebu-remo side of Ogun State, Nigeria. The seeds were obtained from the fruit, washed in clean water; sun dried for three days, the coat peeled off, 40g of the seeds were weighed, grinded into fine powder and finally socked in 100ml of methanol. The solution was filtered after 48 hours while the filtrate was concentrated using the rotary evaporator; volume of filtrate obtained =30.6ml, weight of residue left =32g; weight of the seeds dissolved in the filtrate= 8g; volume of filtrate (oil obtained from the seeds) after evaporating = 2.5ml. Therefore; extracts concentration = mass/volume = 8/2.5 = 3.2g/ml Blood Test Animals were anaesthetized with chloroform in closed chamber. The thoracic vertebrate was opened under aseptic condition, blood were taken carefully through the apex of the heart and taken to the laboratory for PSA analysis.

Results

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1. For the Normal Control group (NCTRL), 5 rats were used and the Mean ± S.D was = 3.75 ± 1.71.
2. For the Hormone treated Control group (HCTRL), 5 rats were used and the Mean ± S.D was = 8.75 ± 0.96.
3. For the High Dose of Plant Extract group (HD T/C), 5 rats were used and the Mean ± S.D was = 2.75 ± 0.96 c, b.
4. For the High Dose of Plant Extract group (HD T/C), 5 rats were used and the Mean ± S.D was = 6.25 ± 2.63 c, a. a- Means there is No significant difference between H CTRL when compared with LD T/C. c- Means there is No significant difference between N CTRL when compared with HD T/C & LD T/C. b- Means there is significant difference between H CTRL when compared with HD T/C.

Discussion

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It could be observed from this study that in the normal control group of rat (NCTRL), the PSA value was observed to be normal since the animals were not treated at all. Whereas in the hormone treated group (HCTRL), there was a significant increase in the PSA. [22]. However, in groups treated with low dose of Trichosanthes cucumerina (LDT/C) there was a gradual reversal to the normal (6.25 ± 2.63) while in the group administered with High dose of Trichosanthes Cccumerina (HDT/C) showed a complete reversal to the normal level at 2.75 ± 0.96 . This study provides a model for demonstrating the mechanism of prostatic enlargement in the aging prostate, which results in increase in PSA value. It suggests that there is some level of conversion of Testosterone to estradiol-17-beta in the aging prostate by the enzyme aromatase and the estrogen so produced though at low level synergizes the effect of T and its metabolite dihydrotestosterone DHT in the prostate thereby leading to prostatic hypertrophy

Conclusion(s)

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The combined administration of Testosterone and Estradiol caused an increase in the PSA values which were reversed after the dosage (high and low) administration of the extract.

Abbreviation(s)

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T/C- Trichosanthes cucumerina

METC- Methanolic extract of Trichosanthes cucumurina seed MEAN ± SD- Mean plus OR minus Standard Deviation p<0.05- Statistical significance was set at p< 0.05. t-test- samples t–test

Acknowledgement(s)

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I wish to express my sincere gratitude to the co-authors as well as the technical staff of the Anatomy Department OOU, Ikenne Ogun-State Nigeria, for their support in execution of this work.

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Recommendation

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The solution to this problem is to research on substances which can selectively block estrogen receptors without affecting the androgen receptors in the aging prostate, as this may offer solution to problem of prostatic enlargement and hypertrophy.

Source(s) of Funding

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None

Competing Interests

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No competing interest

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