Effect of Jian Nao Zeng Zhi Yin on neuroprotection and apoptosis in AD cell model
Objective To investigate the neuroprotective effect of Jian Nao Zeng Zhi Yin on Alzheimer's disease(AD)cell model and its effect on apoptosis.Methods PC12 cells were treated with 20 μmol/L amyloid-β(Aβ1)-42 for 24 h and divided into six groups:normal cell control group(NC group),AD cell model group(AD group),positive drug control AD group(AD+Donepezil group)and Jian Nao Zeng Zhi Yin(low concentration,medium concentration and high concentration)treatment AD group(AD+5 μg/ml Jian Nao Zeng Zhi Yin group,AD+10 μg/ml Jian Nao Zeng Zhi Yin group,AD+20 μg/ml Jian Nao Zeng Zhi Yin group).Cell viability was detected by CCK-8 assay;the expression levels of apoptotic factor-related proteins Bax and Bcl-2 were detected by Western blotting;and apoptosis was determined by flow cytometry.Results The cellular activity was tested by CCK-8 assay,and the results showed that the cellular activity in AD group was significantly decreased compared with the NC group(P＜0.05);compared with the AD group,the cellular activity in AD+Donepezil group,AD+20 μg/ml Jian Nao Zeng Zhi Yin group was also significantly increased(P＜0.05).The expression levels of apoptotic factor-related proteins were tested by western blotting,and the results showed that the expression level of Bax was significantly up-regulated,Bcl-2 was significantly down-regulated and Bcl-2/Bax was significantly down-regulated in AD group compared with the NC group(P＜0.05);compared with the AD group,Bax in AD+Donepezil group,AD+10 μg/ml Jian Nao Zeng Zhi Yin group and AD+20 μg/ml Jian Nao Zeng Zhi Yin group was significantly down-regulated,while Bcl-2/Bax in AD+Donepezil group and AD+20 μg/ml Jian Nao Zeng Zhi Yin group was significantly up-regulated(P＜0.05).The apoptosis was tested by flow cytometry,and the results showed that the apoptosis rate was(7.870±0.241)%in NC group,(36.530±1.429)%in AD group,(10.690±1.257)%in AD+Donepezil group,(26.930±1.790)%in AD+5 μg/ml Jian Nao Zeng Zhi Yin group,(23.800±1.136)%in AD+10 μg/ml Jian Nao Zeng Zhi Yin group,(10.410±1.903)%in AD+20 μg/ml Jian Nao Zeng Zhi Yin group.Compared with the NC group,the apoptosis rate of AD group was significantly increased(P＜0.05);compared with the AD group,the apoptosis rate of AD+5 μg/ml Jian Nao Zeng Zhi Yin group,AD+10 μg/ml Jian Nao Zeng Zhi Yin group and AD+20 μg/ml Jian Nao Zeng Zhi Yin group was significantly decreased(P＜0.05).Conclusion Jian Nao Zeng Zhi Yin may reduce apoptosis of AD cell and increase cell viability by regulating the expression of Bcl-2/Bax protein,and play a neuroprotective role in AD cell model.High concentration(20 μg/ml)of Jian Nao Zeng Zhi Yin treatment group has a better efficacy,which can provide a theoretical and clinical basis for the further exploration of the traditional Chinese medicine treatment of cognitive disorders.
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