Reproductive Organ Activities of Morinda lucida Ethanol Root Extract on Male and Female Albino Rats Tramadol HCL Induced Infertility

Infertility is a serious issue disturbing reproductive ages in all society now and calling for solution for continuity. Infertility were induced on groups of animals with separate sexes; M, M1, M2 and M3; F, F1, F2 andF3 with daily subcutaneous administration of 20 mg/kg body weight tramadol HCl for 42 days before respective ethanol root extract administrations of (500, 1000, 1500) mg/ kg body weight for 10 days. The animals were anaesthetized and sacrificed; uterus, ovaries, testes, epididymis were dissected out for histomorphological studies. There is evidence of dose treatment of infertility among treated groups. From the organ weight study, both the male and the female organ weight in the groups remained significantly unchanged. There are dose dependent treatments with the male and female tramadol treatment. The result is more pronounced in the increased dose of ethanol root extract (1000 mg/kg and 1500 mg/ kg) body weight treated testicular Original Research Article Amadi et al.; JPRI, 33(46B): 287-295, 2021; Article no.JPRI.75144 288 cells. This study indicated that Morinda lucida has a prophylactic effect against tramadol-induced testicular damage.


INTRODUCTION
This is a system of sex organ used to produce male and female sex cell; Female reproductive organ comprises of internal structures (cervix, uterus, fallopian tube and ovary) while male sex organ includes; testes, prostrate [1]. The male sex organ is responsible for the production and storage of male gamete: the two testes are an oval shaped organ that produces male gamete and androgen, the during ejaculation prostate gland secrets sperm produced in the testes through ejaculatory duct which carries sperm and the fluid secreted by seminal vesicle to unit area in the urethra [2,3]. The female sex organ ovary prepares and nurture oocyte for ovulation process and release through the uterine tube of fallopian tube into the uterus which nourishes fertile eggs [4].
Drugs interfere with fertility by stimulating the body to impact changes on hormonal balance thereby affecting luteinizing hormone (LH) which secretion causes release of ovaries and follicle stimulating hormone (FSH) by the pituitary gland [5]. Effect on male sex organs involve inhibition in testosterone which is the main hormone that signal testicle in production thus leading to decrease in sperm production [6]. Alcohol consumption and drugs causes infertility, such as steroids, opiates and marijuana affects testosterone and interferes in disruption of ovulation and endometrial reception of sperm cells [7]. Interruption of reactive oxygen species (ROS) level and antioxidant defence system imbalance induced by oxidative stress lead to cellular changes causing damages on these organs [8].

MATERIALS AND METHODS
Studies on activities of ethanol root Morinda lucida extracts on tramadol HCl induced infertility were conducted according to Salah et al. [9] and El-Ghawet, [10] with modifications.

Animals
Sixty (60) adult male and female rats respectively, between 7-9weeks of body weight 120-150g were used in this study.

Crude extraction preparation procedure
The root of the plant was washed, air dried, finely ground into powder using a grinder and wrapped in nylon to avoid contamination. Morinda lucida root extracts were prepared by immersing 250 g of ground powder into to 1000 ml of 80% ethanol respectively before subjection to the following model: The ethanol extractions were left for 72 hrs at room temperature.
1. Resultant crude extractions were obtained by first filtration through the muslin cloth; then further filtrations through Whatman No. 1 filter paper. 2. The filtrates were concentrated using an evaporator set at 40 0 C.
Each jelly concentrate obtained was placed in a well labelled plastic container and stored in a refrigerator at 4 0 C until required.

Experimental Treatment Design
Each sex of albino rats were grouped into 10 groups M, M 1, M 2, M 3 , F, F 1 , F 2 , F 3 , M 0 and F 0 .

RESULTS
Effect on reproductive system following administration of tramadol female reproductive systemovaries.

Relative Organ Weight (ROW) Ovaries and Uterus
There were no significant changes in the relative ovaries and uterus organ weight among the treated groups as shown in Table 1.
The F 0, F 01, F 02 and F 03 group showed normal features in interstitial cells, mature follicles, primary follicles and blood vessels while the F (positive control) group presents normal features with dilated blood vessels. Section of ovary from an animal in Group F 1 shows normal features, but with the blood vessels appearing dilated as shown in slide but F 2 and F 3 group show normal features without cellular alterations.

Uterus
The uteri among all the groups showed normal histoarchitecture without degenerative changes in the epithelia lining, uterine glands and intestinal connective tissue cell.

Relative Organ Weight (Row) of Testes and Epididymis
There were no relative organ weight significant changes observed among the groups as shown in Table 2

DISCUSION
Tramadol is one of the most popular misused opiates used in the treatment of sexual premature ejaculation that can disrupt testosterone production [6,11]. Abuse of tramadol may accumulate into toxic metabolites that increase pharmacokinetic interaction, decrease elimination and induce oxidative stress by decreasing the antioxidant levels in the body thus resulting in toxicity that may accompany infertility [12]. Its effect may involve reduction in luteinizing hormone plasma level (LH), follicle stimulating hormone (FSH), increase in prolactin, estradiol, nitric oxide, lipid peroxidation and decreased antioxidant enzyme activities [5,9].
From the organ weight study, both the male and the female organ weight in the groups remained significantly unchanged. There are dose dependent treatments with the male and female tramadol treatment. The result is more pronounced in the increased dose of ethanol root extract (1000mg/kg and 1500mg/ kg) body weight treated testicular cells. Plants with polyphenols and flavonoids have shown tolerate toxicity effect of tramadol [13]. Furthermore, the phytochemistry of Morinda lucida root extracts revealed a high content of total polyphenols and flavonoids; despite, the seminiferous tubules appear to have severely degenerated germinal epithelium in negative control group by free radical scavenging activity [14].

CONCLUSION
This study indicated that Morinda lucida has a prophylactic effect against tramadol-induced testicular damage.

DISCLAIMER
The products used for this research are commonly and predominantly use products in our area of research and country. There is absolutely no conflict of interest between the authors and producers of the products because we do not intend to use these products as an avenue for any litigation but for the advancement of knowledge. Also, the research was not funded by the producing company rather it was funded by personal efforts of the authors.

CONSENT AND ETHICAL APPROVAL
As per university standard guideline, participant consent and ethical approval have been collected and preserved by the authors