Protective effect of total flavonoids extract of Rhodiola rosea L.on Aβ25-35 induced PC12 cell damage and screening of active parts
Objective To determine the protective effect of total flavonoids extract of Rhodiola rosea L.(TFE)on PC12 cell damage induced by amyloid β-protein 25-35(Aβ25-35)and screen its active parts.Methods PC12 cell damage was induced by 20 μmol·L-1 Aβ25-35,and TFE was isolated by AB-8 macroporous adsorption resin chromatography.Water and 20%,40%,60%,80%and 95%ethanol elution(TFE-0%,TFE-20%,TFE-40%,TFE-60%,TFE-80%and TFE-95%)were obtained.The extract yield of ethanol elution with different volume fractions(0.3 weight)was calculated.The total flavonoids content of ethanol elution with different volume fractions(0.7 weight)was detected by UV spectrophotometry,and the ethanol elution with different volume fractions was evaluated comprehensively.The effects of different volume fractions of ethanol elution on the viability of injured PC12 cells were detected by MTT method.The morphology of cells was observed by inverted microscope,the activity of lactate dehydrogenase(LDH)was detected by 2,4-dinitrophenylhydrazine colorimetric method,and the content of superoxide dismutase(SOD)by xanthine oxidase method.The levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),cyclooxygenase-2(COX-2)and interleukin-1β(IL-1β)in the supernatant were detected by enzyme-linked immunosorbent assay.Results The comprehensive scores of total flavonoids concentration and extract yield of TFE-40%eluates were higher than those of other ethanol eluents with different volume fractions.Compared with the blank group,the intercellular space of the model group was larger,the number of cells decreased,the cell fragments appeared,the synapse decreased significantly,the survival rate and the level of SOD decreased significantly(P＜0.05),and the level of LDH increased significantly(P＜0.05).Compared with the model group,the cell morphology of TFE-40%,TFE-60%and Donepezil hydrochloride groups improved to some extent,the number of synapses increased,with more complete cell morphology.The levels of IL-6,TNF-α,COX-2,and IL-1β in the supernatant of TFE-40%,TFE-60%,TFE-80%and TFE-95%groups were significantly decreased(P＜0.05).Conclusion The TFE-40%eluent of Rhodiola rosea L.has a good protective effect on the injury of PC12 cells induced by Aβ25-35,which is the best active part.The mechanism may be related to the neurotoxicity and inhibition of cellular inflammation of antagonism of Aβ25-35.
Rhodiola rosea L.total flavonoidPC12protective effectactive part
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