







































_____________________________________________________________________________________________________ 
 
*Corresponding author: E-mail: mfonakpaso@unical.edu.ng; 
 
Cite as: Akpaso, Mfon I, Elton N Takim, Michael E Oku, Anani Sadeyeng, Lydia B Edim, and Gabriel U Udo-Affah. 2024. 
“Gonadal Histo-Morphology and Enhanced Fertility Potential of Curcuma Longa in Male STZ-Induced Diabetic Rats”. Asian 
Journal of Immunology 7 (1):258-73. https://doi.org/10.9734/aji/2024/v7i1149. 
 

 
 

Asian Journal of Immunology 
 
Volume 7, Issue 1, Page 258-273, 2024; Article no.AJI.126504 
 

 
 

 

 

Gonadal Histo-morphology and 
Enhanced Fertility Potential  

of Curcuma longa in Male  
STZ-induced Diabetic Rats 

 
Mfon I Akpaso a*, Elton N Takim a, Michael E Oku a,  

Anani Sadeyeng a, Lydia B Edim a and Gabriel U Udo-Affah a 
 

a Department of Anatomy, Faculty of Basic Medical Sciences, University of Calabar, Calabar, Nigeria. 
 

Authors’ contributions  
 

This work was carried out in collaboration among all authors. All authors read and approved the final 
manuscript. 

 
Article Information 

 
DOI: https://doi.org/10.9734/aji/2024/v7i1149 

 
Open Peer Review History: 

This journal follows the Advanced Open Peer Review policy. Identity of the Reviewers, Editor(s) and additional Reviewers,  
peer review comments, different versions of the manuscript, comments of the editors, etc are available here: 

https://www.sdiarticle5.com/review-history/126504 

 
 

Received: 14/09/2024 
Accepted: 18/11/2024 
Published: 04/12/2024 

 
 

ABSTRACT 
 

Diabetes mellitus is a disorder in which the body fails to produce enough or respond normally to 
insulin, leading to abnormally high blood sugar levels. The study was aimed at investigating the 
effects of ethanolic extract of Curcuma longa (Tumeric) on fertility profile and gonadal histo-
morphology of streptozotocin-induced hyperglycemic male Wistar rats. Semen parameters (count, 
motility and viability) and testicular histology of streptozotocin “STZ”-induced hyperglycemic male 
Wistar rats were investigated following the administration of Curcuma longa rhizome extract. 
Twenty-four (24) rats weighing 110-180g were grouped into four. The groups were labelled A to D 
and contained six rats each. Group A (normal control) rats were given rat pellets and distilled water 

Original Research Article 

https://doi.org/10.9734/aji/2024/v7i1149
https://www.sdiarticle5.com/review-history/126504


 
 
 
 

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259 

 

only. Group B (diabetic control) rats were given 65mg/kg body weight “BW” of streptozotocin IP. 
Group C served as the standard group and was given 65mg/kg of streptozotocin IP + 250mg/kg of 
metformin IP, while Group D (test group) rats were given 65mg/kg BW of streptozotocin IP + 
500mg/kg of Curcuma longa extract. Curcuma longa extract was administered through the oral 
route with the aid of an oro-gastric tube. The administration lasted for twenty-eight (28) days. The 
rats were then anaesthetized and the testes and semen obtained for histological and semen 
analysis (sperm count, sperm motility, sperm viability) respectively. The diabetic group showed 
seminiferous tubules with reduced germinal epithelium and distorted interstitial connective tissue 
with a significant (p<0.05) reduction in sperm motility, count, viability compared to the normal group 
that showed normal histological features and normal sperm parameters (motility, count and 
viability). However, the administration of C. longa showed significant (p<0.05) increase in levels of 
sperm parameters (motility, count and viability) and progressive restoration of histological integrity 
within the testes.  
In conclusion, ethanolic extract of C. longa has the potential to reverse the damage caused by 
hyperglycemia in male reproductive function. 

 

 
Keywords: Curcuma longa; semen analysis; testis; diabetes mellitus. 
 

1. INTRODUCTION 
 
Diabetes mellitus “DM” is a metabolic disorder 
with a characteristic high level of blood sugar 
(hyperglycemia) over a prolonged period of time 
and is usually accompanied by disturbed 
metabolism of proteins and fats [1]. Blood 
glucose rises because it cannot be metabolized 
in the cells due to lack of insulin production by 
the pancreas, or the inability of the cells to 
effectively use the insulin that is being produced 
[1]. Insulin is produced by the beta cells of the 
pancreas and its function is to regulate the 
uptake of glucose from blood into the different 
cells and tissues [2]. Three major symptoms of 
Diabetes mellitus include frequent urination 
(Polyuria), increased hunger and appetite 
(polyphagia) and increased thirst (polydipsia) [1]. 
DM is also associated with complications which 
include diabetic ketoacidosis, cardiovascular 
disease, foot ulcers, damage to nerves and eyes, 
encephalopathy, cardiomyopathy [1,3,4] and 
reproductive dysfunction [5].  
 
The diabetic complications on different cells, 
tissues and organs have been linked to the 
reactive oxygen species (ROS) that are 
generated due to hyperglycemia [6]. Prolonged 
and poorly controlled diabetes has been reported 
to cause sexual dysfunction in males and 
females. Sexual dysfunction can also be an early 
sign of diabetes [7]. In males, diabetes mellitus 
has effects on the endocrine control of 
spermatogenesis and causes erectile, as well as 
dysfunction and impaired ejaculation [8]. It also 
negatively impacts sperm parameters such as 
volume, count, motility and morphology [9]. Due 
to the above-mentioned complications, it became 

imperative by scientists around the world to find 
affordable solutions. Studies have shown that to 
achieve a good metabolic control of diabetes and 
energy balance, a combination of lifestyle, 
nutrition, exercise and pharmaceutical treatment 
are important [10]. They are also said to be major 
factors in the management of diabetes [11]. 
 
Pharmaceutically, Metformin is considered to be 
the first-choice agent for treatment of diabetes 
[12]. Another class of drugs of choice is 
Sulfonylureas [13]. These drugs however are 
seen to have undesirable side effects. On 
account of these side effects, there is advocacy 
for the use of medicinal plants [14], because they 
have been found to have little or no side effects 
and are easily affordable and affordable. The use 
of medicinal plants as an alternative medicine 
has in the last millennium, been accepted all over 
the world [15]. In the United States of America for 
instance, about 38% of the population uses 
herbal medicine [16] and in Turkey, 48.8% of the 
population are said to use herbal medicines [17] 
to treat chronic diseases and illnesses. In Africa, 
the World Health Organization states that at least 
80% of the population relies on medicinal plants 
[18]. Some of these plants and herbs possess 
antioxidant properties. They contain carotenoids, 
flavonoids, alkaloids, glycosides and some are 
said to have anti diabetic effects [19]. One of 
such plants is Curcuma longa commonly called 
turmeric. 
 
Curcuma longa, the turmeric plant is commonly 
used as a spice but has been recognized by the 
scientific community for its antioxidant property. 
In Asia, it has been used as a natural therapeutic 
medicine since ancient times [20]. Its main 



 
 
 
 

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constituent is curcuminoid which gives it a wide 
range of pharmacological properties including 
antioxidant, anti-protozoan, anti-venom, anti-
inflammatory [21], antibacterial, antidiabetic, 
antiviral and anticancer activities [19]. It would 
therefore be interesting to evaluate its 
antidiabetic effects and its effects on the fertility 
profile of streptozotocin-induced hyperglycemic 
male Wistar rats. 
 
Infertility affects about 8-12% of couples 
worldwide and an estimated 50-80 million 
couples have been reported to suffer from some 
form of infertility [22]. Previously, infertility was 
thought to be a problem of the females but 
studies have now shown that 20-30% of infertility 
is linked to the males [23] commonly due to poor 
semen quality or quantity [24]. In 2015 it was 
observed in a study that type 1 diabetic patients 
showed a lower level of spermatozoa with 
increased motility, altered mitochondrial function 
and post ejaculatory dysfunction of the 
epididymis [8]. Diabetes has also shown an 
increase in prevalence over the years with an 
estimated 425 million persons worldwide and 
nearly 50% of that number is not diagnosed [25]. 
Studies have shown that there is a decline in 
fertility involving males with diabetes, leaving a 
high percentage of them with reproductive 
dysfunctions including reduced libido and 
impotence [5]. 
 
STZ is an alkylating agent that causes pancreatic 
islet β-cell destruction and is widely used 
experimentally to produce a model of type 1 
diabetes mellitus (T1DM) [26]. It is a cytotoxic 
glucose analogue which has been used as a 
chemotherapeutic agent in the treatment of 
metastasizing pancreatic islet cell tumors and 
other malignancies [27] and its effect can be 
seen within seventy-two hours after 
administration depending on doses administered 
[28]. STZ has been one of the chemical agents 
used for the induction of diabetes mellitus in 
experimental animals. STZ functions as DNA-
synthesis inhibitor in bacterial and mammalian 
cells [29]. The selective pancreatic beta cell 
toxicity and diabetic condition, resulting from STZ 
induction, is related to the glucose moiety in its 
chemical structure which enables STZ to enter 
the beta cell via the low affinity glucose -2- 
transporter in the plasma membrane [30].  
 
Since diabetes mellitus is said to also cause 
decline of fertility in males [5], semen analysis is 
therefore necessary in this study as it serves as 
a pivotal and indispensable procedure in the 

assessment of male fertility status [31]. When 
conducted meticulously and in an in-depth 
manner, it possesses the ability to elucidate the 
underlying causes contributing to male infertility, 
thereby offering valuable insights and diagnostic 
clarity [32]. The comprehensive evaluation of 
semen encompasses the analysis of diverse 
parameters, including the measurement of 
ejaculate volume, quantification of sperm count, 
assessment of sperm motility and movement 
patterns, evaluation of sperm morphology and 
structural integrity, as well as the investigation of 
the composition and makeup of seminal 
secretions [32].  
  

2. MATERIALS AND METHODS 
 

2.1 Plant Collection and Extract 
Preparation 

 
Rhizomes of Curcuma longa (Tumeric) were 
bought at watt market in Calabar, Cross River 
State, Nigeria. It was identified and authenticated 
by a taxonomist in the Department of Botany, 
University of Calabar, Cross River state, Nigeria 
and a voucher number (Bot/Herb/UCC/201) was 
given. The fresh turmeric rhizomes were 
cleaned, chopped into tiny pieces and air dried 
for 7days after which they were grounded into 
powdered form. A measured amount of 950g of 
powered rhizomes were extracted using 2 liters 
of 95% ethanol for 24 hours. The extract was first 
double filtered with Chess cloth, then with filter 
paper (Whatman No.1 filter paper). The filtrate 
(extract) was concentrated under reduced 
pressure at 45oC in rotary evaporator to 10% 
volume and then dried in a vacuum water bath, 
yielding 58.9g (6.2%) of crude extract. The crude 
extract (paste) obtained was stored in a 
refrigerator until it was required. 
 

2.2 Experimental Animals 
 

Twenty-four (24) adult male Wistar rats with 
average weight of 160g were used for this 
research. The rats were kept in clean cages and 
divided into four groups designated A, B, C and 
D with six rats in each group. The rats were 
allowed to acclimatize for two weeks in animal 
house, University of Calabar and allowed access 
to commercially available chow (livestock feed) 
and water.  
 

2.3 Experimental Design 
 

Table 1 shows the experimental design of the 
research. Twenty-four animals were divided 



 
 
 
 

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randomly into four groups containing six rats 
each. The groups as shown in the table                 
includes the normal control, diabetic control, 
standard group and the test (C. longa treated) 
group. 
 

2.4 Induction of Hyperglycemia 
 
STZ was administered intra-peritoneally to the 
rats after fasting for twelve hours in order to 
induce hyperglycemia. The STZ was 
reconstituted in 0.5M sodium citrate and 
administered at a dose of 65mg/kg bw [33].   
 

2.5 Confirmation of Diabetes Mellitus 
 
Diabetes was confirmed three days after 
administration of STZ using Accu-Check 
glucometer with blood samples obtained from 
tails of the Wistar rats. The blood glucose levels 
(mg/dl) was checked before and after induction 
and every seven (7) days during administration 
of the ethanolic extract of Curcuma longa to 
ascertain hyperglycemic state. It was observed 
that the blood glucose of all the animals in the 
diabetic groups were above 170 mg/dl as 
compared to that of the normal control which had 
values less than 91 mg/dl. 
 
Administration of extract: Curcuma longa 
extract administration commenced                              
three days after induction of hyperglycemia by 
oro-gastric intubation which lasted for 28                
days. 
 
Termination of experiment: At the end of 
treatment period, the experimental animals were 
weighed and sacrificed through chloroform 
inhalation. The anterior abdominal wall was 
incised and semen was extracted from the 
caudal epididymis for analysis. The testis was 
then obtained, weighed and preserved in 10% 
formal saline for tissue processing using 
hematoxylin and eosin staining method.  
 
Determination of weight: All the animals were 
weighed before and after induction of 
hyperglycemia and every three days                       
during the course of administration of the extract.
  
Analysis of sperm parameters: The epididymis 
was dissected out and placed in a physiological 
saline in a ratio of 1:10 weight (g) by volume (ml), 
and was then macerated using surgical blade to 
release the sperm cells. The suspension was 
then filtered with 80 µm stainless mesh after 

pipetting [34]. The following sperm parameters 
were estimated as follows: 
 

i) Sperm motility (%): Two drops of sperm 
suspension were placed on a clean and 
labelled microscope slide and covered with 
a coverslip. This was mounted on light 
microscope and the number of motile cells 
divided by the total number of sperm cells 
counted and was expressed in percentage. 

ii) Sperm viability (%): This was done using 
the eosin-nigrosin staining technique. The 
sperm suspension was mixed with equal 
volume of the stain and smeared on glass 
slides. Live sperm excluded the stain and                             
appeared lightly colored, while                       
dead sperm took up the stain and 
appeared pink in color. The counts of               
live sperm were divided by the total 
number of sperm cells and expressed as 
percentage. 

iii) Sperm count (x106/ml): Improved 
neubauer hemocytometer was used for the 
sperm count. A capillary tube was used to 
pipette the sperm suspension into the 
counting chamber of the hemocytometer. 
The hemocytometer was placed on a light 
microscope and the counting was 
estimated by multiplying the number of 
cells counted by the dilution factor and by 
the hemocytometer volume. 

 
Histological study using Hematoxylin and 
Eosin stain “H&E”: The paraffin slides 
containing testicular tissue underwent a 
dewaxing process involving two rounds of 
exposure to xylene for a duration of 5 minutes 
each. Subsequently, rehydration was performed 
by sequentially immersing the slides in 
decreasing concentrations of alcohol (100%, 
95%, and 70%) and rinsing them under tap 
water. Following rehydration, the sections were 
subjected to a 15-minute staining procedure 
using hematoxylin, followed by a 5-minute rinse 
under tap water. To enhance visualization, the 
sections were then differentiated in acid alcohol 
for 1 minute and subsequently counter-stained 
with eosin for another 1 minute. After a rinse in 
tap water, the sections underwent dehydration 
and clearing using xylene. Following this, the 
sections were allowed to air dry, and a few drops 
of DPX were applied to the slide surface before 
placing a coverslip on top. The resulting tissue 
units were then taken using a digital camera 
connected to a light microscope for further 
examination [35]. 

 



 
 
 
 

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Table 1. Experimental design 
 

Groups  Number of rats Treatment 

A 6 Normal control (rat chow and distilled water) 
B 6 Diabetic control 65mg/kg bw of STZ 
C 6 65mg/kg bw of STZ + 250mg/kg.bw metformin 
D 6 65mg/kg bw of STZ + 500mg/kg.bw Curcuma longa 

 

2.6 Statistical Analysis 
 
Data obtained from the experiment was analyzed 
using one-way Analysis of Variance and Duncan 
post hoc test using a Statistical Package for 
Social Science, SPSS version 26.0 for Windows. 
The results were presented as mean ± standard 
error of mean and considered statistically 
significant at p<0.05. 
 

3. RESULTS 
 
Assessment of blood glucose: The changes in 
fasting blood glucose “FBG” were determined in 
this study over the course of the 28-day 
experimental period and they are presented in 

Fig. 1. At day zero, the blood glucose levels in all 
experimental groups were considered normal 
ranging from about 70 mg/dl to 107 mg/dl. From 
the results, it can be observed that elevated 
blood glucose concentration was seen in all 
diabetic groups following the administration of 
65mg/kg body weight of STZ. The elevated FBG 
level (Fig. 1) of the diabetic control group only 
exhibited an increase (p<0.05) of -
24.40±26.61mg/dl (gotten by subtracting the final 
FBG level from the initial FBG level after 
induction with diabetes mellitus), while that of the 
metformin and extract treated groups had 
significantly decreased FBG levels of -
227.60±24.32mg/dl and -236.60±25.14mg/dl 
respectively (p<0.05). 

 

 
 

Fig. 1. Daily blood glucose levels of the different experimental groups 
Values are expressed as mean +SEM, n = 6 

* = significantly different from normal control at p<0.05 
 



 
 
 
 

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Fig. 2. Initial and final body weights of the different experimental 
Values are expressed as mean +SEM, n = 6. 

* = significantly different from control at p<0.05 
a = significantly different from diabetic control at p<0.05 

 

 
 

Fig. 3. Sperm count in the different experimental groups 
Values are expressed in Mean + SEM. N = 6 

* = Values are significantly decreased compared to Normal Control (p<0.05) 
a = Values are significantly increased compared to Diabetic Control at p<0.05 



 
 
 
 

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Fig. 4. Sperm motility in the different experimental groups 
Values are expressed in Mean + SEM. N = 6 

* = Values are significantly decreased compared to Normal Control (p<0.05) 
a = Values are significantly increased compared to Diabetic Control (p<0.05) 

 
Assessment of body weight: The results of 
changes in the body weight of experimental 
animals after 28 days’ period were assessed. 
The initial and final weights across the 28-day 
period are presented on Fig. 2. Observed from 
these results was a significant (p<0.05) reduction 
in body weight of the experimental rats induced 
with diabetes (-37.00±5.75g) after subtracting the 
final weight from the initial weight of the rats. The 
normal control group A had a significant increase 
of +20.00±1.14g. On treatment with the test drug 
(metformin), a weight increase of +0.80±1.80g 
was observed, while that of the Curcuma longa 
extract was a decrease of -1.20±2.08g (p<0.05) 
 
Semen analysis: 
 
Sperm count: Results showed a significant 
(p<0.05) decrease of 37.70 + 7.5 million/ml and 
47.10 + 1.7 million/ml of sperm count in the 
diabetic control and metformin treated groups 
respectively compared to the normal control 
group that had 69.6 + 1.5 million/ml sperm count. 
However, 65.40 + 1.4 million/ml of sperm count 
was recorded in Group D animals (administered 
with 500mg/kg.bw of C. longa). This increase in 

sperm concentration was statistically significant 
when compared with the diabetic control 
(p<0.05) (Fig. 3).  
 
Sperm motility: Fig. 4 shows the result of sperm 
motility for the experimental animals. Animals in 
the diabetic control group and the group treated 
with metformin recorded significant decrease in 
sperm motility values when compared with the 
normal control group (p<0.05). However, Group 
D animals (placed on 500mg/kg.bw of C. longa), 
showed an increase in sperm motility which was 
statistically significant when compared with the 
diabetic control and the metformin treated             
group. 
 
Sperm viability: Experimental animals in the 
diabetic control group and the group placed on 
metformin recorded low sperm viability values 
which was significantly decreased when 
compared with the normal control group 
(p<0.05). However, Group D animals (placed on 
500mg/kg.bw of C. longa), showed an increase 
in sperm viability of which was statistically 
significant when compared with the diabetic 
control (Fig. 5).  



 
 
 
 

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Fig. 5. sperm Viability in the different experimental groups 
Values are expressed in Mean + SEM. N = 6 

* = Values are significantly decreased when compared to Normal Control at p<0.05 
a = Values are significantly increased when compared to Diabetic Control at p<0.05 

 

 
 

Plate 1. Testis (normal control) X400 
Section of testes showing normal seminiferous tubules with germinal cell layers (double-headed arrow) on the 
basal lamina to spermatocytes filled lumen (dotted circular lines). The interstitial connective (#) tissue was well 

preserved 



 
 
 
 

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Plate 2. H&E section of testis of diabetic control (X400) 
Section of testes showing seminiferous tubules with reduced germinal epithelium (double-headed arrow), 

distorted interstitial connective tissue (#) 
 

 
 

Plate 3. H&E section of testis (Standard group) X400 
Section of testes showing normal seminiferous tubules tubules with germinal cell layers (double-headed arrow) 
starting from spermatogonia (arrowhead) on the basal lamina and spermatocytes filled lumen (dotted circular 

lines). The interstitial connective (#) tissues was distorted 

 



 
 
 
 

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Plate 4. H&E section of testis (treated with 500mg/kg C.longa) X400 
Section showed different stages of seminiferous tubules and the interstices. The seminiferous with germinal cell 

layers (double-headed arrows) showed lumen partly &completely filled (dotted circular line) with mature 
spermatocytes. The interstitial connective (#) tissues was distorted. Although the alteration exists, there is a 

progressive restoration observed 
 

Histological observations: Histological study 
on sections of testes in the normal control group 
(X400) showed prominent seminiferous tubules 
with regular and consistent cellular arrangements 
on the germinal epithelium. The lumens of 
seminiferous tubules were filled with flagella of 
newly formed spermatozoa. Their basement 
membranes were intact and the intervening 
interstitium which contained blood vessels and 
clusters of Leydig cells showed regular pattern 
(Plate 1). 
 
Section of testes from the diabetic control group 
(Group B) showed significant alteration in 
histological patterns in the testes when 
compared with the normal control. Irregular 
shaped seminiferous tubules with shrunken 
lumen and decreased tubular diameter were 
observed (Plate 2).  
 
Section of testes in the diabetic animals treated 
with metformin (Group C) showed normal 
seminiferous tubules although its germinal 
epithelium was distorted with differentiating cells 

arranged in a coarse pattern. The interstitial 
connective tissue was distorted (plate 3). 

 
For the test group treated with 500mg/kg.bw of 
C. longa, seminiferous tubules were prominent 
and intact with basement membrane. The 
interstitial connective tissue was distorted, but 
there is progressive restoration observed. 
 

4. DISCUSSION 
 
Complex interplay of various pathological 
processes, prominently characterized by 
oxidative stress damage, inflammation, and 
apoptosis are involved in diabetic testicular 
damage [36].  Curcumin, a bioactive compound 
derived from Curcuma longa, has emerged as a 
potential therapeutic agent with protective 
properties against diabetic testicular injury. 
However, the precise underlying mechanisms 
through which curcumin exerts its beneficial 
effects in this context remain a subject of 
ongoing investigation. The present study 
examined the effect of C. longa crude extract on 



 
 
 
 

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the histology of the testis and semen parameters 
of streptozotocin-induced hyperglycemic male 
rats. 
 

The sustained increase in the blood glucose 
levels of diabetic animals observed at the end of 
the study was corroborated by studies by Pulido-
Moran et al. [37] and Mohammed et al. [38] 
which revealed hyperglycemic response in STZ 
treated animals. They postulated that the 
observation was due to impaired glucose 
oxidation which causes NAD+ depletion, 
ultimately culminating in the inhibition of insulin 
biosynthesis and secretion. The reduction in 
blood glucose levels in the metformin and 
extract-treated groups, aligns with the study by 
Akpaso et al. [39], who reported a significant 
reduction in fasting blood glucose levels in 
animals administered curcumin. Studies by 
Shenoy [40] and Kunnumakkara et al. [21] have 
corroborated the anti-diabetic efficacy of the 
extract, attributing it to its antioxidant and anti-
inflammatory properties. Findings reported by 
Akpaso et al. [41] also revealed the potential of 
C. longa to significantly reduce in blood glucose 
levels in experimental animals. 
 

The final bodyweight of the experimental animals 
in the extract-treated group showed a significant 
increase when compared to both the diabetic and 
control groups. The overall changes in 
bodyweight statistically indicated that both the 
metformin and extract-treated groups 
experienced significant increases in their 
respective bodyweights when compared to both 
the control and diabetic groups. These findings 
are consistent with studies conducted by Shenoy 
[42] and Szudelski [43] which reported a 
significant decrease in bodyweight in 
streptozotocin-injected rats. Furthermore, the 
observed decrease in bodyweight of the diabetic 
rats aligns with the findings of Ding et al. [44] and 
Zhong et al. [45] who observed a similar effect on 
diabetic animals induced with streptozotocin. The 
underlying mechanisms contributing to the 
reduction in bodyweight within the diabetic group 
can be attributed to the intricate interplay of 
factors such as the degradation of structural 
proteins and muscle wasting as reported by He 
et al. [46]. Conversely, the ameliorative effects of 
Curcuma longa on bodyweight is supported in 
the work of Akpaso et al. [41] where the oral 
administration of the plant extract demonstrated 
remarkable improvement in the bodyweight of 
experimental animals.  
 

Semen analysis revealed a significant reduction 
in the total count of sperm cells in the diabetic 

control group, compared to the normal control 
group. This observation underscores the 
detrimental impact of diabetes mellitus, 
regardless of whether it is type 1 or type 2 
diabetes, on male fertility. The findings of Tsao et 
al. [47] demonstrated the adverse effects of 
experimentally induced diabetes on sperm 
parameters. Also a study conducted by Alizadeh 
et al. [48] showed significantly reduced sperm 
count in diabetic animals. While the scientific 
basis underlying the pathophysiological 
mechanisms linking diabetes to semen 
parameters, particularly sperm count, remains 
diverse in the literature, prolonged hyperglycemia 
triggers the body's oxidative stress response and 
leads to endothelial injury in blood vessels, 
including those within the testis and epididymis 
[49].  Moreover, elevated blood glucose levels 
can disrupt the regulatory function of the 
hypothalamic-pituitary-gonadal axis, resulting in 
alterations in the number and morphology of 
testicular interstitial cells, degeneration of Sertoli 
cells, reduced synthesis and secretion of 
testosterone, impaired sperm development and 
maturation, and ultimately, compromised 
reproductive functions. However, the 
administration of the extract in the treated group 
led to a notable improvement in the total sperm 
count, which was comparable to that observed in 
the normal control group. These findings are in 
line with the research conducted by Chanapiwat 

et al. [50] who demonstrated that replacing a 
ketogenic diet with curcumin supplementation 
improved semen quality. Additionally, a different 
study reported an increase in total sperm count 
among infertile men participating in a randomized 
clinical trial after receiving curcumin [51]. 
Together, these studies provide further support 
for the positive effects of curcumin and its 
potential in ameliorating the adverse impacts of 
diabetes on semen parameters, including sperm 
count. 
 
The experimental findings revealed a significant 
decline in the percentage concentration of motile 
sperm cells in the diabetic group compared to the 
control group. However both the standard and 
test groups showed an increase (p<0.05) 
compared to the normal control group.  These 
observed outcome in the diabetic group is 
consistent with the outcomes of prior 
investigations conducted by Ricci et al. [52] and 
Zha [53] who reported that the addition of 
curcumin in frozen-thawed Angora goat semen 
improved both motility and acrosome integrity, 
while also enhancing the motility and functional 
integrity of sperm plasma membrane in frozen 



 
 
 
 

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bull semen. In a report by Ozbek, he stated that 
diabetes mellitus is capable of disrupting 
endocrine and metabolic pathways, leading to 
reduced sperm motility and altered sperm 
morphology [54]. The reduced sperm motility 
observed in the standard group agrees with 
studies by Raghif [55] and Naglaa et al. [56] 
which illustrated that metformin administration 
led to decrease in sperm motility. It was stated 
that vitamin B12 deficiency could be the cause of 
this observation as chronic metformin use is 
associated with lower blood levels of vitamin B12. 
Furthermore, metformin administration was said 
to impair the activity of mitochondrial complex-1 
which plays a vital role in maintaining the 
normalcy of sperm motility [56,56]. 
 

Sperm viability values which decreased 
significantly in the diabetic control and 
metformin-treated groups compared with the 
normal control group. In the extract treated group 
there was an increase in sperm viability which 
was statistically significant when compared with 
the diabetic control. This finding aligns with a 
report by Ozbek also showed that diabetes 
mellitus was capable of altering sperm 
morphology through the disruption of endocrine 
and metabolic pathways [54]. A study by Belhan 
et al [57] showed that the percentage of sperm 
viability in STZ-induced diabetic rats improved 
mildly in Curcuma longa treated group. The 
reason for the increase in sperm count, motility 
and viability as seen in the group treated with 
Curcuma longa extract may be due to its 
composite antioxidants which help reduce the 
oxidative stress caused by diabetes.                      
Oxidative stress is capable of damaging                 
sperm cells, as well as impairing their function 
[58].    
  

The histopathological manifestations in the tissue 
sections of group B (diabetic control) rats were 
characterized by irregularly shaped seminiferous 
tubules exhibiting a shrunken lumen and 
decreased tubular diameter, which were 
indicative of severe edema and pronounced 
hyperemia within the veins. These findings align 
harmoniously with the seminal research 
conducted by Ghosh et al. [59] and Barsiah et al. 
[60] who expounded upon the frequent 
occurrence of abnormal histology within the 
testes of diabetic animals, with discernible 
alterations observed in the cytoarchitecture of the 
seminiferous epithelium as well as disruptions in 
the occlusive distribution pattern.  
 

Conversely, the tissue sections derived from the 
extract-treated group displayed notable 

therapeutic effects, which include prominent 
seminiferous tubules characterized by prominent 
basement membranes. Studies by Sudjarwo et 
al. [61] and Singh et al. [62] documented the 
positive influence of Curcuma longa on the 
cytoarchitecture of the testes. This remarkable 
therapeutic outcome can be attributed to the 
phenol compounds present in the extract as 
suggested by Sadhwani [63] and Pharm-Huy et 
al. [64] that phenols possess the ability to 
counteract excessive free radicals, thereby 
safeguarding cells and tissues against their toxic 
effects. Many studies have also shown the 
tissue-protective effects of extracts of various 
medicinal plants rich in antioxidants 
[65,66,67,68,69]. 
 

5. CONCLUSION 
 
The findings of the study suggest that extract of 
Curcuma longa has the potential to improve 
fertility in diabetic male animals, as well as 
protect against the detrimental effects of 
hyperglycemia on the male reproductive system.  

 
DISCLAIMER (ARTIFICIAL INTELLIGENCE) 
 
Author(s) hereby declare that NO generative AI 
technologies such as Large Language Models 
(ChatGPT, COPILOT, etc) and text-to-image 
generators have been used during writing or 
editing of this manuscript. 
 

CONSENT  
 
It is not applicable. 
 

ETHICAL APPROVAL 
 
An approval for the experimental protocol of this 
study with registration number 220ANA2423 was 
obtained from the Faculty Animal Research 
Ethics committee (FAREC-FBMS), Faculty of 
Basic Medical Sciences, University of Calabar, 
Cross River state, Nigeria. 
 

COMPETING INTERESTS 
 
Authors have declared that no competing 
interests exist. 
 

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