Keap1/Nrf2-independent antioxidative activity of Phyllanthus amarus extract in zebrafish

Vu Thanh Nguyen, Nguyen Thi Tam Thanh, Nguyen Phuong Phi, Duong Nguyen Mai Que, Luu Thu Hien, Le Luu Phuong Hanh, Ngo Huynh Phuong Thao, Tong Xuan Nguyen, Pham Thanh Luu, Nguyen Hoang Thuy Vy, Dinh Thi Thuy
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Authors

  • Vu Thanh Nguyen Division of Aquacultural Biotechnology, Biotechnology Center of Ho Chi Minh City, 2374 Highway 1, Quarter 2, Trung My Tay Ward, District 12, Ho Chi Minh City, Vietnam https://orcid.org/0000-0002-3458-8813
  • Nguyen Thi Tam Thanh Division of Aquacultural Biotechnology, Biotechnology Center of Ho Chi Minh City, 2374 Highway 1, Quarter 2, Trung My Tay Ward, District 12, Ho Chi Minh City, Vietnam
  • Nguyen Phuong Phi Division of Aquacultural Biotechnology, Biotechnology Center of Ho Chi Minh City, 2374 Highway 1, Quarter 2, Trung My Tay Ward, District 12, Ho Chi Minh City, Vietnam
  • Duong Nguyen Mai Que Division of Aquacultural Biotechnology, Biotechnology Center of Ho Chi Minh City, 2374 Highway 1, Quarter 2, Trung My Tay Ward, District 12, Ho Chi Minh City, Vietnam
  • Luu Thu Hien Division of Aquacultural Biotechnology, Biotechnology Center of Ho Chi Minh City, 2374 Highway 1, Quarter 2, Trung My Tay Ward, District 12, Ho Chi Minh City, Vietnam
  • Le Luu Phuong Hanh Division of Aquacultural Biotechnology, Biotechnology Center of Ho Chi Minh City, 2374 Highway 1, Quarter 2, Trung My Tay Ward, District 12, Ho Chi Minh City, Vietnam
  • Ngo Huynh Phuong Thao Division of Aquacultural Biotechnology, Biotechnology Center of Ho Chi Minh City, 2374 Highway 1, Quarter 2, Trung My Tay Ward, District 12, Ho Chi Minh City, Vietnam
  • Tong Xuan Nguyen Institute of Environmental Science, Engineering, and Management, Industrial University of Ho Chi Minh City, 12 Nguyen Van Bao Street, Ward 4, Go Vap District, Ho Chi Minh City, Vietnam
  • Pham Thanh Luu Institute of Tropical Biology, Vietnam Academy of Science and Technology, 9/621 Hanoi Highway, Linh Trung Ward, Thu Duc District, Ho Chi Minh City, Vietnam
  • Nguyen Hoang Thuy Vy Division of Aquacultural Biotechnology, Biotechnology Center of Ho Chi Minh City, 2374 Highway 1, Quarter 2, Trung My Tay Ward, District 12, Ho Chi Minh City, Vietnam
  • Dinh Thi Thuy Department of Engineering and Technology, Van Hien University, 665-667-669 Dien Bien Phu Street, Ward 1, District 3, Ho Chi Minh City, Vietnam

DOI:

https://doi.org/10.15625/1811-4989/17475

Abstract

The Keap1 protein (Kelch-like ECH-related protein 1) and the Nrf2 transcription factor (NF-E2-related factor 2) are important systems for maintaining homeostasis, redox, and metabolism. Based on the Keap1/Nrf2 pathway, the antioxidative mechanism of P. amarus extract (PAE) was predicted. In this paper, we evaluated the protective effects of PAE on the oxidative toxicity induced by sodium arsenite (NaAsO2) and hydrogen peroxide (H2O2) in zebrafish larvae. We first determined that the LC50 values for NaAsO2, H2O2, and PAE at 3.5 days postfertilization (dpf) were 1 mM, 3 mM, and 200 μg/mL, respectively. Then, to assess the antioxidant effects of P. amarus, 3.5 dpf zebrafish larvae were pretreated with PAE at concentrations of 0, 50, 75, and 100 μg/ml for 12 h, and then the PAE solution was replaced with 1 mM NaAsO2 or 3 mM H2O2 to assess challenge survival within 48 h. Interestingly, all three concentrations, 50, 75, and 100 μg/mL PAE, increased the survival rate of zebrafish larvae compared to those larvae exposed to only 1 mM NaAsO2. Similarly, PAE at concentrations of 75 and 100 μg/mL protected zebrafish larvae after exposure to 3 mM H2O2. Real-time qPCR analysis was performed after 3.5 dpf zebrafish larvae were exposed to 100 μg/mL PAE for 12 h to verify whether the increasing antioxidative activity is depended on the Nrf2 pathway. The expression of the Nrf2 target genes glutathione-S-transferase Pi 1 (gstp1) and peroxidase 1 (prdx1) was assessed using real-time qPCR. However, the expression of this gene was not significantly different between control larvae and PAE-treated larvae. Thus, PAE induces antioxidant activity in zebrafish in a Keap1/Nrf2-independent manner.

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Published

28-12-2022

How to Cite

Thanh Nguyen, V., Nguyen, T. T. T., Phuong Phi, N., Mai Que, D. N., Thu Hien, L., Phuong Hanh, L. L., Phuong Thao, N. H., Nguyen, X. T., Thanh Luu, P., Thuy Vy, N. H., & Thi Thuy, D. (2022). Keap1/Nrf2-independent antioxidative activity of Phyllanthus amarus extract in zebrafish. Vietnam Journal of Biotechnology, 20(4), 653–661. https://doi.org/10.15625/1811-4989/17475

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