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Effects of Senegenin against hypoxia/reoxygenation-induced injury in PC12 cells

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Chinese Journal of Integrative Medicine Aims and scope Submit manuscript

Abstract

Objective

To investigate the effect and the potential mechanism of Senegenin (Sen) against injury induced by hypoxia/reoxygenation (H/R) in highly differentiated PC12 cells.

Methods

The cultured PC12 cells were treated with H/R in the presence or absence of Sen (60 μmol/L). Four groups were included in the experiment: control group, H/R group, H/R+Sen group and Sen group. Cell viability of each group and the level of lactate dehydrogenase (LDH) in culture medium were detected for the pharmacological effect of Sen. Hoechst 33258 staining and annexin V/propidium iodide double staining were used to analyze the apoptosis rate. Moreover, mitochondrial membrane potential (△Ψm), reactive oxygen species (ROS) and intracellular free calcium ([Ca2+]i) were measured by fluorescent staining and flow cytometry. Cleaved caspase-3 and activity of NADPH oxidase (NOX) were determined by colorimetric protease assay and enzyme linked immunosorbent assay, respectively.

Results

Sen significantly elevated cell viability (P<0.05), decreased the leakage of LDH (P<0.05) and apoptosis rate (P<0.05) in H/R-injured PC12 cells. Sen maintained the value of △Ψm (P<0.05) and suppressed the activity of caspase-3 (P<0.05). Moreover, Sen reduced ROS accumulation P<0.05) and [Ca2+]i increment (P<0.05) by inhibiting the activity of NOX (P<0.05).

Conclusion

Sen may exert cytoprotection against H/R injury by decreasing the levels of intracellular ROS and [Ca2+]i, thereby suppressing the mitochondrial pathway of cellular apoptosis.

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Authors and Affiliations

  1. Department of Pathophysiology, Key Laboratory of State Administration of Traditional Chinese Medicine of the People’s Republic of China, School of Medicine, Jinan University, Guangzhou, 510632, China

    Xiao-qing Zhu  (朱小青), Xue-min Li  (李学敏), Yan-dong Zhao  (赵严冬), Xi-luan Ji  (纪惜銮), Yan-ping Wang  (王彦平), Yong-mei Fu  (付咏梅), Hua-dong Wang  (王华东), Da-xiang Lu  (陆大祥) & Ren-bin Qi  (戚仁斌)

  2. Clifford Hospital, Guangzhou University of Chinese Medicine, Guangzhou, 511496, China

    Xiao-qing Zhu  (朱小青)

  3. High Magnetic Field Laboratory, Chinese Academy of Sciences, Hefei, 230031, China

    Xue-min Li  (李学敏)

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  1. Xiao-qing Zhu  (朱小青)
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  8. Da-xiang Lu  (陆大祥)
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Corresponding author

Correspondence to Ren-bin Qi  (戚仁斌).

Additional information

Supported by the Natural Science Foundation of Guangdong Province, China (No. 2014A030313394), the Project of Science and Technology of Guangzhou, China (No. 2014J4100098), the National Program on Key Basic Research Project (973 Program, No. 2011CB707501), and the Fundamental Research Funds for the Central Universities in China (No. 21613401)

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Zhu, Xq., Li, Xm., Zhao, Yd. et al. Effects of Senegenin against hypoxia/reoxygenation-induced injury in PC12 cells. Chin. J. Integr. Med. 22, 353–361 (2016). https://doi.org/10.1007/s11655-015-2091-8

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  • DOI: https://doi.org/10.1007/s11655-015-2091-8

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