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. 2025 Aug 22;11(5):e70578. doi: 10.1002/vms3.70578

Therapeutic Effect of Barberry (Berberis vulgaris) Root Extract on Experimental Cutaneous Leishmaniosis in a BALB/c Mouse Model

Sadegh Shirian 1,2,✉, Morteza Norouzi Cholcheh 3, Saeed Habibian Dehkordi 4, Behnam Bakhtiari Moghadam 5
PMCID: PMC12372602  PMID: 40844693

ABSTRACT

The bioactive compounds derived from plants offer promising avenues for discovering new treatments for various types of leishmaniasis. Approximately 22 alkaloids have been identified in different parts of the barberry plant, particularly in the roots. This study aimed to investigate the therapeutic effects of barberry (Berberis vulgaris) on the treatment of experimental cutaneous leishmaniasis (CL) caused by Leishmania major in a BALB/c mouse by using pathological, immunohistochemical, and clinical methods. Thirty BALB/c mice, aged 6–8 weeks, were purchased from the Razi Animal Laboratory at Karaj Institute and allowed for adaptation by 1 week. The mice were divided into three groups including control, treatment, and vehicle groups. Animals of both treatment and vehicle groups received a subcutaneous injection of 0.1 mL culture medium containing 2 × 106 promastigotes at the base of their tails. The vehicle and treatment groups received Vaseline alone and barberry root extract mixed (as ointment) in Vaseline, respectively. The intact mice were used as the control group without any intervention. After 32–35 days, the wounds were formed at the injection sites. The extract ointment was applied to treat the wounds three times daily for 28 days. The wound diameters were measured on days 7, 14, 21, and 28 post‐treatments, and the skin tissue samples were investigated using haematoxylin and eosin staining. Clinical evaluations revealed that the wound size was significantly decreased in the treatment group compared to the controls (p < 0.05). The treatment group also exhibited lower Leishmania parasite loads than other groups. Unlike the expression of TGF‐β, the mean level of IL‐1 and IL‐6 was significantly increased (p < 0.001) in the treatment group compared to other groups. These findings suggest that Berberis vulgaris can effectively improve skin lesions caused by L. major by modulating inflammatory responses in the BLAB/c mouse model.

Keywords: barberry root, cytokine, cutaneous leishmaniasis, extract

Barberry (Berberis vulgaris) root extract shows potential in treating cutaneous leishmaniasis caused by Leishmania major. In a BALB/c mouse model, treatment significantly reduced wound size and parasite load while enhancing inflammatory markers IL‐1 and IL‐6. These results highlight barberry's therapeutic promise against leishmaniasis.

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1. Introduction

According to the World Health Organization (WHO), leishmaniasis poses a significant health challenge in various regions, particularly in the Eastern Mediterranean (Fahrion et al. 2018). There are three major forms of this disease: visceral leishmaniasis (VL), cutaneous leishmaniasis (CL), and mucosal leishmaniasis (ML). Among these, cutaneous leishmaniasis has reported the highest infection rates (Askari et al. 2018; Shirian et al. 2012). In 2018, the WHO documented 100,000 new cases of leishmaniasis across these regions, including 18,175 cases of CL from Iran (Salari et al. 2020). CL is primarily caused by Leishmania major and Leishmania tropica (Piroozi et al. 2019).

In various regions of Iran, Phlebotomus papatasi has been confirmed as a vector for CL (Hanafi‐Bojd et al., 2015). Additionally, several rodent species, including Rhombomys opimus, Meriones libycus, Tatera indica, Meriones hurrianae, and Nesokia indica, have been repeatedly reported as reservoirs for L. major in different endemic foci across Iran (Abedi‐Astaneh et al. 2016; Gholamrezaei et al. 2016; Shokri et al. 2017). Furthermore, Phlebotomus sergenti also serves as a vector for CL, while humans are recognized as its reservoir (Abedi‐Astaneh et al. 2016; Norouzinezhad et al. 2016). Notably, recent reports have indicated skin lesions resembling those caused by L. infantum in various parts of Iran (Badirzadeh et al. 2013). It is estimated that ∼20,000 new cases of CL are reported annually in Iran; however, due to under‐reporting, the actual number may be significantly higher. Epidemiologically, CL is endemic in both rural and urban areas of Iran, with outbreaks occurring in both settings. The rural form of CL is present in 15 provinces, while the urban form is widespread across nearly all urban areas of the country (Sabzevari et al. 2021). This disease inflicts considerable morbidity and mortality not only in Iran but also globally (Norouzinezhad et al. 2016; Piroozi et al. 2019).

Current drug treatments for leishmaniasis include pentavalent antimonials, amphotericin B, paromomycin, miltefosine, and liposomal amphotericin B. However, the use of these medications is often limited due to factors such as low efficacy, severe side effects, high toxicity, the potential for parasite resistance, lengthy treatment regimens, and high costs. Natural products may offer a rich source of chemical diversity for discovering new therapeutic agents. Ideally, new drugs should be less toxic or non‐toxic, safe, more effective, affordable especially for low‐income populations, and readily available (Oryan 2015). Bioactive compounds derived from plants represent a promising avenue for developing new anti‐leishmanial drugs. The anti‐leishmanial properties of certain plants have been linked to compounds such as alkaloids, chalcones, triterpenoids, naphthoquinones, quinones, terpenes, steroids, lignans, saponins, and flavonoids (Lage et al. 2013; Sifaoui et al. 2014).

Barberry, scientifically known as Berberis vulgaris, is a thorny plant belonging to the Berberidaceae family (Akbar 2020). Research has identified ∼22 alkaloid compounds, including berberine, in the roots, leaves, and fruits of barberry (Arayne et al. 2007). Notably, the root bark contains a higher concentration of effective alkaloids compared to the aerial parts of the plant (Neag et al. 2018). Analyzing the chemical composition of barberry extract reveals that alkaloids with an isoquinoline structure such as protoberberine, berberamine, tetrandrine, kendocorine, and palmatine are significant components (Kalmarzi et al. 2019). Consequently, this study aims to evaluate the therapeutic effects of barberry root extract on experimental CL in a BALB/c mouse model.

2. Materials and Methods

2.1. Ethical Statement

The Ethical Committee of Shahre‐Kord University approval this study (Approval number: IR.SKU.REC.1404.014). All the experiments were done in accordance with the rights of the animals, and the animals were not harmed in the clinical trial.

2.2. Plant Collection and Extraction

The barberry plant was sourced from Qaenat farms in eastern Iran, specifically in the northern region of the South Khorasan province. The plant material was dried and crushed in a controlled environment, away from direct light and heat. Compounds were extracted using a solvent mixture of 80% methanol and water at a weight‐to‐volume ratio of 1:10 (100 g of powdered plant material in 1000 mL of solvent). Extraction was performed using a household microwave oven for 15 min at 200 watts. After irradiation, the extract was cooled, and filtration was carried out using Whatman No. 1 filter paper. To remove the solvent, a rotary evaporator was employed under vacuum conditions at temperatures ranging from 40 to 56°C, concentrating the extracts. The concentrated extracts were then dried using a freeze dryer and stored at −20°C until further use.

2.3. Preparation of Barberry Root Ointment

The ointment base was prepared using the melting method. Semi‐solid and solid components, including yellow petroleum jelly, yellow wax, and stearic alcohol, were melted together. Olive oil was incorporated into the mixture, which was heated to 70°C to ensure the complete dissolution of cholesterol. Stirring continued until the temperature decreased to 40°C. At this point, the concentrated semi‐solid extracts were gradually blended with a portion of the base in a geometric fashion before being combined with the remaining base. This mixing process continued until a uniform product was achieved. The final formulation was then packaged into suitable 15‐g tubes.

2.4. Parasite Culture

The standard strain of the L. major parasite (MRHO/IR/75/ER) was obtained from the Faculty of Medicine at Isfahan University of Medical Sciences. To propagate promastigotes in sufficient quantities, Novy‐MacNeal‐Nicolle (NNN) medium was supplemented with RPMI‐1640 medium (Aldrich Sigma), enriched with 10% fetal calf serum (Bovogen, Australia), 292 µg/mL glutamine, and 100 µg/mL penicillin‐streptomycin. The cultures were maintained at a temperature of 25°C and examined every 2 days. Once the promastigote forms reached the stationary phase, the contents of the culture tubes were centrifuged, and the sedimented parasites were washed multiple times with sterile PBS buffer. The final concentration was adjusted to 106 × 2 promastigotes per 100 µL of solution, counted using a Neubauer slide.

2.5. Clinical Intervention

Thirty BALB/c mice aged 6–8 weeks were obtained from the Karaj Institute. After a 1‐week acclimatization period in the Faculty of Veterinary Medicine's housing facility, 0.1 mL of the culture medium containing 2 × 106 promastigotes was drawn into an insulin syringe and injected subcutaneously at the base of the tail of all mice except for the control group. The mice were divided into three groups: one treated with barberry root extract, a vehicle group treated with Vaseline (Placebo), and a control group that received no treatment. All groups were housed under controlled conditions. After 32–35 days, a wound formed at the injection site. At day 35, the skin samples from the wound site were taken and stained with haematoxylin and eosin to confirm the presence of parasites under a microscope. For treatment, after restraining the mice, the ointment containing the extract was applied to the wound, and the animals were returned to their cages. Treatment was administered three times daily (morning, noon, and night) for 28 days across all groups. The first control group received Vaseline ointment at the same frequency.

2.6. Determining the Severity of Parasitic Infection

To assess the number of parasites at the lesion site, 1‐cm pieces from the lesion area were collected from three mice in each group and fixed in 10% formalin. After staining with haematoxylin and eosin, parasites were counted in 10 microscopic fields (at ×400 magnification), and the percentage reduction in each group was calculated.

2.7. Histopathological Study

Three small tissue samples from each group were collected from the lesions of euthanized mice and immediately fixed in 10% buffered formalin. The samples were then dehydrated with graded ethanol, cleared with xylene (two times each of 5 min), and impregnated with paraffin wax. The 5‐µm thick sections were routinely stained using haematoxylin‐eosin. The stained sections were blinded and studied by a light microscope (Olympus, Tokyo, Japan).

2.8. IHC Method

Sections with 3 µm thickness were used for the IHC analysis. The slides were deparaffinized first in xylol for 10 min, rehydrated in graded ethanol. After washing the slides with PBS for 5 min, they were treated with 3% hydrogen peroxide solution for 5 min at room temperature (RT) to quench the endogenous peroxides. The slides were then boiled in a microwave (power 100 for 10 min) using a 10‐mmol/L concentration of citrate buffer (pH 6.0) for the antigen retrieval step. The slides were allowed to cool and washed in PBS for 5 min. The primary antibody XLVI‐5B8‐B3(T1) (Hamburg, Germany) specific for Leishmania major was applied for 1 h (diluted 1:200). The Envision+ (DakoCytomation, Glostrup, Denmark) was used as a detection system and developed with diaminobenzidine (DakoCytomation). 3,3′‐Diaminobenzidine–hydrogen peroxide was applied as the chromogen, and haematoxylin was used as the counterstain. Ten images from each sample were taken, and the mean percentage of each protein expression was calculated by using the ImageJ software.

2.9. Serological Tests

The levels of IL‐1, IL‐6, and TGFβ on the wound surface on days 3 and 7 post treatment were measured using ELISA. The taken samples were homogenized, and the levels of aforementioned cytokines were quantified using the ELISA method. A Quantikine ELISA kit (USA) was used to measure interleukins 1 and 6, and Human TGF‐β ELISA Kit (China) was used to measure TGF‐β.

2.10. Statistical Analysis

The SPSS software was used for data analysis. The data obtained from macroscopic examinations, wound areas, and IHC staining (the mean percentage expression of each cytokine) in all groups at various time points were analyzed using one‐way analysis of variance (ANOVA) and followed by post hoc Tukey test. A p value less than 0.05 was considered statistically significant.

3. Results

3.1. Gross Examination

On the seventh day following wound formation, no significant difference was observed among the groups (p > 0.05) in terms of wound size. However, by day 14, a significant difference emerged between the placebo group and the control group (p = 0.007) in terms of wound size. Additionally, a notable difference was found between the treatment or barberry group and the placebo group (p = 0.004), with the wound diameter being smaller in the barberry group. A significant difference was also noted between the barberry group and the control group (p < 0.001) in terms of wound size.

By day 21, a significant difference was observed between the placebo group and the control group (p = 0.046), with the wound diameter smaller in the placebo group. Furthermore, the wound size in the barberry group was significantly decreased compared to the placebo and control groups (p < 0.001).

On day 28, a significant difference was recorded between the placebo group and the control group (p = 0.039). There were also significant differences between the barberry group and both placebo and control groups (p < 0.001). The wound diameter in the barberry group was lower than that of the placebo group, which in turn was smaller than that of the control group (Table 1).

TABLE 1.

Diameter of wounds based on mm2 in different groups.

Study days
Groups 7 14 21 28
Control 0.8660a ± 1.35 0.8145a ± 1.7433 a1/0 ± 1.9 2a/0 ± 2/2
Placebo 0.7234a ± 1.3767 0.0611b ± 1.4533 b06557/0 ± 1.69 0.05992b± 1.84
Barberry 0.08083a ± 1.2133 0.0755c ± 1.13 c07638/0 ± 1.0167 0.10786c ± 0.9267
p value >0.05 <0.05 <0.05 <0.05
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Note: Dissimilar letters indicate significant differences in the groups at p ≤ 0.05 (mean ± standard deviation).

3.2. Results of Histopathological Studies on Wound Healing

Immunohistochemical (IHC) staining results indicated the mean percentage of expression of IL‐1 protein was significantly increased in the barberry group compared to the placebo group (p = 0.006) and the control group (p = 0.001). The mean percentage expression of IL‐6 protein was significantly increased in the barberry group compared to the placebo group (p = 0.006) and the control group (p = 0.007). The mean percentage expression of TGF‐β protein was significantly increased in the placebo group compared to the control (p = 0.006). Additionally, higher mean percentage expression levels of TGF‐β protein were detected in the barberry group than in the control group (p = 0.007). The results of the mean percentage expression of all proteins are shown in Table 2. IHC staining images of wounds from the different treatment groups are presented in Figures 1, 2, 3, 4.

TABLE 2.

The mean percentage expression of inflammatory proteins in different groups.

Study factors
Groups IL‐1 IL‐6 TGF‐β
Control 1.29099a ± 1.5 1.41421a± 2 5.35413a ± 2.5
Placebo 2.08167a ± 5.5 1.70783b ± 1.75 1.82574b± 3
Barberry 6.55744b ± 17.5 5.12348c ± 11.25 1.73205c ± 13
p value <0.05 <0.05 <0.05
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Note: Dissimilar letters indicate significant differences in the groups at p ≤ 0.05 (mean ± standard deviation).

FIGURE 1.

FIGURE 1

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The microscopic findings obtained from skin lesions in various groups. (a) In the barberry group, mild infiltration of macrophages containing Leishman bodies is seen. The blue arrow indicates macrophage, and the yellow arrow indicates macrophage containing Leishman body. H&E staining (magnification ×400). (b) In the control group, severe infiltration of macrophages containing Leishman bodies is seen. The blue arrow indicates a macrophage, the yellow arrow indicates a macrophage containing a Leishman body, and the green arrow indicates a giant cell containing a Leishman object.

FIGURE 2.

FIGURE 2

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The mean percentage of interleukin 1 expression in different groups. (a) The mean percentage of interleukin 1 expression in barberry group. (b) The mean percentage of interleukin 1 expression in the placebo group. (c) The mean percentage of interleukin 1 expression in the control group. IHC staining (magnification ×400).

FIGURE 3.

FIGURE 3

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The mean percentage of interleukin 6 expression in different groups. (a) The mean percentage of interleukin 6 expression in barberry group. (b) The mean percentage of interleukin 6 expression in the placebo group. (c) The mean percentage of interleukin 6 expression in the control group. IHC staining (magnification ×400).

FIGURE 4.

FIGURE 4

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TGF‐β expression in different groups. (a) The mean percentage of TGF‐β expression in barberry group. (b) The mean percentage of TGF‐β expression in the placebo group. IHC staining (magnification ×400).

3.3. Results of Serological Test

A significant increase in the levels of IL‐1 and IL‐6 was detected in the the barberry extract group compared to the control and Vaseline groups on days 3 and 7 post‐treatment (p < 0.01). The results of this study showed that over time, the IL‐1 and IL‐6 levels were decreased in all groups studied. Also, on days 3 and 7 post‐treatment, the TGF‐β levels were significantly decreased in the treatment groups compared to the control group (p < 0.01). The results of this study showed that the level of TGF‐β was increased over time during wound healing (Figure 5)

FIGURE 5.

FIGURE 5

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Comparison of IL‐1β, IL‐6, and TGF‐β levels (pg/mL) in different treatment groups on days 3 and 7 post‐treatment.

4. Discussion

In this study, the mean percentage expression levels of IL‐1, IL‐6, and TGF‐β proteins in the wound created by L. major in the BALB/c model of CL before and after treatment by barberry extract were evaluated. To our knowledge, this is the first investigation into the therapeutic effects of the barberry plant on tissue cytokine levels in CL. Our results indicate that the mean percentage expression level of IL‐1 protein in the barberry‐treated group was significantly higher than those of the other groups (p < 0.05). The role of IL‐1 in promoting the development of Th1 cells in BLAB/c mice has been previously reported. For instance, one study found that tissue IL‐1 levels at the wound site significantly increase following treatment with Protargol compared to other treatments. This suggests that one potential mechanism for barberry's wound‐healing effects may involve the elevation of IL‐1 at the wound site, which could subsequently enhance Th1 cell development and facilitate the clearance of parasites. The importance of Th1 cell development in resisting leishmaniasis is well‐established, primarily through the production of pro‐inflammatory cytokines such as interleukins 1, 2, and 12; interferon‐gamma; and tumour necrosis factor. These cytokines activate macrophages and promote parasite elimination. Additionally, IL‐1 plays a critical role in regulating Th17 cell‐mediated immune differentiation. Together with IL‐6 and IL‐23, IL‐1 influences the differentiation of Th17 cells and the cytokines they produce. Mortazavi et al. (2019) highlighted that the absence of Th17 cells in patients with chronic CL can prolong the disease duration. Furthermore, Maspi et al. (2016) demonstrated that IL‐1α is effective in treating skin wounds caused by L. major. Shokri et al. (2017) identified intermediate monocytes as a primary source of IL‐1β. Overall, studies on CL in humans and animal models have shown that IL‐1 and TNF‐alpha are crucial inflammatory mediators in affected patients. These findings underscore the potential therapeutic role of barberry extract in modulating the mean percentage expression of cytokine proteins in the lesion site and enhancing immune responses against CL.

In our study, we have observed a significantly higher mean percentage expression of IL‐6 protein in the mice treated with barberry extract. IL‐6 is known to induce the production of anti‐inflammatory proteins, such as the IL‐1 receptor agonist and soluble TNF receptors. It plays a dual role in host defence against Leishmania, both enhancing and inhibiting immune responses, and has been shown to suppress INF‐γ gene expression. This cytokine also stimulates the secretion of IL‐27, which subsequently leads to increased IL‐10 production. IL‐6 may serve as a potential therapeutic target against Leishmania. In canine models, an increase in anti‐Leishmania antibody titres is typically associated with elevated levels of IL‐6. Furthermore, this cytokine appears to exert an inhibitory effect on TNF‐α in humans. Bahrami et al. (2018) found that the mRNA levels of the pro‐inflammatory cytokine TNF‐α in biopsies from CL lesions correlated with wound size. While high levels of TNF‐α in the bloodstream are recognized as biomarkers for the severity of Leishmania infection, uncertainties remain regarding its precise function. Additionally, the IL‐6 mean percentage expression is upregulated during challenges with the CL, which has been linked to lesion diameter in affected patients.

Our findings also revealed that the mean percentage expression of TGF‐β was significantly lower in the barberry‐treated group compared to the other two groups. In immunocompromised patients exhibiting immunosuppressive responses and progressing leishmaniasis, increased levels of TGF‐β have been observed. The role of TGF‐β in susceptibility to CL has been established in mouse models infected with L. amazonensis and L. chagasi, where it has been shown to facilitate an increase in parasite numbers. The decrease in TGF‐β production is linked to an increase in interferon levels. Recent findings indicate that disruption of the TGF‐β pathway leads to increased iNOS levels, exacerbating leishmaniasis (Di‐Blasi et al. 2019). Dayakar et al. (2019) showed that TGF‐β inhibits the activity of TNF‐α and INF‐γ while also regulating Th1 and Th2 responses. Unlike IL‐10, TGF‐β has a marginal effect on parasite load and host resistance associated with INF‐γ in L. donovani infections. Locally activated TGF‐β promotes parasite growth by suppressing the immune response. In animal models, TGF‐β secreted by Leishmania‐infected lymphocytes shifts the arginine pool from iNOS to arginase, resulting in polyamine production that supports parasite growth. The secretion of TGF‐β can interfere with treatment (Dayakar et al. 2019).

In our study, a 10% extract of Berberis vulgaris (barberry) was used as a topical ointment for treating wounds caused by L. major. The results indicated significant differences in wound diameter on the 14th, 21st, and 28th day between the placebo group (treated with Vaseline) and the barberry extract group, with the latter showing smaller wound diameters (p < 0.05). These findings highlight the positive effect of barberry extract on wound healing. Berberine, a four‐membered alkaloid found in nine plant families, has demonstrated proven therapeutic effects against both cutaneous and visceral forms of leishmaniasis in clinical tests. Vennerstrom et al. (1990) confirmed its potential anti‐leishmanial properties but did not elaborate on the chemical or biological mechanisms responsible for these effects (Vennerstrom et al. 1990). In a review, Parvizi et al. (2020) noted that traditional Iranian medicine treats cutaneous leishmaniasis similarly to its wet form. However, the same article described various traditional methods for addressing this general lesion without distinguishing between treatments for wet and dry forms. Traditional Iranian medicine has recommended ointments derived from boiling barberry roots for this condition (Parvizi et al. 2020).

Shahabadi et al. (2014) reported a 90% improvement using barberry root extracts in treating CL. Derived from Streptomyces rimosus subsp. containing ammonium salts (such as DMSO) was effective in both healing wounds and eliminating parasites from the host. Conversely, berberine extract also achieved complete parasite removal from the wound area; however, PR's therapeutic efficacy was found to be 20 times greater than that of berberine.

In the study by Mahmoudvand et al. (2014), both methanolic and aqueous extracts of Berberis vulgaris, along with its primary active component, berberine, demonstrated the ability to inhibit the growth of L. tropica and L. infantum promastigotes in vitro (Mahmoudvand et al. 2014). Berberine and other compounds in barberry extract have been reported to be toxic to amastigotes within macrophages, effectively reducing their population. These substances decreased the survival rate of amastigotes inside macrophages and mitigated the resulting contamination. Notably, the promastigote form is more sensitive to barberry extract compared to the amastigote form (Mahmoudvand et al. 2014). Fata et al. (1970) reported a reduction in the diameter of skin lesions caused by L. major with increasing concentrations of ethanolic extracts from the roots, stems, and leaves of the barberry plant. In this study, the alcoholic extract of barberry root proved to be more effective than those derived from its stem and leaves (Fata et al. 1970).

5. Conclusion

The findings of this study indicate that barberry can improve the skin lesions of CL induced by L. major in the BLAB/c mouse model by increasing tissue levels of interleukins 1 and 6 proteins as well as the decreasing tissue TGF‐β proteins levels. Although further research is necessary to explore therapeutic strategies based on medicinal plants for CL, it is recommended that future studies assess the effects of this plant on Th1 cells and other T‐cell populations.

Author Contributions

Norouzi Cholcheh Morteza, Sadegh Shirian: methodology. Norouzi Cholcheh Morteza, Saeed Habibin Dehkordi, Sadegh Shirian, Bakhtiari Moghadam Behnam: software. Norouzi Cholcheh Morteza, Sadegh Shirian, Bakhtiari Moghadam Behnam: data curation. Norouzi Cholcheh Morteza, Saeed Habibin Dehkordi: investigation. Norouzi Cholcheh Morteza, Saeed Habibin Dehkordi: validation. Norouzi Cholcheh Morteza, Saeed Habibin Dehkordi, Sadegh Shirian, Bakhtiari Moghadam Behnam: formal analysis. Saeed Habibin Dehkordi, Sadegh Shirian: supervision. Norouzi Cholcheh Morteza: funding acquisition. Norouzi Cholcheh Morteza, Sadegh Shirian: visualization. Sadegh Shirian: project administration. Sadegh Shirian, Bakhtiari Moghadam Behnam: Resources. Sadegh Shirian, Bakhtiari Moghadam Behnam: writing–original draft. Norouzi Cholcheh Morteza, Saeed Habibin Dehkordi, Sadegh Shirian, Bakhtiari Moghadam Behnam: writing–review and editing.

Ethics Statement

The Ethical Committee of Shahre‐Kord University approval this study (Approval number: IR.SKU.REC.1404.014). All the experiments were done in accordance with the rights of the animals, and the animals were not harmed in the clinical trial.

Conflicts of Interest

The authors declare no conflicts of interest.

Peer Review

The peer review history for this article is available at https://www.webofscience.com/api/gateway/wos/peer‐review/10.1002/vms3.70578.

Shirian, S. , Cholcheh M. N., Dehkordi S. H., and Moghadam B. B.. 2025. “Therapeutic Effect of Barberry (Berberis vulgaris) Root Extract on Experimental Cutaneous Leishmaniosis in a BALB/c Mouse Model.” Veterinary Medicine and Science 11, no. 5: 11, e70578. 10.1002/vms3.70578

Funding: This study did not receive funding.

Data Availability Statement

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Data Availability Statement

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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