EFFECTS OF Carica papaya MALE FLOWER ON FERTILITY ...

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Erigbali et al. World Journal of Pharmacy and Pharmaceutical Sciences

EFFECTS OF Carica papaya MALE FLOWER ON FERTILITY

PARAMETERS IN SULFASALAZINE INDUCED REPRODUCTIVE

TOXICITY AMONGST MALE ALBINO RATS

*1Peter P. Erigbali,

2Olubunmi O. Ezomoh and

2Sule O. Jimoh

1Department of Human Physiology, Faculty of Basic Medical Sciences, Niger Delta

University.

2Department of Biochemistry, Faculty of Basic Medical Sciences, Niger DeltaUniversity.

ABSTRACT

The effect of carica papaya male flower extract on sulfasalazine

induced reproductive toxicity in male albino rats was studied for a

period of 18 days. The investigation was done with 24 male albino rats,

placed in four (4) groups of six (6) each. Group 1(Normal control) was

given normal saline. Group 2(Positive control) was given normal

saline. Groups 3(Treated group 1) and 4(Treated group 2) were treated

with carica papaya male flower extract once daily at 100mg/kg and

200mg/kg body weight respectively for 18 days. The rats in groups 2,3

and 4 were administered 500mg/kg of sulfasalazine (SASP) once daily

for 3 consecutive days and all the groups (1-4) were allowed free

access to food and water. Serum testosterone, Follicle stimulating hormone (FSH), sperm

count, sperm motility, sperm morphology, sperm pH and histopathology of the testes were

assayed following standard procedure. We observed significantly (p˂0.05) increased weight

gain in all the rats administered SASP, when compared with the normal control (178.57±

17.40). There was significant increase in weight gained by treated group 1(206.17 ± 3.19)

than positive control group (189.85 ± 19.83), but decrease in weight gained by treated group

2 (203.1 ± 11.00) than treated group 1(p˂0.05). The sperm count decreased significantly

(p˂0.05) in groups 2 (76.17 ± 4.63) and 3 (98.17 ± 21.77) compared to 1 (112.5±17.99).

But sperm count increased significantly in groups 3 and 4 (118.17 ± 25.49) compared to 2.

Sperm motility decreased in groups 2 (77.83±11.69), 3(69.45±13.32), 4 (72.33±19.64)

compared to 1 (91.60±4.79) and decreased in 3 and 4 compared to 2. The pH of semen was

slightly increased in groups 2(7.10±0.15), 3(7.10±0.20), 4(7.00±0.54) than 1(6.96±0.08)

WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

SJIF Impact Factor 7.632

Volume 10, Issue 7, 217-233 Research Article ISSN 2278 – 4357

*Corresponding Author

Peter P. Erigbali

Department of Human

Physiology, Faculty of Basic

Medical Sciences, Niger

Delta University.

Article Received on

10 May 2021,

Revised on 01 June 2021,

Accepted on 21 June 2021

DOI: 10.20959/wjpps20217-18999


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statistically (p<0.05), but not different comparing group 2 vs 3 and 4. There was significant

decrease in semen morphology of groups 2(36.5±13.44), 3(37.16±5.57), 4(39.93±4.23)

compared to 1(78.15±9.84), but increase in semen morphology of groups 3 and 4 compared

to 2, although not significant (p<0.05). Result of hormonal profile showed that Testosterone

and FSH were significantly decreased in groups 2(0.55 ± 0.05 and 0.30±0.00), 3(1.2 ± 0.1

and 0.35±0.07), 4(0.85 ± 0.05 and 0.20±0.00) compared to normal control (2.75 ± 0.05 and

0.65±0.07) respectively; also they were significantly lower in group 4 than group 2 (p<0.05).

These observations seem to imply that carica papaya male flower potentially may boost

fertility parameters; including hormonal profile (for FSH, testosterone), sperm count, semen

quality and even repair testicular damage. The extract perhaps has reverse effect on

sulfasalazine induced reproductive toxicity that affects fertility parameters, and thus can be

considered as good treatment regimen for infertility. It also enhanced weight gain in the rats.

KEYWORDS: Fertility parameters, carica papaya, sulfasalazine, infertility.

INTRODUCTION

Since prehistoric times medicinal plants and the flowers have been discovered and used in

traditional medicine practices. These have evolved over the centuries as essential parts of

African civilization and are widely recognized today as representing its rich cultural and

scientific heritage. The increasing demand for medicinal plant products has renewed the drive

for the production of herbal health care formulations, herbal-based cosmetic products, and

herbal nutritional supplements by pharmaceutical industries. Thus, in addition to serving

medical and cultural functions, medicinal plants in Africa have economic importance. Global

and national markets have been growing for medicinal herbs, and significant economic gains

are being realized through the sale of medicinal plant products (Dzoyem, 2013).

Recently, it was estimated that 80 percent of people worldwide rely on herbal medicines for

some aspect of their primary health care needs, and about 21,000 plant species have the

potential to be used as medicinal plants. Meanwhile, over three-quarters of the world

population rely mainly on plants and plant extracts for their health care needs; with

projected spread that in developed countries such as United States, plant drugs constitute as

much as 25% of the total drugs, while in fast developing countries such as India and China,

the contribution is as much as 80%. (Bodeker et al, 2005; Bandaranayake, 2006).

The male carica papaya flower has been traditionally used as herbal medicine by ancient


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people for centuries in treating different kinds of illnesses. It contains medicinal properties

that are useful for the body in resolving specific health issues including prevention of cancer

and heart disease, reduction of blood sugar levels and promoting the production of insulin

thereby curing diabetes (Nelz 2017). According to Setu, (2019) the extract from the flower,

when mixed with honey is used to treat respiratory problems such as cough and hoarseness of

throat. This flower is rich in folate and antioxidant, giving it potential to fight and reduce bad

cholesterol levels. However, there is not much scientific literature whether this herb is

beneficial in reproductive health.

Reproductive health is the condition of male and female reproductive systems during all life

stages. The systems are made of organs which include ovaries in female and testes in male.

The ovaries, testes produce and release hormones as well as maintain reproductive health in

their respective systems.

For instance, testosterone is a hormone produced primarily by leydig cells inside the testes

and secreted mainly by the testes of males but to a lesser extent, the ovaries of females. In

male humans, testosterone plays a key role in the development of male reproductive tissues

such as testes and prostate, as well as promoting secondary sexual characteristics such as

increased muscle and bone mass, and the growth of body hair. Testosterone helps in the

development of mature sperm and as such, inadequate amount of the hormone will affect the

production of sperm which can lead to infertility in males.

Follicle stimulating hormone (FSH) is one of the gonadotrophic hormones, the other being

the Luteinising hormone. Both are released by the pituitary gland into the bloodstream. FSH

is one of the hormones essential to pubertal development and men’s testes. The successful

and complete male germ cell development is dependent on the balanced endocrine interplay

of hypothalamus, pituitary and the testis. Gonadotropin releasing hormone (Gnrh) secreted by

the hypothalamus elicits the release of gonadotropins i.e. follicle stimulating hormone (FSH)

and luteinizing hormone (LH) from the pituitary gland (De Krester, 1979). FSH binds with

receptors in the sertoli cells and stimulates spermatogenesis. LH stimulates the production of

testosterone in Leydig cells, which in turn may act on the Sertoli and peritubular cells of the

seminiferous tubules and stimulates spermatogenesis (O’Donnel et al., 1994). The failure of

pituitary to secret FSH and LH will result in disruption of testicular function leading to

infertility. Testosterone, estradiol and inhibin control the secretion of gonadotropins

(Weinbauer and Nieschiag, 1995). An increased FSH level in men with azoospermia or


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severe oligozoospermia (<5million sperm/ml) indicate damaged seminiferous tubule

(Bergmann et al., 1994). Studies have shown that in infertile men with germinal epithelial

injury, no stimulation of spermatogenesis occurs due to low or lack of production of

androgen binding proteins. Also the production of inhibin and testosterone,

dihydrotestosterone and estradiol is affected causing disturbance in negative feedback

mechanism resulting in elevated levels of FSH (Gnessi et al., 1997). Although studies have

been carried out on the role of FSH in infertile males, very few studies were carried out on

hormonal levels in different subgroups of infertile males.

Sperm motility is the measure of the percentage of sperm that are in motion and the

percentage of sperm with progressive flagella motion. It is characterized by the movement of

spermatozoa in a fresh sperm. The motion can be divided into four different grades:

Grade a: Sperm with progressive motility. These are the strongest and swim fast in a straight

line. Sometimes it is also denoted motility IV.

Grade b: (non-linear motility): These also move forward but tend to travel in a curved or

crooked motion. Sometimes also denoted motility III.

Grade c: These have non-progressive motility because they do not move forward despite the

fact that they move their tails. Sometimes also denoted motility II.

Grade d: These are immotile and fail to move at all. Sometimes also denoted motility I.

Sperm count is the concentration of sperm per ejaculation or per measured semen. According

to WHO (1992), over 15 million sperm per milliliter is considered normal.

Sperm morphology refers to the size and shape of individual sperm. It is one of the

contributing factor to male fertility. Sperm morphology results are reported as the percentage

of sperm that appears normal when semen is viewed under a microscope.

The pH of semen is determined by acidic secretions of the prostate and alkaline secretions of

the seminal vesicles. It should normally be in the range of 7.2-8.0 (Cooper et al., 2010). If the

pH exceeds 8.0, infection should be suspected with decreased secretion of acidic products by

the prostate, such as citric acids. Abnormal pH may also be recorded in cases of incomplete

ejaculation. Extremely acidic pH (<6.5) is found in the cases of agenesis (or occlusion) of the

seminal vesicles.

The human testes are two organs of the shape of rotation ellipsoids with diameters of 2.5 -


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4 cm engulfed by capsule (tunica albuginea) of strong connective tissue (Davidorff et al.,

2002). Thin septula testis divide the parenchyma of the testis in about 370 conical lobules.

The lobules consist of the seminiferous tubules and intertubular tissue, containing groups of

endocrine leydig cells and additional cellular elements. The fluid secreted by the

seminiferous tubules is collected in the rete testis and delivered to the ductal system of the

epididymis.

The testes tubules are surrounded by connective tissues stromal cells which contain

testosterone secreting leydig (interstitial) cells. The tubules are lined with a layer of

seminiferous epithelium, which contains supporting sertoli (sustentacular) cells. The

spermatogenic cells constantly multiply and through several phases of spermatogenesis

differentiate into mature sperm, while the sertoli cells nourish them.

Reproductive toxicity has become a problem of great interest and concern in recent times.

Humans are exposed to toxic substances on a daily basis from air, water, food and drugs that

are taken to the body. Some of these drugs have toxic effect on some organs or tissues of the

body and may cause serious health problems. Some of the toxic effects of these substances

affect the reproductive system of humans thereby causing induced – infertility problems. In

recent times, there is an increasing number of infertility problems that affect males which can

be from the result of various drugs. Some of these drugs are antibiotics (Sulfasalazine),

antidepressants or thalidomide (Wikipedia, 2019).

Sulfasalazine is a drug commonly used for the treatment of rheumatoid arthritis, ulcerative

colitis, Crohn’s disease (Wikipedia, 2019) and Inflammatory bowel disease (Rowe, 2017).

Sulfasalazine, a drug used in the treatment of inflammatory bowel disease, has been

associated with male infertility, an effect attributed to sulfapyridine rather than to 5-

aminosalicylic acid (5-ASA), the presumed therapeutically active component of sulfasalazine

(Chatzinoff, et al; 1988). It has been confirmed by various researchers that sulfasalazine

causes temporary infertility and that prolong treatment with sulfasalazine may universally

depress quality of semen and cause reversible infertility (Toovey et al., 1981). The results of

previous studies by Fukushima, et al (2005), hypothesize that the suppression of epididymal

acrosomal membrane proteins synthesis with their consequent reduced incorporation to the

sperm membrane leads to a depressed sperm motility and acrosome reaction, and thereby

leads to infertility in SASP treated male rats. Taking the foregoing available information into

account and the fact that not much seems to be reported, on any scientific association


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between carica papaya male flower and fertility; the aim of this research therefore was to

study the effect of carica papaya male flower on sulfasalazine – induced infertility in wistar

albino rats. And we specifically investigated sperm motility, sperm morphology, sperm

count, semen pH, FSH, testosterone and histopathology of the testes.

METHODOLOGY

Apparatus/Materials

The apparatus and materials used in the experiment were: spectrophotometer (Medifield

Equpment & Scientific LTD.ENGLAND), dissecting set, centrifuge, water bath, oven (Bio-

tech), weighing balance, spatula, test tubes, refrigerator, plain container, microscope, cotton

wool, grease free slide, neubauer counting chamber, cover slip, syringe, stirring rod, electrical

blender and orogastic tube (gavage).

Chemical and reagents Sulphasalazine, chloroform, 10% formalin, ethanol, normal saline,

carica papaya male flower extract, urea test kits (Randox) and Creatinine test kits.

Experimental animals

Twenty-four (24) male albino rats weighing 100- 180g were used in the study. The rats were

gotten from the animal house in the Department of biochemistry, university of Port Harcourt,

Rivers State, Nigeria. They were kept in a standard cage in the animal house of Department

of pharmacology, Niger Delta University, Wilberforce Island, Bayelsa state, Nigeria and

acclimatized for three weeks during which they were fed daily with food (pelleted grower

crumbs) and distilled water.

Sources of Sample

Carica papaya male flower

Fresh flowers from male carica papaya were gotten from different locations in Amassoma,

Sourthern Ijaw local Government Area, Bayelsa State, Nigeria. The plant was identified

with the voucher number FA/HERB/VN/008C at the Department of Agriculture, Niger Delta

University Wilberforce Island, Bayelsa State Nigeria.

Tablet (Sulphasalazine)

The tablet was gotten from a reputable pharmacy.


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Preparation of sample

Flower extract

The fresh flowers were cleaned of debris and dirt and oven dried for three (3) days. The dried

flowers were homogenized to a powdered form with the help of an electrical blender. 292g

of the powder was placed in a jar with 2 litres (2L) of ethanol. The mixture was stirred

vigorously and allowed to stand for 24 hours (1 day) with frequent stirring. The mixture was

strained and the flower extract was concentrated in the oven at 50 – 55oc for 1 week (7 days)

to yield black syrup.

Sulfasalazine

1500mg of sulfasalazine tablet were mashed in a mortar and transferred to a beaker where it

was mixed with 50mL of normal saline and stored in the refrigerator for use.

Experimental Design

GROUP 1 (normal control group): Food + Normal saline GROUP 2 (Positive control Group):

Food + Normal saline.

GROUP 3 (Treated group 1): Food + 100mg of flower extract daily. GROUP 4 (Treated group

2): Food + 200mg of flower extract daily.

The rats in groups 2,3 and 4 were administered 500mg/kg body weight of sulfasalazine once

daily for 3 consecutive days and all the groups (1-4) were allowed free access to food and

water.

Body Weight

At the end of the administration of extract for eighteen (18) days, the body weight of each

rat was recorded.

Collection of Blood Sample

The animals were treated with chloroform to induce unconsciousness. They were dissected

and blood was collected directly from their heart with a 5mL syringe into and plain sample

bottle. The blood was allowed to coagulate and centrifuged at 3000rpm for 10 minutes. The

serum was collected and used for hormonal assays.

Semen Collection

After collection of blood, the testes were removed and the epididymis was isolated and

placed in a petri containing 0.5mL of normal saline at 37oC. 0.5mL of the sperm suspension


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was added to 1.5ml of normal saline and subsequently mixed gently.

Testes Collection

The right and left testes were collected and weighed on a weighing balance, after which they

were preserved in formal saline for further experiments.

Determination of Hormonal Profile

Testosterone

Serum from each sample was assayed for testosterone using the Enzyme-linked

immunosorbent assay (ELISA) techniques using the Accu Bind ELISA microwells

Testosterone test system for testosterone and in accordance with standard protocols

outlined by the kits producer.

Follicle Stimulating Hormone (FSH)

The concentration of follicle stimulating hormone (FSH) was determined using the enzyme

linked immunosorbent assay technique (ELISA) using the Accu Bind ELISA microwells

Follicle Stimulating Hormone (FSH) test system for FSH and in accordance with standard

protocols outlined by the kits producer.

Semen Analysis.

Sperm Motility

A drop of semen was placed on a clean grease – free slide attached to a hemocytometer

chamber and was examined under the microscope using a magnification of X10. The total

number of motile, sluggish or dead spermatozoa was noted. The percentage of motile sperm

was gotten by dividing the number of motile sperm by the total number of sperm counted per

box.

Sperm Count

One drop of sperm suspension was loaded into hemocytometer chamber and the sperm in the

squares (5 diagonal squares) of the hemocytometer were counted under the microscope X10.

The count was calculated as the number of spermatozoa per mL epididymal suspension.

Calculation


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Semen pH

The sperms were collected from the epididymis into a Petri dish containing 1ml of normal

saline with pH 6.0 for group one and two and 1.5ml of normal saline for group three and four

to make a semen suspension after which a pH meter was used to take the pH of the semen

from each sample.

Sperm Morphology

The sperms were collected from the epididymis into a Petri dish containing 1ml of normal

saline with pH 6.0 for group one and two and 1.5ml of normal saline for group three and four

to make a semen suspension after which a drop of semen was put on a grease free slide and

placed under an electron microscope for reading using x10 magnification.

Histopathological Examination

The rats were sacrificed at the end of experiment, testes removed and fixed in chloroform for

histological studies. The method of Baker and Silverton (1985) was adopted in the

preparation of slices of the fixed testes for histological examination.

RESULTS

Table 1: Effect of Sulfasalazine and carica papaya male flower on the weight of albino

rats.

Weight of albino rats (g)

Group 1 (Normal Control) 178.57± 17.40d

Group 2 (Positive Control) 189.85 ± 19.83c

Group 3 (treated group 1 (100mg/kg extract) 206.17 ± 3.19a

Group 4 (treated group 2 (200mg/kg extract) 203.1 ± 11.00b

Values are recorded as MEAN±SD (n=6). Means with different superscript letters are

statistically different at 95% confidence level (p≤0.05).

The weight of animals in all the groups was recorded and the result showed that there was a

significant (p≤ 0.05) increase in the body weight of the test groups compared to both control

groups. Also, significantly increased weight in positive control than the normal control group

was observed.


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Hormonal profile

Table 2: Effect of sulfasalazine and carica papaya male flower on hormonal profile.

Testosteron e

(ng/mL) FSH (mIU/ml)

Group 1 (Normal Control) 2.75 ± 0.05a 0.65±0.07

a

Group 2 (Positive Control) 0.55 ± 0.05c 0.30±0.00

b

Group 3 (test group with 100mg/kg extract) 1.2 ± 0.1b 0.35±0.07

b

Group 4 (test group with 200mg/kg extract) 0.85 ± 0.05c 0.20±0.00c


statistically different at 95% confidence level (p≤0.05). Hormonal profiling indicates a

significant (p≤0.05) decrease of testosterone level in groups 2(0.55 ± 0.05), 3(1.2 ± 0.1) and

4(0.85 ± 0.05) rats administered with sulfasalazine when compared to normal control

(2.75±0.05). However, there was a significant (p≤0.05) increase of testosterone level in group

3(1.2±0.1) than group 2, and a non-significant (p≤0.05) increase of same in group

4(0.85±0.05) than 2. There was a significant (p≤0.05) decrease of FSH in groups 2

(0.30±0.00), 3(0.35±0.07) and 4(0.20±0.00) compared to group 1(0.65±0.07). Also, a

significant (p≤0.05) decrease of FSH in group 4 than 2, and a non-significant increase of

same in 3 than 2.

Semen Analysis

Table 3: Effect of sulfasalazine and carica papaya male flower on semen quality.

Sperm Count

Nox106sperm/M l

Motility (%) Semen

pH

Morphology

(%)

Group 1 (Normal Control) 112.5±17.99a 91.60±4.79

a 6.96±0.08

a 78.15±9.84

a

Group 2 (Positive Control) 76.17 ± 4.63c 77.83±11.6 9

b 7.10±0.15

b 36.5±13.44

b

Group 3 (test group with

100mg/kg extract) 98.17 ± 21.77

b 69.45±13.3 2

c 7.10±0.20

b 37.16±5.57

b


statistically different at 95% confidence level (p≤0.05) for these parameters. A significant

(p≤0.05) decrease in the number of sperm cell was observed in group 2, (76.17 ± 4.63) and

3(98.17 ± 21.77) but non-significant increase in group 4(118.17 ± 25.49), compared to

1(112.5±17.99). However, the number of sperm cells increased significantly (p≤0.05) in

groups 3 and 4 that had the extract of carica papaya than 2 which never had the extract.

Sperm motility was significantly (p≤0.05) decreased in groups 2(77.83±11.69),

3(69.45±13.32), 4(72.33±19.64) compared to 1(91.60±4.79).

Also, motility was reduced in groups 3 and 4 than 2. Semen pH was slightly increased

significantly in group 2,3,4 compared to 1(p≤0.05), but no significant difference was


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observed among 2,3 and 4. Meanwhile, sperm morphology was significantly decreased in

groups 2,3,4 compared to 1(p≤0.05), but no significant difference was observed among 2,3

and 4; rather the observed increase in morphology of groups 3 and 4 over 2 was not

significant at (p<0.05).

Weight of Testes

Table 4: Effect of sulfasalazine and carica papaya male flower on weight of testes.

Weight of testes (g)

Group 1 (Normal Control) 3.03 ± 0.29a

Group 2 (Positive Control) 2.93 ± 0.41b

Group 3 (test group with 100mg/kg extract) 3.18 ± 0.29a

Group 4 (test group with 200mg/kg extract) 2.90 + 0.16b


statistically different at 95% confidence level (p≤0.05)

The weight of testes recorded for all groups showed that it was significantly decreased

(p<0.05) in group 2 and 4 than 1. But non-significantly increased in group 3 than 1.

However, it was significantly increased in group 3 that had extract of papaya than group 2

that never had the extract. And the observed decreased weight of testes in group 4 compared

to 2 was non-significant.

Histopathology

Fig. 1: Photomicrograph of testicular section of group 1 administered with normal

saline showing normal architecture.

Microscopic examination of testicular sections of this control group revealed rounded

seminiferous tubules with regular outlines. The cells of the germinal epithelium at different

stages of spermatogenesis. The flagella of mature sperms were seen filling the lumens of the


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seminiferous tubules. The interstitial spaces in-between the tubules contained interstitial

cells of Leydig. The germinal epithelium consisted of spermatogonia cells resting on the

basement membrane, large rounded primary spermatocytes with large vesicular nuclei; also

spermatid could be recognized by its darkly stained rounded nuclei and its position towards

the lumen, while spermatozoa appeared elongated. Conclusion: Consistent with normal

histology of the testis.

Fig. 2: Photomicrograph of testicular section of group 2 stained with haematoxylin and

eosin that was administered with normal saline and sulfasalazine (500mg/kg) showing

interstitial necrosis and spermatogonia with altered architecture.

Microscopic examination of testicular sections in group 2 revealed germinal epithelium at

different stages of spermatogenesis with increase sperm cells production. The cell shows

cellular hypertrophy. Also seen is extensive interstitial necrosis within the interstial space in

between the tubules. Conclusion: Inconsistent with normal histology of the testis

Fig. 3: Photomicrograph of testicular section of group 3 stained with haematoxylin and

eosin that was administered with normal saline, sulfasalazine (500mg/kg) and carica

papaya male flower extract (100mg/kg) showing interstitial necrosis and with altered

architecture.


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Microscopic examination of testicular sections of group revealed germinal epithelium at

different stages of spermatogenesis. The spermatogonia are seen detached from the basement

membrane. Micrograph shows altered histomorphology of the interstitial spaces in-between

the tubules with various degree of necrosis and exudation Conclusion: Inconsistent with

normal histology of the testis

Fig. 4: Photomicrograph of testicular section of group 4 stained with haematoxylin

and eosin that was administered with normal saline, sulfasalazine (500mg/kg) and

carica papaya male flower extract (200mg/kg) showing normal seminiferous tubules.

Microscopic examination of testicular sections of group 4 revealed normal histology of

the testicular as seen in group 1. Conclusion: Consistent with normal histology of the

testis.

DISCUSSION

The relevance of medicinal flowers in modern world medication cannot be over emphasized,

as parts of the world today depend on traditional system of treatment (Hao, 2019). Carica

papaya male flower has been investigated and found to have many phytochemicals such

as flavonoids, sterols, triterpenoids, tannins, polyphenols and cardiac glycosides, with several

medicinal properties (Bergonio & Perez, 2015). This report was corroborated by a

preliminary phytochemical studies carried out by Udurume in 2019 (unpublished article).

In this research, we specifically studied effect of carica papaya male flower on sulfasalazine

– induced infertility in wistar albino rats by investigating sperm motility, sperm morphology,

sperm count, semen pH, FSH, testosterone and histopathology of the testes in the

experimental model. The results showed that testosterone level significantly decreased in

groups 2,3,4 that had sulfasalazine than group 1(normal control) that was not administered

sulfasalazine. But there was significant increase in testosterone level of group 3 compared to


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group 2, and a non-significant increase in group 4 compared to group 2 which all suggests

that sulfasalazine had activity that resulted in reduced testosterone levels associated with

infertility for rats in groups 2,3 and 4, but extracts of carica papaya male flower had activity

that alleviated this effect for rats in group 3(with statistical significance) and 4. The increase

in group 4 was not statistically significant implying that this activity may not be dose-

dependent. Also, Sperm count was significantly reduced in groups 2 and 3 that were

administered sulfasalazine compared to 1 (normal control) that did not receive it. But there

was significant increase in sperm count of rats in group 3 and 4 that were given extracts of

carica papaya flower than group 2(positive control). And these imply that sulfasalazine’s

infertility related activity was attenuated in the rats that were administered extracts of carica

papaya. Although carica papaya was reported to have contraceptive capability, besides its

digestive, antibacterial, anti-inflammatory, anti-ageing, and other effects on health (Madinah,

Nozmo & Iliya, 2015), this finding did not corroborate contraceptive potential of papaya male

flower as reported.

The observation seems to agree with earlier reports that testosterone regulates sexual

development, growth, pubertal maturation and reproductive qualities by enhancing the

induction and maintenance of normal sperm production (Vasudevan et al., 2011; Simoni et

al., 1999).

We observed a non-significant increase of FSH level in group 3 than 2, but significant

decrease of FSH in group 4 than 2. Relating this with the histopathologic investigations; it

appears lower dose of papaya extract may have no observable effect, but in higher dose it

reduced the FSH level which was increased perhaps due to damage caused by sulfasalazine as

seen in the altered histomorphology. This and the preceding discussions can corroborate

report that FSH level within normal range stimulates the growth and maturation of sperm cell

(Furman and John, 2015; Wanda, et al; 2010).

There was significant (p<0.05) increase in sperm morphology of the rats and the number of

sperm cells in groups 3 and 4 compared to the positive control group (2), implying that carica

papaya attenuated effect of sulfasalazine and or improve fertility parameter. It is conceivable

that, since carica papaya male flower extract is rich in triterpenoids and flavonoids which are

good antioxidants, these may have contributed in neutralizing the effect of free radical

reactive oxidative species caused by sulfasalazine; corroborating the research carried out by

Panche et al (2016) on flavonoids and its antioxidant activity. The extract also appears to


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have attenuated the effect of sulfasalazine on motility of the sperm but this was not significant

at p˂0.05.

As shown in the histopathological study, sulfasalazine caused interstitial necrosis in the testes

in the positive control group. This effect was reversed in group 4 administered with a higher

concentration of the extract. This suggests that the extract may be capable of repairing

testicular damage. From the results it may be inferred that carica papaya male flower has

potential to curb reproductive toxicity induced by sulfasalazine. Perhaps this is due to its

rich phytochemical constituent that include antioxidants; thus it was able to prevent free

radical injury and facilitate repair of damaged tissues (Malik, et.al., 2017).

CONCLUSION

In this study, the observations suggest that carica papaya male flower may potentially boost

fertility parameters; such as relevant hormonal profile, sperm count, semen quality and even

repair testicular damage. The extract perhaps has reverse effect on sulfasalazine induced

reproductive toxicity that affects fertility parameters, and therefore may serve as a good

treatment regimen for infertility. However, further research is required to identify the active

ingredient(s) in carica papaya male flower and elucidate the exact mechanism of action.

REFERENCES

1. Dzoyem, J.P., Tshikalange E. & Kuete V. Medicinal Plants Market and Industry in

Africa. Pharmacology and Chemistry, First edition, CHP., 2013; 24: 859–890. Elselvier.

2. Bodeker C., Bodeker G., Ong C.K., Grundy C.K., Burford G., Shein K. WHO Global

Atlas of Traditional, Complementary and Alternative Medicine. Geneva, Switzerland:

World Health Organization, 2005.

3. Bandaranayake W.M. ‘’Quality Control, screening, toxicity and regulation of herbal

drugs,’’ in Modern Phytomedicine. Turning Medicinal plants into drugs. Chp., 2006; 2:

25 – 57.

4. Nelz, J., (2017), Discover the Health Benefits of Papaya Flower. Retrieved March 2017.

From. www.philnews.ph

5. Setu, J. (2019), Papaya Flowers and Leaves Benefits. Retrieved 2019. From

www.jaiviksetu.com

6. De Krester, D.M. Endocrinology of Male Infertility. Brit. Med. Bullet, 1979; 35(2): 187-

192. Department of Health Office of the gene technology regulation, (2008), The

Biology of Carica papaya L. Retrieved February 2008. From www.ogtr.gov.au

http://www.philnews.ph/

http://www.jaiviksetu.com/

http://www.ogtr.gov.au/


232


7. O’Donnel, L., R.I., Mc Lachlan, N.G., Wreford and Robertson, D.M. Testosterone

promotes the conversion of round spermatids between stages vii and viii of the rat

spermatogenic cycle. Endocrinology, 1994; 135(4): 2608-2614.

8. Weinbauer, G.F. and Nieschlag, E. Gonadotropin control of testicular germ cell

development. Adv. Exp. Med. Biol., 1995; 317: 55-65.

9. Bergmann, M., Behre, H. and Nieschlag, E. Serum FSH and testicular morphology in

male infertility. Clin. Endocrinol, 1994; 40: 133-136.

10. Gnessi, L., Fabbri, A. and Spera, G. Gonadal peptides as mediators of development and

functional control of the testis. An integrated system with hormones and local

environment, 1997; 18: 541.

11. WHO. World Health Organization Laboratory Manual, 1992.

12. Cooper T.G., Noonan E., Ekardstein S.V., Auger J., Baker G.H.W. Human Reproduction

Update, NIH., 2010; 16(3): 231–245.

13. Davidorff M.S., Middendorff R., Kofuncu E., Muller D., Jesek D & Holstein A.F. Leydig

cells of the human testis possess astrocyte and oligodendrocyte marker molecules. Acta

Histochem, 2002; 104(1): 39–49.

14. Wikipedia (2019), Reproductive Toxicity Retrieved. September 2019. From

www.en.m.wikipedia.org

15. Rowe, W. A. (2017), How Does Sulfasalazine Affect the Fertility of Men with

Inflammatory Bowel Disease (IBD)? Retrieved October 2017. From www.medscape.com

16. Chatzinoff, M. & et.al, Sulfasalazine – induced Abnormal Sperm Penetration Assay

Reversed on Changing to 5-aminosalicyclic acid enemas. Digestive Diseases and

Sciences, 1988; 33(1): 108–110.

17. Toovey, S. & et.aln (1981), Sulfasalazine and Mlae Infertility Reversibility and Possible

Mechanism. Retrieved 1981. From www.ncbi.nilm.nih.gov

18. FukushimaT., Kato M., Adachi T., Hamada Y., Horimoto M., et al. Effects of

Sulfasalazine on Sperm Acrosome Reaction and Gene Expression in the Male

Reproductive Organs of Rats. Toxicological Sciences, 2005; 85(1): 675–682.

19. Baker, F. J. & Silverton, R. E., Medical Laboratory Technology; Diagnosis, Laboratory;

Technology. Medical Butterworths (London and Boston) 6th

Edition, 1985; 408.

20. Hao, D. (2019), Genomics and Evolution of Medicinal Plants Retrieved 2019. From

www.sciencedirect.com

21. Bergonio, K. B. & Perez, M. A., The Potential of Male Papaya (Carica papaya, L.)

Flower as a Functional Ingredient for Herbal Tea Production. Indian Journal of

http://www.en.m.wikipedia.org/

http://www.medscape.com/

http://www.ncbi.nilm.nih.gov/

http://www.sciencedirect.com/


233


Traditional Knowledge, 2016; 15(1): 41–49.

22. Udurume, 2019 (Unpublished article)

23. Madinah, N. et al, The Protective Effects of A queos extract of Carica papaya seeds in

paracetamol induced nephrotocity in male wistar rats. African Health Sciences, 2005;

15(2): 598-605.

24. Vasude van D.M., Sreekumari S., Kannan V. Steroid hormones; Testicular hormones.

Text Book for Medical students, 6th

ed. JAPEE brothers medical publishers LTD., 2011;

532–537.

25. Simoni M., Weinbauer GF., Gromoll J., Nieschlag E. Role of FSH in gonadal function.

Ann Endocrinol, 1999; 60(2): 102–106.

26. Furman, B. L. & John, S. (2015), Follicle Stimulating Hormone. Reference Module in

Biomedical Sciences. www.sciencedirect.com

27. Wanda, M. H. et al; (2010), Male Reproductive System. Fundamentals of Toxicologic

Pathology 2nd

Edition

28. Panche A.N., Diwan A.D., & Chandra S.R. Flavonoids; an overview. Nutrition

Science, 2016; 5: 41-47.

29. Malik, A. et.al, Male Infertility: The Effect of Natural Antioxidants and Phytocompounds

on Seminal Oxidative Stress. Diseases, 2017; 5(1): 9. doi:10.3390/diseases5010009

http://www/

EFFECTS OF Carica papaya MALE FLOWER ON FERTILITY ...

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