Highlights
- •A high-fat diet induces obesity, oxidative stress and sperm deformities, altering testis physiology.
- •A high-fat diet alters testicular hormonal balance and the maintenance of sperm quality.
- •Orlistat treatment in obese rats revealed substantial testicular damage and aggravated the effects of a high-fat diet.
- •Artemisia Annua Extract (AAE) counteracts high-fat diet deleterious effect on testicular functions.
- •AAE ameliorates high-fat diet-induced spermatogenesis malformations and hormonal secretion.
- •AAE can be used to recover the hormonal balance and improve sperm quality in men with obese fertility disorders.
Abstract
Ethnopharmacological relevance
Artemisia annua L., known as “sweet wormwood,” is widely used in Egyptian folk medicine. Egyptians implement the aerial parts in the treatment of respiratory, digestive and sexual dysfunctions. However, the mechanism by which Artemisia annua improves testicular function is still being discovered.
Aim of the study
This study aimed to evaluate the modulatory effects of the crude leaf extract of Artemisia annua (AAE) on a high-fat diet induced testicular dysfunction in rats and compare it with the antilipolytic drug Orlistat.
Material and methods
Forty adult rats were randomly classified and assigned to four groups. The first group typically consumed a balanced diet and served as a negative control (GP1). A high-fat diet-induced obesity was applied to the other three groups for 12 weeks. A positive control remained on HFD for another 8 weeks, which is GP2. Other groups were administered for 8 consecutive weeks either with Orlistat (50 mg/kg body weight) or AAE (100 mg/kg body weight), which have been defined as GP3 and GP4, respectively. Testosterone (TST), follicle-stimulating hormone (FSH) and luteinizing hormone (LH) were determined in the sera of all groups. In addition, the oxidant/antioxidant biomarkers such as protein carbonyl, malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) activities, lactate dehydrogenase (LDH) and creatine kinase isoenzyme-B (CK-MB) were determined. An immunohistochemical stain with the apoptotic marker caspase-3 and the proliferating cell nuclear antigen (PCNA) were also investigated.
Results
In the testes of the obese group, the results showed hormonal imbalance, an increase in oxidative stress biomarkers and apoptosis. In the group treated with orlistat (GP3), noticeably more perturbations were noted. The obese rats that had been treated with AAE (GP4) showed a significantly reduced level of oxidative stress, hormonal balance restoration and reduced apoptosis.
Conclusions
The crude leaf extract of A. annua is a potential herbal therapeutic for the treatment of obesity-related testicular dysfunction and the restoration of hormonal imbalance in obese rats.
Graphical abstract
Introduction
Obesity is a major health problem worldwide with a high incidence among children and adults (World Health Organization, 2021). Four billion people in the global population would be overweight or obese in 2025; approximately 16% of them would be obese (Bodirsky et al., 2020). Obese individuals are at significant risk of diabetes type 2 (T2-DM), coronary heart diseases, male obesity-related secondary hypogonadism and cancer (Fernandez et al., 2019; Sarma et al., 2021). Particularly in men, obesity is strictly linked to elevated plasma levels of leptin, proinflammatory markers and excess body fatness (Obradovic et al., 2021). This pathophysiological condition deteriorates infertility, sexual ability and muscle mass (De Lorenzo et al., 2018).
Obesity increases testosterone conversion into estradiol, reduces luteinizing hormone secretion and eventually hinders the synthesis and production of testosterone (TST) from Leydig cells. Obesity-associated hypotestosteronemia is a reversible status and weight loss can restore total TST levels (Di Vincenzo et al., 2018). Obesity has been found to progressively worsen the sperm count and motility. Increase the production of reactive oxygen species (ROS), increase apoptosis and increase the risk of azoospermia or oligozoospermia (Sultan et al., 2020).
Orlistat is tetrahydrolipstatin, an endogenous lipstatin isolated from Streptomyces toxytricini and widely used in the management of obesity (Bansal and Al Khalili, 2020). It acts via inhibition of the gastric and pancreatic lipases that prevent hydrolysis of the long-chain triglycerides (Tada et al., 2020). Inhibition of lipolysis reduces AMP-activated protein kinase (AMPK) activation (Gauthier et al., 2008) and the generation of CD8(+) memory T cells (O’Sullivan et al., 2014).
Medicinal plants are an excellent approach for treating obesity because they are a safer alternative to chemical drugs in the long run (Chung et al., 2021; Hassan et al., 2022). Artemisia spp., one of the medicinal plants, belongs to the family Asteraceae (Darqui et al., 2021). There are over 500 species in the genus Artemisia L., which is predominantly found in the northern temperate regions of the world (Abad et al., 2013). A. annua L. is abundantly distributed in Asia, Europe and North America. This fragrant plant, also known as sweet wormwood, was used in traditional medicine. A. annua L. was utilized to isolate artemisinin, a sesquiterpene lactone with antimalarial properties. Numerous phytochemicals and essential oils (EOs) with intriguing antibacterial properties have been discovered and isolated from A. annua (Msaada et al., 2015).
Over 100 compounds were identified in this plant. Artemisia ketone and oxygenated monoterpenes made up the majority of the essential oils (30.7%), as well as camphor. Minimum inhibitory concentrations (MICs) of Artemisia camphorata L. were found to be effective against enterotoxigenic, enteropathogenic and enteroinvasive E. coli (Zhang et al., 2017). A. annua has been used as tonics, antimalarials, anthelmintics, antidiabetics and for bronchitis, ulcers and the treatment of tuberculosis (Shishehgar et al., 2021). Previous study suggested that AAE could suppress obesity and prevent fatty liver by moderating differentiation of adipocytes and fat accumulation in vitro and in vivo (Choi et al., 2021). Additionally, obese mice treated with AAE showed significantly less weight gain, with no substantial changes in food intake volume or calories.
This investigation aims to assess the modulatory effects of the crude leaf extract of A. annua(AAE) on rat testicular dysfunction driven by a high-fat diet, in addition to comparing it with that of the anti-lipolytic pill Orlistat.
Section snippets
Chemicals
Orlistat 120 mg capsules were purchased from October Pharmaceutical Co., Egypt. Phosphate buffered saline (PBS), bovine serum albumin (BSA), cleavage caspase-3 and proliferating cell nuclear antigens (PCNA) antibodies were purchased from Sigma Aldrich Company (Berlin, Germany). Testosterone (TST) MAIA® kit was purchased from Biochem Immuno System Company (place, country). FSH ELISA kit was purchased from MYBIOSOURCE Company (MBS2021901), Rat LH (luteinizing hormone) ELISA Kit (E-EL-R0026) was
Phytochemical composition of A. annua leaf extract
The extract revealed a rich content of flavonoids and saponins and moderate anthocyanin and phenolic contents, as shown in Table 1. Moreover, the in vitro free radical scavenging activity shows enhanced antioxidant properties.
Gas chromatography-mass spectroscopy profiling of A. annua
A total of 12 compounds were identified from the GC-MS analysis of the ethanol fraction of A. annua leaves, as shown in Fig. 2. The chemical constituents with their retention time (RT), molecular formula, molecular weight (MW) and concentration (%) are presented in Table 2
Discussion
A high-fat diet (HFD) or western diet is full of calories; consuming the HFD for a prolonged period of time causes an obesity-like phenotype in animal models. These phenotypes include increased weight gain, the expansion of fat tissue, and changes to the immune and physiological systems (Abdelsadik, 2018; Bari et al., 2020). Male obesity in general causes insulin resistance, increases low-grade inflammation, and obstructs testicular dysfunction (Kunz et al., 2021; Püschel et al., 2022). When
Ethical approval
This study was carried out according to the guidelines approved by the Research Ethical Committee, Faculty of Science, Tanta University, Egypt (IACUC–SCI–TU-0217).
Funding
This research received no funding.
CRediT authorship contribution statement
Samer A. El-Sawy: Conceptualization, Data curation, and, Formal analysis, Formal analysis of the manuscript. Yahia A. Amin: Supervision, Validation, Formal analysis of the manuscript, Data curation, Writing – original draft, Writing – review & editing, Writing review and editing the manuscript. Sabry A. El-Naggar: Methodology, and, Writing – original draft, Writing original draft of the manuscript. Ahmed Abdelsadik: Project administration, Software, Visualization, Formal analysis, Formal
Declaration of competing interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
