Comparative study of the ameliorating effect of Cyperus esculentus and clomiphene citrate on cadmium-induced toxicity on male reproductive hormones in wistar rats

Abstract

Though herbal remedies have been associated with several functions, the compounds of the herbs and their mechanism of action associated with these functions have always been a task many researchers fail to provide. Such is the case of Cyperus esculentus and its subsequent effect on the reproductive parameters in male animals. This research seeks to ascertain the second messenger signalling pathway through which active compounds of Cyperus esculentus are used to ameliorate toxicity of the reproductive system in rats by assessing hormonal levels of the testis and applying bioinformatics study. Rats of 200–250 g divided into control, cadmium, high-dose cyperus extract (HDCE), low-dose cyperus extract (LDCE), middle-dose cyperus extract (MDCE), and clomid experimental groups (n = 5) were used. All animals were allowed free access to food and water. Except for the control group, others were exposed to a daily 3 g/10 L cadmium chloride solution as their drinking water for the four weeks of administration. HDCE, MDCE, and LDCE received oral Cyperus esculentus ethanolic extract at 3750 mg/kg, 2500 mg/kg, and 1250 mg/kg, respectively, using the previously published LD50 result of 5000 mg/kg. The Clomid group received a solution of Clomid at the standard recommended dose of 2.8 mg/kg. An extract of Cyperus esculentus was assessed for its phytochemical compounds using Gas Chromatography Mass Spectrometry (GCMS) techniques, and compounds with the highest peak area were docked with enzymes of signalling pathways, namely 4UYA (PKC), 4YHJ (PKA), and 5IUZ (PKG). At the end of the administration period, animals were sacrificed in the laboratory using isoflurane anaesthesia. Through cardiac puncture, the serum was extracted for testosterone, FSH, LH, and GnRH assay using ELISA kits. The result of serum testosterone (ng/mL) among the control, cadmium, LDCE, MDCE, HDCE, and Clomid groups in mean ± SD was 1.13 ± 0.02 ng/mL, 0.81 ± 0.08 ng/mL, 0.88 ± 0.09 ng/mL, 0.98 ± 0.10 ng/mL, 1.02 ± 0.07 ng/mL, and 1.09 ± 0.02 ng/mL, respectively. Testosterone levels in the MDCE, HDCE, and Clomid groups were significantly higher than that of Cadmium, whereas those of the MDCE and HDCE were significantly lower than that of the Clomid group at P < 0.05, indicating the ameliorating effect of Clomid and Cyperus esculentus. The same ameliorating results were observed for FSH, LH, and GnRH. A bioinformatics study presented Caryophyllene as the active compound of Cyperus esculentus responsible for its effect following a peak area of 12.47. Furthermore, whereas Clomid showed more affinity and bioactive properties with 4YHJ, caryophyllene binds more with 5IUZ. Therefore, the ameliorating effect of Clomid and Cyperus esculentus through caryophyllene is likely via the cAMP and cGMP second messenger pathways, respectively. Future in vitro studies are required to authenticate these pathways.