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Molecular & Cellular Toxicology

, Volume 15, Issue 2, pp 209–219 | Cite as

Andrographolide induce human embryonic stem cell apoptosis by oxidative stress response

  • Huarong Huang
  • Huanhuan Cao
  • Chun Xing
  • Yunfen Hua
  • Ming ZhangEmail author
  • Lifang JinEmail author
Original Paper
  • 8 Downloads

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.

Keywords

Andrographolide Human embryonic stem cells Apoptosis Oxidative stress Nrf2 pathway 

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Copyright information

© The Korean Society of Toxicogenomics and Toxicoproteomics and Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.College of Life and Environmental ScienceHangzhou Normal UniversityHangzhouChina
  2. 2.College of Life ScienceShaoxing UniversityShaoxing, ZhejiangChina
  3. 3.Hangzhou Precision Medicine Research CenterHangzhouChina
  4. 4.Hangzhou Economic and Technology Development Area Biomedical Public PlatformHangzhouChina
  5. 5.College of pharmacy pharmaceutical scienceZhejiang University of TechnologyHangzhouChina
  6. 6.College of Life ScienceZhejiang UniversityHangzhouChina

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