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 μg/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 μg/mL TSPN decreased neuronal apoptosis(P＜0.05),and immunofluorescence showed that 25 μg/mL TSPN restored the dendritic morphology of damaged neurons(P＜0.05).Moreover,12.5 μg/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 μg/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 p.g/mL TSPN treatment,whereas the p-mTOR level was upregulated(P＜0.05 or P＜0.01).Conclusions: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.

PAN Yu-wei;WU Dong-ping;LIANG Hua-feng;TANG Gen-yun;FAN Chun-lin;SHI Lei;YE Wen-cai;LI Man-mei

JNU-HKUST Joint Laboratory for Neuroscience and Innovative Drug Research,Jinan University,Guangzhou,510632,China;Department of TCM Preventive Medicine,Tianhe District Hospital of Traditional Chinese Medicine,Guangzhou,510632,China;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

中国结合医学杂志（英文版）

[1]PAN Yu-wei;WU Dong-ping;LIANG Hua-feng;TANG Gen-yun;FAN Chun-lin;SHI Lei;YE Wen-cai;LI Man-mei-.Total Saponins of Panax notoginseng Activate Akt/mTOR Pathway and Exhibit Neuroprotection in vitro and in vivo against Ischemic Damage)[J].中国结合医学杂志（英文版）,2022(05):410-418

Saponins,TSPN,stainings

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