RESEARCH ARTICLE Ameliorative effects of Saussurea lappa root aqueous extract against Ethephon-induced reproductive toxicity in male rats Mabrouk Attia Abd Eldaim 1,2 | Ehab Tousson 3 | Ibrahim El Tantawy El Sayed 4 | Walaa Mohamed Awd 4 1 Department of Biochemistry and Chemistry of Nutrition, Faculty of Veterinary Medicine, Menoufia University, Egypt 2 Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University, Kyoto, Japan 3 Zoology Department, Faculty of Science, Tanta University, Tanta, Egypt 4 Chemistry Department, Faculty of Science, Menoufia University, Egypt Correspondence Mabrouk Attia Abd Eldaim, Department of Biochemistry and Chemistry of Nutrition, Faculty of Veterinary Medicine, Menoufia University, Sheben Elkom, Egypt. Email: mabroukattia@vet.menofia.edu.eg Abstract This study was designed to evaluate protective effect of Saussurea lappa root aqueous extract against Ethephon (2-chloroethylphosphonic acid)-induced reproductive toxicity in rats. Control group received distilled water. Second group was given S. lappa extract at a dose 50 mg/kg bw. Third group was given Ethephon at a dose 200 mg/kg bw. Fourth, fifth, and sixth groups were given S. lappa extract before, with or after Ethephon administration, respectively. Ethephon intoxication significantly decreased serum levels of follicle stimulating hormone, luteinizing hor- mone, testosterone, and prolactin. Also, it significantly decreased sperms count, vitality, mor- phology index, total motility, progressive motility, and proliferating cell nuclear antigen protein expressions in spermatogonia. However, it significantly increased sperms abnormalities, testicu- lar tissue and DNA damages, P53 protein expressions, noprogressive motility, and immotile sperms. In contrast, S. lappa extract ameliorated these alterations. These results indicated that S. lappa had potential preventive and curative effects against Ethephon-induced reproductive toxicity in rats. KEYWORDS Costus, Ethephon, P53, PCNA, Saussurea lappa, sperm morphometry 1 | INTRODUCTION Plant growth promoters are in continuous use in agriculture. 1,2 Ethephon (2-chloroethylphosphonic acid; Ethrel) is an ethylene-based plant growth regulator used in agriculture. It has direct and indirect effects on agricul- ture and industrial workers. Direct effects occur via inhalation, whereas indirect effects occur through consumption of Ethephon-contaminated fruits and vegetable. 3–5 Ethephon is used for many purposes such as increasing the resistance of cereals to lodging through straw shortening and strengthening, promoting fruits and vegetables maturity, stimulating flowering, side branching and reducing plant height in ornamentals, pro- moting the uniform boll opening, enhancing defoliation, and rubber to increase yield and to reduce bark consumption in cotton. 5,6 Plant growth promoters have been shown to be harmful to liver, kidney, and brain. 7 Ethephon has been reported to have toxic, mutagenic, and teratogenic effects including, gastroenteritis, respira- tory diseases, and necrotic hepatitis, when administered orally, folded retina, microphthalmia, absent tail, and chromosomal aberrations in rats and mice. 8,9 Moreover, it induces biochemical alterations, pancre- atic islet cell tumors in rats, 9,10 glomerulonephritis, glomerulosclerosis, liver fibrosis, stomach necrosis, and increases mammary gland ectasia and ovary stromal cell hyperplasia. 11–13 Many natural products possess significant antioxidant, anti- inflammatory, and anticancer activities which enable them to treat several diseases including testicular toxicity. 14–16 Costus roots (Saus- surea Lappa) are one of these herbal plants. It belongs to the family Asteraceae which is commonly known as sunflower family. Costus is popularly known as Kuth root or Costus and used in various tradi- tional medicines as anti-ulcer, anticonvulsant, anticancer, hepatopro- tective, anti-arthritic, antiviral, and antioxidant 17–20 because it contains many active compounds such as flavonoids, steroids, ter- penes, alkaloids sesquiterpenes, costunolide, dehydrocostus lactone, cynaropicrin, and chlorogenic acid. 21 To the best of our knowledge, studies concerning the toxic effects of Ethephon on the mammalian reproductive function and the Abbreviations: CASA, computer-assisted semen analysis; FSH, follicle- stimulating hormone; LH, luteinizing hormone; P53, tumor protein; PBS, phos- phate-buffered saline; PCNA, proliferating cell nuclear antigen; ROS, reactive oxygen species Received: 19 July 2018 Revised and accepted: 30 September 2018 DOI: 10.1002/tox.22669 Environmental Toxicology. 2018;1–10. wileyonlinelibrary.com/journal/tox © 2018 Wiley Periodicals, Inc. 1