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Ginsenoside Rb1 Ameliorates Autophagy of Hypoxia Cardiomyocytes from Neonatal Rats via AMP-Activated Protein Kinase Pathway

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Abstract

Objective

To investigate whether ginsenoside-Rb1 (Gs-Rb1) improves the CoCl-induced autophagy of cardiomyocytes via upregulation of adenosine 5′-monophosphate-activated protein kinase (AMPK) pathway.

Methods

Ventricles from 1- to 3-day-old Wistar rats were sequentially digested, separated and incubated in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum for 3 days followed by synchronization. Neonatal rat cardiomyocytes were randomly divided into 7 groups: control group (normal level oxygen), hypoxia group (500 μmol/L CoCl2), Gs-Rb1 group (200 μmol/L Gs-Rb1 + 500 μmol/L CoCl2), Ara A group (500 μmol/L Ara A + 500 μmol/L CoCl2), Ara A+ Gs-Rb1 group (500 μmol/L Ara A + 200 μmol/L Gs-Rb1 + 500 μmol/L CoCl2), AICAR group [1 mmol/L 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) + 500 μmol/L CoCl2], and AICAR+Gs-Rb1 group (1 mmol/L AICAR + 200 μmol/L Gs-Rb1 + 500 μmol/L CoCl2). Cells were treated for 12 h and cell viability was determined by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay and cardiac troponin I (cTnI) levels were detected by enzyme-linked immunosorbent assay (ELISA). AMPK activity was assessed by 2′,7′-dichlorofluorescein diacetate (DCFH-DA) ELISA assay. The protein expressions of Atg4B, Atg5, Atg6, Atg7, microtubule-associated protein 1A/1B-light chain 3 (LC3), P62, and active-cathepsin B were measured by Western blot.

Results

Gs-Rb1 significantly improved the cell viability of hypoxia cardiomyocytes (P<0.01). However, the viability of hypoxia-treated cardiomyocytes was significantly inhibited by Ara A (P<0.01). Gs-Rb1 increased the AMPK activity of hypoxia-treated cardiomyocytes. The AMPK activity of hypoxia-treated cadiomyocytes was inhibited by Ara A (P<0.01) and was not affected by AICAR =0.983). Gs-Rb1 up-regulated Atg4B, Atg5, Beclin-1, Atg7, LC3B II, the LC3B II/I ratio and cathepsin B activity of hypoxia cardiomyocytes (P<0.05), each of these protein levels was significantly enhanced by Ara A (all P<0.01), but was not affected by AICAR (all P>0.05). Gs-Rb1 significantly down-regulated P62 levels of hypoxic cardiomyocytes (P<0.05). The P62 levels of hypoxic cardiomyocytes were inhibited by Ara A (P<0.05) and were not affected by AICAR (P=0.871).

Conclusion

Gs-Rb1 may improve the viability of hypoxia cardiomyocytes by ameliorating cell autophagy via the upregulation of AMPK pathway.

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Authors and Affiliations

  1. Department of Cardiology, the People’s Hospital of China Medical University, the People’s Hospital of Liaoning Province, Shenyang, 110016, China

    Sheng-nan Dai, Ai-jie Hou, Xiao-ming Chen, Hua-ting Huang & Hong-liang Kong

  2. International Education College, Shenyang Normal University, Shenyang, 110034, China

    Shu-mei Zhao

  3. Department of Cardiology, the First Affiliated Hospital of Dalian Medical University, Dalian, 116044, China

    Bo-han Chen

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  1. Sheng-nan Dai
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  2. Ai-jie Hou
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  3. Shu-mei Zhao
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  4. Xiao-ming Chen
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  5. Hua-ting Huang
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  6. Bo-han Chen
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  7. Hong-liang Kong
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Correspondence to Hong-liang Kong.

Additional information

Supported by Shenyang Innovation Foundation of Science and Technology-the Application Projects of Basic Research (No. F15-199-1-06), Liaoning Province Science and Technique Foundation of China (No. 2015020282)

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Dai, Sn., Hou, Aj., Zhao, Sm. et al. Ginsenoside Rb1 Ameliorates Autophagy of Hypoxia Cardiomyocytes from Neonatal Rats via AMP-Activated Protein Kinase Pathway. Chin. J. Integr. Med. 25, 521–528 (2019). https://doi.org/10.1007/s11655-018-3018-y

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  • DOI: https://doi.org/10.1007/s11655-018-3018-y

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