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Andrographolide induce human embryonic stem cell apoptosis by oxidative stress response

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Abstract

Backgrounds

The anti-inflammatory effect of andrographolide is widely accepted; however, its exact role in reproductive toxicity requires further elucidation. The embryonic stem cell test (EST) is a promising system for detecting the reproductive toxicity of drugs in vitro. In this study, we applied a prediction model to our EST data after classifying andrographolide according to published criteria. The possible mechanism of andrographolide reproductive toxicity was also studied.

Methods

Reproductive toxicity of andrographolide was evaluated in vitro EST model and in vivo mouse model. Human embryonic stem cells (ESCs) were cultured with different concentrations of andrographolide with or without N-acetyl-L-cysteine (NAC). Cell viability was assessed with MTT assay, and reactive oxygen species (ROS) level was measured with DCFH-DA assay. Gene and protein expression levels were measured with qRT-PCR and western-blot, respectively.

Results

Results showed that andrographolide exhibited strong reproductive toxicity according to the prediction model of the EST and mouse studies. An increase in ROS levels, damage to mitochondrial membrane potential, and induction of caspase-3 were observed in the andrographolide-treated human ESCs. Scavenging of andrographolide-induced ROS by NAC blocked these activities. Western blot and qRT-PCR analysis revealed that the nuclear factor erythroid-2-related factor 2 (Nrf2) protein and its target antioxidant genes were up-regulated after andrographolide treatment at certain concentrations. Furthermore, NAC treatment significantly increased the activity of the Nrf2 signaling pathway.

Conclusion

We demonstrated that andrographolide is a drug with strong reproductive toxicity, which resulted from ROS-mediated oxidative stress. In addition, the Nrf2 pathway appears to be involved in the NAC protection of human ESCs against andrographolide-induced cell apoptosis.

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Authors and Affiliations

  1. College of Life and Environmental Science, Hangzhou Normal University, Hangzhou, China

    Huarong Huang & Chun Xing

  2. College of Life Science, Shaoxing University, Shaoxing, Zhejiang, China

    Lifang Jin

  3. Hangzhou Precision Medicine Research Center, Hangzhou, China

    Huarong Huang, Huanhuan Cao, Yunfen Hua, Ming Zhang & Lifang Jin

  4. Hangzhou Economic and Technology Development Area Biomedical Public Platform, Hangzhou, China

    Huanhuan Cao

  5. College of pharmacy pharmaceutical science, Zhejiang University of Technology, Hangzhou, China

    Yunfen Hua

  6. College of Life Science, Zhejiang University, Hangzhou, China

    Ming Zhang

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  1. Huarong Huang
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  2. Huanhuan Cao
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Correspondence to Ming Zhang or Lifang Jin.

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Huang, H., Cao, H., Xing, C. et al. Andrographolide induce human embryonic stem cell apoptosis by oxidative stress response. Mol. Cell. Toxicol. 15, 209–219 (2019). https://doi.org/10.1007/s13273-019-0024-x

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  • DOI: https://doi.org/10.1007/s13273-019-0024-x

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