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Total Saponins of Panax notoginseng Activate Akt/mTOR Pathway and Exhibit Neuroprotection in vitro and in vivo against Ischemic Damage

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Abstract

Objective

To reveal the neuroprotective effect and the underlying mechanisms of a mixture of the main components of Panax notoginseng saponins (TSPN) on cerebral ischemia-reperfusion injury and oxygen-glucose deprivation/reoxygenation (OGD/R) of cultured cortical neurons.

Methods

The neuroprotective effect of TSPN was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, flow cytometry and live/dead cell assays. The morphology of dendrites was detected by immunofluorescence. Middle cerebral artery occlusion (MCAO) was developed in rats as a model of cerebral ischemia-reperfusion. The neuroprotective effect of TSPN was evaluated by neurological scoring, tail suspension test, 2,3,5-triphenyltetrazolium chloride (TTC) and Nissl stainings. Western blot analysis, immunohistochemistry and immunofluorescence were used to measure the changes in the Akt/mammalian target of rapamycin (mTOR) signaling pathway.

Results

MTT showed that TSPN (50, 25 and 12.5 µ g/mL) protected cortical neurons after OGD/R treatment (P<0.01 or P<0.05). Flow cytometry and live/dead cell assays indicated that 25 µ g/mL TSPN decreased neuronal apoptosis (P<0.05), and immunofluorescence showed that 25 µ g/mL TSPN restored the dendritic morphology of damaged neurons (P<0.05). Moreover, 12.5 µ g/mL TSPN downregulated the expression of Beclin-1, Cleaved-caspase 3 and LC3B-II/LC3B-I, and upregulated the levels of phosphorylated (p)-Akt and p-mTOR (P<0.01 or P<0.05). In the MCAO model, 50 µ g/mL TSPN improved defective neurological behavior and reduced infarct volume (P<0.05). Moreover, the expression of Beclin-1 and LC3B in cerebral ischemic penumbra was downregulated after 50 µ g/mL TSPN treatment, whereas the p-mTOR level was upregulated (P<0.05 or P<0.01).

Conclusion

TSPN promoted neuronal survival and protected dendrite integrity after OGD/R and had a potential therapeutic effect by alleviating neurological deficits and reversing neuronal loss. TSPN promoted p-mTOR and inhibited Beclin-1 to alleviate ischemic damage, which may be the mechanism that underlies the neuroprotective activity of TSPN.

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Acknowledgement

The authors would like to thank the Key Laboratory for providing necessary facilities to ensure that the research was completed. We thank CHEN Ming and LIU Guan-ping for providing and testing the drug. We thank Dr. ZHANG Shi-qing for valuable suggestions on manuscript preparation.

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Authors and Affiliations

  1. JNU-HKUST Joint Laboratory for Neuroscience and Innovative Drug Research, Jinan University, Guangzhou, 510632, China

    Yu-wei Pan, Gen-yun Tang & Lei Shi

  2. Department of TCM Preventive Medicine, Tianhe District Hospital of Traditional Chinese Medicine, Guangzhou, 510632, China

    Yu-wei Pan

  3. Institute of Traditional Chinese Medicine and Natural Products, Guangdong Province Key Laboratory of Pharmacodynamic Constituents of TCM and New Drugs Research, College of Pharmacy, Jinan University, Guangzhou, 510632, China

    Dong-ping Wu, Hua-feng Liang, Chun-lin Fan, Wen-cai Ye & Man-mei Li

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  1. Yu-wei Pan
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Contributions

Li MM, Ye WC and Shi L conceived and designed the study; Wu DP and Liang HF performed the in vivo experiments and collected the data; Pan YW and Tang GY performed the in vitro experiments and collected the data; Fan CL prepared the sample and performed HPLC analysis; Wu DP, Pan YW, Liang HF, Li MM and Shi L wrote the manuscript; Li MM and Shi L revised the work for intellectual content and context; and all the authors read and approved the manuscript.

Corresponding author

Correspondence to Man-mei Li.

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Conflict of Interest

All authors report no conflicts of interest, and no competing financial interests exist.

Supported by the National Key R&D Program of China (No. 2017YFC1703800), the Key-Area Research and Development Program of Guangdong Province (Nos. 2019B030335001 and 2020B1111110004), the Local Innovative and Research Teams Project of the Guangdong Pearl River Talents Program (No. 2017BT01Y036)

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Pan, Yw., Wu, Dp., Liang, Hf. et al. Total Saponins of Panax notoginseng Activate Akt/mTOR Pathway and Exhibit Neuroprotection in vitro and in vivo against Ischemic Damage. Chin. J. Integr. Med. 28, 410–418 (2022). https://doi.org/10.1007/s11655-021-3454-y

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  • DOI: https://doi.org/10.1007/s11655-021-3454-y

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