Monitoring the Nanocurcumin Effect on Lead Exposure in the Huh۷-۱x-ARE-luc Cell Line

Objective: The human genome consists of protective genes, which contain a sequence known as the antioxidant responsive element (ARE) located in their promoter regions. ARE is specific to the transcription factor NF-E۲ related factor۲ (Nrf۲). This signaling pathway is the major defense mechanism against oxidative stress that comprises the chemoprotective response. The cell line that expresses the ARE reporter is sensitive for the detection of ARE activating compounds. It can help to identify toxicity risk and antioxidant activity of chemicals and drugs. Methods: We used a stable Huh۷ ARE-reporter cell line in this study. Metabolic activity of these cells in different concentrations of lead (۰ to ۸۰ micromole) was evaluated by the MTT test. We assessed the effects of oxidative stress. We exposed the Huh۷ ARE-reporter cell line to different concentrations of lead, an oxidative stress inducer, and nanocurcumin, an antioxidant compound, after which we investivated luciferase activity. Real-time PCR was used to detected AREKEAP۱ pathway gene expression. Result: Lead, at ۳۰ μM, suppressed ۵۰% of the cells’ metabolic activity.Cells treated with both lead (۳۰ μM) and nanocurcumin at ۴ μM and ۸ μM had decreased luciferase activity compared to those treated with only lead. This activity increased when we increased the nanocurcumin concentration to ۱۶ μM. Real-time PCR analysis showed decreased NQO۱ and NRF۲ expressions in cells treated with both lead (۳۰ μM) and nanocurcumin (۴ μM and ۸ μM) compared to just lead treated cells. However, NQO۱ and NRF۲ had increased expressions in cells treated with both lead (۳۰ μM) and nanocurcumin (۱۶ μM) compared to just lead treated cells. Conclusion: Nano curcumin, as an antioxidant, significantly reduced the toxic effects of lead toxic. This effect probably occurred through the AREKEAP۱ pathway. Hence, nanocurcumin could be used as an antioxidant to reduce oxidative stress.

کلیدواژه ها:

Nrf۲ ، Antioxidant responsive element (ARE) ، Oxidative stress ، Reporter cell line ، Nanocurcumin ، عوامل پاسخگو به آنتی اکسیدانها ، استرس اکسیداتیو ، رده سلولی گزارشگر ، نانوکورکومین