Probing the Potential Mechanism of Quercetin and Kaempferol against Heat Stress-Induced Sertoli Cell Injury: Through Integrating Network Pharmacology and Experimental Validation

Abstract

Quercetin and kaempferol are flavonoids widely present in fruits, vegetables, and medicinal plants. They have attracted much attention due to their antioxidant, anti-inflammatory, anticancer, antibacterial, and neuroprotective properties. As the guarantee cells in direct contact with germ cells, Sertoli cells exert the role of support, nutrition, and protection in spermatogenesis. In the current study, network pharmacology was used to explore the targets and signaling pathways of quercetin and kaempferol in treating spermatogenic disorders. In vitro experiments were integrated to verify the results of quercetin and kaempferol against heat stress-induced Sertoli cell injury. The online platform was used to analyze the GO biological pathway and KEGG pathway. The results of the network pharmacology showed that quercetin and kaempferol intervention in spermatogenesis disorders were mostly targeting the oxidative response to oxidative stress, the ROS metabolic process and the NFκB pathway. The results of the cell experiment showed that Quercetin and kaempferol can prevent the decline of cell viability induced by heat stress, reduce the expression levels of HSP70 and ROS in Sertoli cells, reduce p-NF-κB-p65 and p-IκB levels, up-regulate the expression of occludin, vimentin and F-actin in Sertoli cells, and protect cell structure. Our research is the first to demonstrate that quercetin and kaempferol may exert effects in resisting the injury of cell viability and structure under heat stress.

Antioxidant, anti-inflammatory, and anti-reprotoxic effects of kaempferol and vitamin E on lead acetate-induced testicular toxicity in male rats

“…The kaempferol exhibited the antioxidant properties to improve the Leydig cells membranes and facilitate cholesterol transfer into mitochondria ( Alam et al , 2020 ), subsequently improving steroidogenesis as StAR protein levels and CYP11A1, 3β-HSD, and 17β-HSD activities and testosterone concentration were upregulated ( Liu et al , 2022 ) as reported in the current research which illustrated that kaempferol administration caused testicular upregulation 3β-HSD, 17β-HSD, and CTP11A1 enzymes and StAR protein gene expression which positively effect on testosterone levels in rats cotreated with kaempferol. Furthermore, the administration of vitamin E following exposure to lead acetate caused elevation of testicular 13β-and 17β- HSD activities and StAR levels due to the ability of vitamin E to promote gene expression of StAR protein and mRNA steroidogenic enzyme in Leydig cells that can upregulate testosterone concentration ( Malmir et al , 2021 ).…”

“…Importantly, the effects of kaempferol on the reproductive system have not yet been fully reported. Kaempferol, like other flavonoids that affect LH-and FSH-secreting gonadotroph cells by incrementing their secretory granules ( Zhang et al , 2023 ), which results from the ability of kaempferol to upregulate hypophyseal GnRHr , FSHβ , and LHβ genes due to kaempferol’s antioxidant, antiapoptotic, and anti-inflammatory effects ( Liu et al , 2022 ). In addition, kaempferol has the ability to upregulate the testicular receptor expression of LH and FSH due to its powerful antioxidant properties that protect Leydig and Sertoli cells from Pb oxidative damage ( Lee et al , 2023 ), which promotes testicular steroidogenic enzyme expression which enhanced testosterone biosynthesis, as well as improvement the count, and viability of sperm ( Kluska et al , 2022 , Liu et al , 2022 ).…”

“…These alterations were improved and enhanced by administering kaempferol and vitamin E due to their antioxidant and anti-inflammatory properties ( Sukmawati et al , 2019 , Liu et al , 2022 ). In addition, the administration of kaempferol in the current experiment exhibits potent antioxidant and anti-inflammatory activities in testicular tissues via reducing the production of pro-inflammatory mediators and oxidative stress, which can reduce edema and degenerative changes in testicular tissues, protect DNA from oxidative stress ( Akefe et al , 2020 ), enhance the regeneration process in the testicular tissues, seminiferous tubules, Sertoli and Leydig cells as in ( Fig.…”

“…Kaempferol, ″3,5,7-trihydroxy-2-(4-hydroxyphenyl) -4H- chromen-4-one″ is a type of flavonoids, a polyphenol and lipophilic, also known as kaempferol flavonol, kaempferide, kaempferol-3 ( Jin et al , 2023 ), it can be extracted from tea and many vegetables and fruits such as green cabbage, broccoli, olive oil, apple, chives, cucumber, strawberries, lettuce, onion, grapes, spinach, tomatoes, horseradish, and cowpea ( Alam et al , 2020 ). Studies in vivo and in vitro reported that kaempferol possesses therapeutic effects in neurodegenerative disease, colitis, and in acute and chronic inflammatory disease, including acute lung injury and postmenopausal bone loss ( Ren et al , 2019 , Liu et al , 2022 , Lim et al , 2023 ), as well as, kaempferol has a protective role in liver injury and prevents metabolic disorders, furthermore, anticancer role in breast cancer, and hepatocellular carcinoma ( Wang et al , 2019 , Sharma et al , 2021 , Fan et al , 2023 ).…”

“…Recently, Liu et al (2022) reported that kaempferol exerts anti-inflammatory and antioxidant effects against heat stress-induced Sertoli cell injury in vitro, which prevents spermatogenic disorders; however, the protective effects and mechanisms of kaempferol alone and with vitamin E on lead acetate-induced hypothalamic-hypophyseal-testicular dysfunction in vivo have not been reported. Based on this limitation, the present research aimed to investigate the possible mechanism(s) by which kaempferol, separately or combined with vitamin E, improves testicular function in rats exposed to lead acetate.…”