# Therapeutic Effects of Lycopene Alone or in Combination with Cephalexin on Chronic Prostatitis Caused by Staphylococcus aureus in a Rat Model

Soumia Keddari, Djahira Hamed, Abla Bouhend, Mokhtaria Yasmina Boufadi, Meriem Mokhtar, Bouasria Benbouziane, Nabil Touzout, Sabrina Lekmine, Jie Zhang, Abdeltif Amrane, et al.

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► To cite this version:

Soumia Keddari, Djahira Hamed, Abla Bouhend, Mokhtaria Yasmina Boufadi, Meriem Mokhtar, et al. Therapeutic Effects of Lycopene Alone or in Combination with Cephalexin on Chronic Prostatitis Caused by Staphylococcus aureus in a Rat Model. Microbiology Research, 2025, 16 (6), pp.114. 10.3390/microbiolres16060114. [https://doi.org/10.3390/microbiolres16060114](https://doi.org/10.3390/microbiolres16060114). [hal-05155855](https://HAL.inistre.fr/hal-05155855)

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1 Article

Antibacterial effects of lycopene alone or in
combination with cephalexin on chronic prostatitis
caused by Staphylococcus aureus in a rat model

Soumia Keddari¹, Djahira Hamed¹,², Abla Bouhend¹, Mokhtaria Yasmina Boufadi¹, Meriem Moukhtar¹, Bouasria Benbouziane¹, Nabil Touzout³,⁴, Sabrina Lekmine⁵, Jie Zhang⁶, Abdeltif Amrane⁷,*, and Hichem Tahraoui³,⁷,¶

Moukhtar¹, Bouasria Benbouziane¹, Nabil Touzout³,⁴, Sabrina Lekmine⁵, Jie Zhang⁶,

Abdeltif Amrane<sup>7,*</sup>, and Hichem Tahraoui<sup>3,7,¶</sup>

¹Laboratory of Bioeconomics, Food safety and Health, Faculty of Natural Sciences and Life, Abdelhamid Ibn Badis University of Mostaganem, BP 188, Mostaganem 27000, Algeria.

²Department of Agri-Food | Faculty of Science and Technology.University of Ain Temouchent-Belhadj Bouchaib- | PO BOX 284. 46000 | Ain Temouchent, Algeria.

³Laboratory of Materials and Environment, Faculty of Technology, University of Dr Yahia Fares, Urban Center, Medea, 26000, Algeria.

⁴Department of Agronomy, Faculty of Sciences, Pole Urban Ouzera, University of Medea, Medea, 26000, Algeria

⁵Biotechnology, Water, Environment and Health Laboratory, Abbes Laghrour University, Khenchela, 40000, Algeria.

⁶School of Engineering, Merz Court, Newcastle University, Newcastle upon Tyne NE1 7RU, UK.

⁷Ecole Nationale Supérieure de Chimie de Rennes, CNRS, ISCR—UMR6226, University Rennes, 35000 Rennes, France.

*Corresponding author: hichemm.tahraouii@gmail.com ;
abdeltif.amrane@univ-rennes.fr

Abstract

Chronic bacterial prostatitis (CBP) is caused by bacterial infection,
commonly treated with fluoroquinolones. Due to rising antibiotic resistance,
alternative therapies such as phytotherapy are explored. Lycopene, a
potential antioxidant with anti-inflammatory properties, is a candidate for
such therapy. This study aims to evaluate lycopene's therapeutic effects
alone or with cephalexin against chronic prostate infections induced by
*Staphylococcus aureus* using the Wistar rat model. The CBP model was
established by introducing the *S. aureus* through the urethra into the
prostatic duct in 25 rats, confirming infection via uriculture and
spermoculture analysis. Infected rats (n=21) were grouped randomly: G1:
(control), G2 (lycopene), G3 (cephalexin), and G4 (lycopene/cephalexin),
additionally to negative control (G5) with healthy rats. Treatments were
administered intragastrically , two times per day for 2 weeks: lycopene (10
mg/kg), cephalexin (2.5 mg/kg), or both. Biological samples (blood, urine,
and prostate specimens) were collected for microbiological and histological
analysis. Results showed a significant reduction in bacterial counts in urine
and prostate (P < 0.01), especially in the group treated with both lycopene
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and cephalexin. This group also exhibited notable anti-inflammatory effect compared to single treatment and control groups. In conclusion, lycopene combined with cephalexin demonstrated a beneficial synergistic effect, indicating its potential as an effective treatment for CBP caused by *S. aureus*.

**Key words:** Chronic bacterial prostatitis, lycopene, cephalexin, synergy, *S. aureus*, rats.

# 1. Introduction

Chronic prostatitis is a persistent inflammation that affects the prostate gland, typically resulting from infectious (bacterial or non-bacterial) causes. Chronic prostatitis syndrome is the most common urological problem encountered in young men and is also a significant urological problem in men above 50 years of age. Prostate cancer can rapidly develop as a result of this poorly treated inflammation[1]. Chronic bacterial prostatitis (CBP) is the commonest cause of relapsing urinary tract infection (UTI) in men and is characterised by recurrent UTI and persistent pathogenic bacteria in the prostatic secretions[2], leading to painful symptoms, urinary and sexual problems, and a significant reduction in quality of life and psychological state[3]. Bacterial prostatitis is most commonly caused by infection with gram-negative pathogenic microorganisms, principally members of the Enterobacteriaceae family, but organisms from other families can be responsible and are more likely in certain high-risk populations. *Escherichia coli* (*E. coli*) is the most common isolate from urine cultures and is the causative agent in the majority (approximately 50% to 90%) of cases. Othercommon isolates include the species *Proteus*, *Klebsiella*, *Enterobacter*, *Serratia*, and *Pseudomonas*. Gram-positive organisms such as *Enterococcus* species[4]. One of the main bacterial problems facing public health today is *Staphylococcus aureus* (*S. Aureus*), which is most common in those suffering from urinary tract infections. While *S. aureus* has been linked to 0.5–6% of urinary tract infections biofilm forming *S. aureus* in urinary tract Infection are thought to be responsible for up to 65% of infections and 80% of chronic illnesses. Urologyis one of the key specialties where biofilm formation might pose a significant threat. Furthermore, the development of biofilms by strains responsible for acute prostatitis enhances their capacity to endure inside the prostatic secretory system, resulting in the recurring urinary tract infections that are typical of chronic bacterial prostatitis [5].

Nevertheless, long-term therapy has been recommended to eradicate the causative organism from the prostate and prevent relapses, especially if they are symptomatic. Long-term administration of antibiotics leads to potential adverse effects such as gastrointestinal, cardiovascular and skin problems and the development of bacterial resistance. Although antibiotics such as fluoroquinolones and cefalexin are used, recurrence rates are high due to the formation of biofilms, poor penetration of the drugs into the prostate and the emergence of bacterial resistance[6]. Recently, due to the lack of proven effectiveness of conventional antibiotic therapy, the goal of treatment has
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shifted from complete healing to improving quality of life and attention has been paid to herbal and other alternative treatments.

Many researchers have been interested in important phytochemicals such as lycopene, an antioxidant present throughout certain red fruits and vegetables including tomatoes and watermelon. Several functions of lycopene, including antioxidant[7], anti-inflammatory and antiproliferative, are preventive or curative for heart failure, neoplasms and therapeutic cancers[8], and protective effects of lycopene by inhibition of oxidative stress[9]. The synergistic interaction of lycopene with other drugs and micronutrients is particularly emphasized because of its binding activity with dietary components crucial in cancer prevention [1].

Lycopene typical consumption in humans (intake: 0.6 to 10.9 mg/day)[10], and the adsorption of lycopene with a dose of 10 mg, was more ideal in comparison with other high doses (30 and 50 mg) [11]. Recent research have demonstrated the correlation among prostatitis, chronic oxidative stress, and lycopene. Other investigates show that the combination of the antibiotic ciprofloxacin and lycopene improves the effectiveness for treating *E. coli* induced chronic prostate infections in Wistar rat model[2]. The purpose of our research is to investigate the synergistic interaction between lycopene and cephalexin, a β-lactam antibiotic from the cephalosporin class, on chronic prostatitis induced by the gram-positive bacterium *S. aureus* in Wistar rats.

The main objective of this experimental study is to thoroughly evaluate the therapeutic potential of lycopene, a carotenoid with well-documented antioxidant properties, administered alone or in combination with an antibiotic of the cephalosporin family, cephalexin. This evaluation is based on a rigorously established murine model of chronic bacterial prostatitis, experimentally induced by the inoculation of the pathogenic strain *S. aureus*. The study aims to determine the impact of these treatments on the histopathological, microbiological and biochemical parameters associated with chronic prostatic inflammation, while exploring underlying mechanisms of action, including those related to oxidative stress, immune response and bacterial load.

## 2. Materials and methods

### 2.1. *The chemicals and reagents used in the study included :*

Lycopene (purity ≥ 90%), purchased from Sigma-Aldrich; Medetomidine (Domitor®, Orion Pharma, Espoo, Finland); Cephalexin, obtained from LaboratoireHupPharma; and Ketamine (Imalgene® 1000, Merial, Lyon, France).

### 2.2. *Animals*

This experiment utilized 30 male adult Wistar rats, each weighing between 200 and 300 grams, 5 healthy rats were used as negative control, and 25 rats were used to induce chronic bacterial prostatitis (CBP). The animals were maintained under regular environmental conditions at 22°C under a 12 h light and 12 h dark, with food and water provided *ad libitum* [12]. The study was conducted in accordance with the National Institutes of Health (NIH) guidelines. All experiments involving animals received approval from the
137 local ethical council for animal care at the institution (University, Abdel
138 Hamid Ibn Badis, Mostaganem, Algeria) (rat/mouse 20% upkeep, RN-01-
139 20K12; Carfil Quality).

140 2.3 CBP model

141 A developed CBP rat model was carried out according to a procedure noted
142 in the literature [2]. The strain used in this experiment is *S. aureus*, a gram-
143 positive strain. This sample was acquired from a sperm culture of a patient
144 suffering from chronic prostatitis from the private clinical laboratory
145 "Bouziane", located in Mostaganem, Algeria. The isolated strain was
146 identified using the automated VITEK® system, which provided a
147 comprehensive antibiogram. The collected strain was then cultured for 18
148 hours in Brain Heart Infusion Broth (BHIB) at 37°C in an incubator. The S.
149 aureus cells were wrung out, rinsed three times and resuspended in
150 physiological solution to have a concentration of 10⁸ CFU/ml. Animals were
151 put under anesthesia with ketamine intramuscular injection (50 mg/kg), the
152 vaginal region was then disinfected with 70% alcohol, a pediatric catheter
153 (0.9 mm outer diameter, 2.5 cm length) was subsequently placed into the
154 urethra (Fig. 1). 0.2 ml of a S. aureus suspension, was injected into the
155 prostatic urethra using an insulin syringe. To avoid urine leakage due to rat
156 movement, anesthesia was maintained for 1 h, allowing the bacteria to reach
157 and invade the prostate. After 4 weeks, to confirm the establishment of CBP,
158 urine and sperm sampling was performed as follows:

159 2.4. Collection of urine

160 Urine was aseptically collected in a sterile environment near the Bunsen
161 burner and placed in a sterile petri dish. The urine sample was diluted in
162 phosphate buffered saline (PBS) and subsequently distributed over blood
163 agar to assess the presence or absence of S. aureus.

164 2.5. Sperm collection

165 Sperm collection in rats was conducted following the approach outlined in
166 [13] the same technique employed in cats, where ejaculation is
167 pharmacologically triggered via α-adeno-epididymal receptors that facilitate
168 the contraction of the vas deferens, trigone, and urinary sphincters upon
169 stimulation. Sampling was conducted in rats utilizing a pediatric urethral
170 catheter (a sterilized polyethylene tube with a diameter outside of 0.9 mm
171 and a length of 2.5 cm, truncated at its distal part) following the
172 administration of medetomidine at a dosage of 140 µg/Kg. The catheter was
173 evacuated with a syringe under air pressure, and the sperm was collected in
174 a sterile Eppendorf tube. Out of 25 samples collected in this investigation,
175 four rats failed to ejaculate. The collected sperm was applied directly onto
176 the slide and prepared for May-GrünwaldGiemsa staining according to [2].
177 Approximately 10 µL of the remaining sperm is cultured on blood agar to
178 determine the presence or absence of S. aureus and leukocytes.
179
<figure><img src="image_5.png" /><figcaption>**Figure 1.** Provocation of prostatitis; *S. aureus* instillation according to the protocol of [2].</figcaption></figure>

## 2.6. Animal distribution

Only rats with positive CBP were chosen for this experiment. Twenty one male Wistar rats were divided into four groups of 06 rats in control positive group and 05 rats in the experimental groups. In addition of 5 healthy rats as negative control.

**Group 01:** Control rats receive 1 ml of physiological water orally.

**Group 02:** Administered lycopene alone (10 mg/kg) from Sigma-Aldrich, dissolved in 2 ml of maize oil, via oral administration in two divided dosages daily for a duration of 2 weeks.

**Group 03:** Treated with the antibiotic cephalexin which was provided at a dose of 2.5 mg/kg, diluted in 2 ml of distilled water, via oral gavage in two divided doses daily for a duration of 2 weeks.

**Group 04:** Treated with lycopene/cephalexin antibiotic (10 mg/ kgbw of lycopene and 2.5 mg/ kg of cephalexin) which were dissolved in 2 ml of distilled water and administered through an oral administration in two divided doses daily for 2 weeks.

**Group 05 :** negative control group: Healthy rats received 1 ml of physiological water orally.

## 2.7. Animal sacrifice and specimen collection

After 2 weeks of treatment, animals were weighed and anesthetized by intramuscular injection of ketamine at a dose of 150 mg/kg bw., urine samples were recupered by cystocentesis for bacterial culture and blood samples were collected from the retroorbital region using a hematocrit capillary for Prostate Specific Antigen analysis (PSA), while the prostate was dissected rapidly. From each rat, a portion of prostate tissue was cut and sonicated in 10 mL of PBS for 10 minutes (under aseptic conditions) for quantitative microbiological study under a series of dilutions in tissue homogenate. The last portion was fixed in 10% buffered formalin to evaluate the histological study.

## 2.8. Prostate specific antigen
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PSA (Prostate Specific Antigen) is a test used to measure the levels of prostate specific antigen (PSA) in plasma. It is primarily employed in the assessment of prostate health, particularly for the diagnosis and monitoring of prostate cancer. Since PSA is a protease, its enzymatic activity can be measured using various laboratory techniques, such as ELISA or immunohistochemical analysis [14]. In this study, the analysis was conducted at the private clinical laboratory "Bouziane," located in Mostaganem, Algeria, using the ELISA technique.

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## 2.9. Histological study

Prostates were preserved in a 10% (v/v) formalin solution. The preserved tissues were dehydrated in four acetone baths, cleaned in toluene, then embedded in paraffin using a Leica TP1020. The specimens were sliced at 5 µm with a Leica RM 2135 rotating microtome. The sections were mounted on slides and stained with hematoxylin and eosin (H&E) [15].

## 2.10. Statistical study

Data are presented as mean ± SD, with a P value of <0.05 being statistically significant. Statistical analysis was conducted using one-way ANOVA, followed by Tukey's t-test for multiple comparisons. All analyses were conducted using Past3.exe program (version 2018).

# 3. Results

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## 3.1. CBP detection

According to the microbiological results, out of 25 rats, 21 (84%) exhibited chronic bacterial prostatitis. The presence of *S. aureus* with beta hemolysis around each colony on blood agar as well as a GRAM staining confirms the presence of this bacterium.

Sperm staining using May-GrünwaldGiemsarevaled the presence of a significant number of leukocytes in all 21 rats (Fig. 2). Four rats that did not ejaculate were excluded from the experiment.

<figure><img src="image_7.png" /><figcaption>**Figure 2.** Microscopic observation of a sperm smear from rat with chronic bacterial prostatitis (A), and a sperm from a normal rat (B) using May-GrünwaldGiems staining (at x 40 magnification). Black arrow: leukocytes cells.</figcaption></figure>

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3.2. Body weight evolution

Body and prostate weight, which enregestred at the end of the experiment, revailed a diffrente change ( Tabel 01). It was observed that lycopene and cephalexin treatment alone, 218.94 ± 25.95 g and 218.94 ± 25.95 g, respectively, induced a significant increase in body weight when compares to CBP control one (171.4 ±38.66 g). In addition, lycopene and cephalexin combination treatrement (275.34 ± 11.47 g) was found to exhibited a high significant increase in the body weight compared the the control group. Prostate weight in control and lycopene groups (0.34 ± 0.01 and 0.47 ± 0.12 g respectively) reduced significaly compared to other groups. This decrease may explained by atrophy in prostatic tissues.

Table 1. Body and prostate weight in different groups presented in mean ± standartdiviation.

<table><thead><tr><th>Groups</th><th>Body weight (g)</th><th>Prostate weight (g)</th></tr></thead><tbody><tr><td>Negative Control group</td><td>276.24 ± 11.47<sup>b</sup></td><td>0.54± 0.05</td></tr><tr><td>Positive Control group</td><td>171.4 ±38.66</td><td>0.34 ± 0.01<sup>a</sup></td></tr><tr><td>Lycopene group</td><td>218.94 ± 25.95<sup>a</sup></td><td>0.47± 0.12<sup>a</sup></td></tr><tr><td>Cephalexin group</td><td>222.5 ± 11.46<sup>a</sup></td><td>0.53± 0.05</td></tr><tr><td>Lycopene / cephalexin</td><td>275.34 ± 11.47<sup>b</sup></td><td>0.55± 0.11</td></tr></tbody></table>

a : Significant difference at P < 0.05; b : Highly significant difference at P < 0.001.

3.3. Prostate Specific Antigen analysis

No significant difference (P > 0.05) in Prostate Specific Antigen concentration in both treated and untreated animals, PSA values ranged from 2.84±0.008× 10⁻² ng/mL and 2.62±0.003× 10⁻² ng/mL (The result shown in Table 2).

Table 2. Prostate Specific Antigen parameters in in different groups presented in mean ± standard diviation.

<table><thead><tr><th>Groups</th><th>PSA (× 10<sup>-2</sup> ng/mL)</th></tr></thead><tbody><tr><td>Negative Control group</td><td>2.83 ± 0.002*</td></tr><tr><td>Positive Control group</td><td>2.64 ±0.008*</td></tr><tr><td>Lycopene group</td><td>2.66 ± 0.003*</td></tr><tr><td>Cephalexin group</td><td>2.62± 0.004*</td></tr><tr><td>Lycopene / cephalexin</td><td>2.84 ± 0.001*</td></tr></tbody></table>

3.4. Prestate and urine cultures

The quantitative microbiological result of both prostate tissues and urine cultures of rats showed that there was a highly significant decrease in colonies number (CFU) in the cephalexin and lycopene/cephalexin groups
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compared to the control one (P < 0.05) (Table 03). A decrease no significant (P > 0.05) in CFU number in lycopene treated group recorded also compared to positive control.

<table><caption>**Table 3.** Quantitative microbiological results of urine and prostate tissue culture presented as mean ± Standard deviation.</caption><thead><tr><th>Groups</th><th>log10 CFU/gof<br>prostate tissue</th><th>log10 CFU/ 100 µl<br>of urine</th></tr></thead><tbody><tr><td>Negative control group</td><td>00.00</td><td>00.00</td></tr><tr><td>Positive Control group</td><td>6.43 ± 0.40</td><td>5.21 ± 0.16</td></tr><tr><td>Lycopene group</td><td>5.48 ± 0.36</td><td>4.66 ± 0.75</td></tr><tr><td>Cephalexin group</td><td>3.48 ± 0.37a</td><td>2.92 ± 0.89a</td></tr><tr><td>Lycopene / cephalixin</td><td>1.19 ± 0.39a</td><td>1.08 ± 0.05a</td></tr></tbody></table>

CFU, colony-forming units.a: P < 0, 05 compared with control group

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### 3.5. Prostat histological results

Agher two weeks of treatment, a several structural changes was observed in histological sections in CBP tissues compared in treated groups (Figure 03). For intrest, an intense infiltration of monocytes and macrophages, interstitial fibrosis, glandular edema was observed in the acini and interstitial tissue in CBP group (Figure 03.A). Inintrest, microscope observation in lycopene-treated group revealed a decrease in inflammatory cell infiltration in the glandular and peri-glandular tissues, mild and focal cell infiltration (Figure 03. B). Moreover, group treated with cephalixin shown a decrease of the cellular infiltration but with the presence of edema (Figure 03. C). In addition, in the group treated with lycopene/cephalexin, the acini recovered their size and their lumen contained some lymphocytes, a normal aspect of fibrous tissue and no edema (Figure 03. D).
<figure><img src="image_8.png" /><figcaption>**Figure 3.** Histological sections of prostate tissue (H &E) **A:** Negative control group of a rat with no chronic bacterial prostatitis with normal structure. **B:** control group of a rat with chronic bacterial prostatitis (CBP model) had a large number of inflammatory infiltration (II) into the intercellular substance. In addition, interstitial edema (ED), interstitial fibrosis (IF) and blood vessel congestion (BVC) were observed. **C:** Lycopene-treated group, a decrease in infiltrated inflammatory cells (II) was observed, with mild and focal cell infiltration. **D:** group treated with cephalexin, a significant decrease of cellular infiltration was observed but with the presence of edema (ED). **E:** the group treated with lycopene/cephalexin, slight lymphocytic infiltration and a normal aspect of fibrous tissue, the acini recovered their size and absence of edema.</figcaption></figure>

**4. Discussion**

Chronic prostatitis of infectious origin constitutes the highest prevalent urological disease that affects men under 50 years of age [16]. Several germs can be encountered, gram-negative bacilli are the most common germs, It has been identified that *S. aureus*, a gram-positive microorganisms, also play a role in chronic bacterial prostatitis etiology [17].

In the present work, a reproducible rats model of chronic bacterial prostatitis (CBP) was developed by introducing *S. aureus* bacteria that traveled the

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urethra and penetrated the prostatic tract inducing prostatic infection. After four weeks of inoculation 21 rats (84%) out of 25 rats developed chronic bacterial prostatitis, urine and sperm samples were taken to confirm the infection. Semen collection was performed to avoid complications of prostate biopsy surgery, especially to avoid rat death, and to improve on the protocol of [2]. Where there were 6 rat deaths due to complications following surgery. Since 80% of the sperm fluid ill elaborate from prostate, a high leukocyte count established by May-GrünwaldGiemsa staining as well as S. aureus presence in the sperm culture is sufficient to confirm prostate infection. During the two weeks of treatment, the significant decrease in body weight in untreated group compared to the treated groups caused by the loss of appetite in the rats with prostatitis. Prostate antigen (PSA) is a serine protease, elaborated by the prostatic ductal epithelium. Generally, the concentration of PSA be elevated in patients with adenocarcinoma, and even in many of the non-malignant conditions of the prostate, especially benign prostatic hyperplasia (BPH) [18]. Also, it was evaluated in bacterial prostatitis according to the studies of [16], which was noted that the concentration of PSA can increase only in the first week after infection in acute prostatitis, and then decrease and remain stable in the following two weeks (chronic prostatitis) [19]. This observation in PSA analysis is in agreement with our result. On the other hand, the quantitative microbiological study of urine and prostate tissue indicated treatement with cephalexin only or in combination with lycopene significantly decrease in colony count (CFU). It was noted that cephalexin, an antibiotic used in chronic prostatitis treatment due to S. aureus [20,21], belonging to the family of β-lactam that inhibits cell wall production and peptidoglycan synthesis. This disorder of cell structure lead to membrane permeability, so lycopene can easily cross the membrane, which reinforces their antimicrobial effect. This suggest that there is a possibility of a synergy between lycopene and the tested antibiotic.

Lycopene is well known as an antioxidant that contributes to the protection of white blood cells, immunostimulants and involving anti-inflammatory effects [22]

Chronic prostatitis, particularly when caused by S. aureus, can lead to significant histological changes in prostate tissue, including marked inflammatory cell infiltration, interstitial fibrosis, vascular congestion, and massive interstitial edema. These changes are indicative of the body's response to persistent bacterial infection and inflammation. The treatment of such conditions remains challenging due to increasing antibiotic resistance and the complex nature of the disease. This infection is characterized by significant inflammatory cell infiltration, which can lead to fibrosis in the prostate tissue. This fibrosis is associated with lower urinary tract symptoms (LUTS) and impaired urethral function, as well as the persistent of inflammation and infection inducing vascular congestion and interstitial edema[23].

This opportunistic Gram-positive pathogen (S. aureus) is able to colonize prostatic tissues by activating Toll receptors, in particular TLR2 and TLR4, expressed on immune and epithelial cells. This activation triggers the NF-κB signaling pathway, which plays a central role in initiating and maintaining inflammation by stimulating the expression of pro-inflammatory cytokines
such as TNF-α, IL-1β and IL-6. In addition, *S. aureus* is able to form biofilms on prostatic tissues, which promotes the persistence of the infection by making it more resistant to antibiotic treatments and the immune response of the host, thus contributing to the chronicity of the disease[24,25].

Lycopene is also suggested in many studies to reduce the production and expression of inflammatory factors [23, 26]. Further explained that the possible mechanism of lycopene in reducing inflammatory factors could be that lycopene was a lipophilic compound closely associated with the cell membrane, where carotenoids regulate the activities of redox sensitive transcription factors and kinases. It works by neutralizing reactive oxygen species (ROS), there by reducing tissue oxidative stress, and positively regulating the activity of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx). Lycopene also inhibits the activation of NF-κB, which reduces the production of pro-inflammatory mediators (IL-6, TNF-α, COX-2) and moderates leukocyte infiltration in tissues[27]. Other studies show that lycopene exerted an anti-inflammatory effect by down regulating the levels of inflammatory factors. And also exerts an antioxidant effect [28]. Our results and their interpretations were supported by these studies. The association of lycopene with an antibiotic treatment such as cefalexin is based on a pharmacological synergy. Cefalexin is a β-lactamine antibiotic that works by inhibiting bacterial wall synthesis, causing the lysis of bacterial cells. However, in the context of chronic prostatitis, its effectiveness may be reduced by the formation of bacterial biofilms, intense tissue inflammation and impaired permeability of prostate tissues[29]. This combined action allows for better bacterial eradication and an overall improvement of the prostate condition. Several studies have examined the efficacy of lycopene in chronic prostatitis and compared it to other natural anti-inflammatory drugs. In an animal model of chronic prostatitis/pelvic pain syndrome (CP/CPPS) [30]. [31;32] showed that lycopene significantly reduced pro-inflammatory cytokines such as TNF-α, IL-1β, IL-2 and IL-6. Another comparative study analysed the effects of lycopene alone and in combination with other natural compounds such as quercetin and curcumin, a pharmacological synergy, where the combination resulted in a more pronounced reduction of inflammatory mediators and oxidative stress than each compound administered alone. [33] showed in rats that the addition of lycopene to ciprofloxacin treatment increased the antibacterial and anti-inflammatory efficacy of the treatment, compared with the antibiotic alone. This synergistic combination not only improves treatment outcomes but may also allow for lower antibiotic doses, thus reducing the risk of side effects and antibiotic resistance

## 5. Conclusion

Based on the findings of this study, it became clear that *S. aureus* installation in prostate tissues alters body weight, endocrine homeostasis, sperm parameters and prostate histology which causes a deterioration of male reproductive function. Nonetheless, the composition of lycopene and cefalexin as a treatment of these alterations can detoxify the prostate
damage. This study highlights the importance of lycopene, as potential therapeutic natural agent to reinforce cephalexin treatment in chronic infection and improve male fuction. Further studies are necessary to identify the plausible involved mechanisms of action and the synergistic effect between this molecules. However, these preliminary results justify further research on a larger animal model, including in-depth analyses to assess the therapeutic potential of this approach.

### ACKNOWLEDGEMENT

This work is supported by Ministry of Higher Education and Scientific Research, University Abdelhamid Ibn Badis, Algeria. As a part of the Research Project (Projets de Recherche-Formation Universitaire PRFU). Code: D00L01UN270120200002 « The contribution of the intestinal microbiota to the physiological balance of the body ».

The authors would like to thank Dr Moulay Abdallah SIMERABET pharmaceutical industry laboratory manager PHARMAGHEB of Tiaret and Dr. Bentiaba Karima, for the Experiments assistance and for many helpful discussions.

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