Promising anti- cervical carcinoma and inflammatory agent, Resveratrol targets poly (ADP-ribose) polymerase 1 (PARP-1) induced premature ovarian failure with a potent enzymatic modulatory activity

Highlights

• We report resveratrol as a potential target for PARP-1 and its modulator from a high-throughput virtual screening method.

• Resveratrol strongly inhibited HT-3 cell proliferation, the IC₅₀ value is 0.65 μM.

• The compound induced HT-3 cell apoptosis in a dose-response manner.

• Resveratrol restored ovarian function through increasing AMH levels, and diminishing ovarian inflammation.

Abstract

Radioprotective effects of Resveratrol is well known in normal cells exposed to the damaging effects of ionizing radiation however, its potential radioprotective effect on ovarian follicle formation and development is still uncertain. Astonishingly, it has been reported that PARP contributed to the pathogenesis of immune-mediated ovarian injury. In this paper, Resveratrol was tested for its inflammatory, anti-cervical carcinoma activity, and checked its targets poly (ADP-ribose) polymerase 1 (PARP-1) induced premature ovarian failure with a potent enzymatic modulatory activity. Through high-throughput virtual screening method, Resveratrol was screened to find its target. That the compound strongly inhibited cervical carcinoma HT-3 cell. The cell proliferation was evaluated by an CCK-8 assay, and the cell apoptosis was assessed by a flow cytometry. Rat model of premature ovarian failure was used to introduce resveratrol preparation and rtPCR was done to measure expression of apoptosis related markers. We report resveratrol as a potential target for PARP-1 and its modulator from a high-throughput virtual screening method. Resveratrol was measured its anti-cervical carcinoma activity by using an CCK-8 assay, which suggested that the compound strongly inhibited HT-3 cell proliferation, the IC₅₀ value is 0.65 μM. In addition, the compound induced HT-3 cell apoptosis in a dose-response manner. Resveratrol preserves the entire ovarian follicle pool manifested by increasing serum anti-Müllerian hormone (AMH) levels. Study suggest that resveratrol restored ovarian function through increasing AMH levels, and diminishing ovarian inflammation, predominantly modulation of PPAR-1 and inhibition of inflammatory cytokines. Resveratrol was identified targets for PARP-1 from a high-throughput virtual screening method, strongly inhibited PARP-1 protein and HT-3 cell proliferation. Resveratrol is a promising PARP-1 modulator with anti-cervical carcinoma activity, which deserves further investigation.

Introduction

Cervical cancer is the second largest malignancy in women and a major cause of cancer-related deaths around the world and has the highest mortality rate among all gynecologic tumors (Moore et al., 2010; Kitagawa et al., 2015). The main treatment strategy of cervical cancer includes surgery, radiotherapy, and drug therapy, but conventional chemotherapy has a limited therapeutic effect and causes serious systemic toxicity (Kitagawa et al., 2015; Kumar and Gupta, 2016). The initial introduction of drug therapy in the field of cervical cancer treatment is aimed at patients with relapse and metastasis who failed in the first treatment (Loizzi et al., 2008, Adriaens et al., 2009) modulator. However, the tolerance of tumor cells to radiotherapy gradually enhances with the extension of treatment time (Loizzi et al., 2008, Adriaens et al., 2009, Kumar and Gupta, 2016), which caused ovarian damage and accelerates menopause may later ends up with permanent infertility (Adriaens et al., 2009). Occasionally high doseexposure leads to sterilization, but in certain cases premature ovarian failure is outcome due to partial depletion of the primordial follicle (Gosden et al., 1997; Kim et al., 2014). Therefore, to protect ovarian function and fertility of cervical cancer survivors need to be considered, and new strategy to be identified.

Poly ADP ribose polymerase-1 (PARP-1), a well-studied member of the nuclear transcription factors receptor superfamily and cellular ribozyme which catalyzes the ribosylation of poly (ADP) and have a role in the inflammatory responses control (Vickers, 2017; Miwa and Masutani, 2007) in eukaryotic cells. PARP is involved in the process of DNA repair and replication and plays an important role in repairing and maintaining gene integrity after DNA damage (Soldatenkov and Smulson, 2000; Bryant et al., 2005), and if it could not be repaired or wrongly repaired, it will lead to cell mutation, which may eventually lead to the occurrence of tumor (Rodríguez et al., 2015). In recent years, research results show that PARP is also related to development of a variety of diseases, including many inflammatory diseases, autoimmune diseases and occurrence and development of tumor (Wong et al., 2010; Morita et al., 2012; Wang et al., 2014). PPAR-c regulates the synthesis of steroid hormones in the granulosa cells (Delerive et al., 2001), and the disruption of PPAR-c in the ovary leads to female subfertility (Huang et al., 2008). Nevertheless, granulosa cell specific deletion of PPAR results in marked impairment of ovulation due to defective follicular rupture (Cui et al., 2002). Intriguingly, following ionizing radiation exposure, intestinal inflammatory processes have been recorded with a dramatic impairment of the PPAR-c level (Kim et al., 2008). Numerous in vitro studies have authenticated the radioprotective effects of Resveratrol in normal cells exposed to the damaging effects of ionizing radiation (Linard et al., 2008; Moding et al., 2013) however, its potential radioprotective effect on ovarian follicle formation and development is still uncertain. Astonishingly, it has been reported that PARP contributed to the pathogenesis of immune-mediated ovarian injury (Vilar et al., 2011). In this paper, Resveratrol was identified PARP-1 protein as its target from a high-throughput virtual screening method, which was then measured for its role in cervical carcinoma.