Acanthopanax Senticosus Saponins Attenuates Lead-induced Cytotoxicity And Apoptosis In PC12 Cells

Objective:Acanthopanax Senticosus has the function of anti-fatigue,anti-oxidation,anti-inflammatory,preventing memory decay and enhancing immunity.According to research,Acanthopanax senticosus saponins have pharmacological effects such as central nervous system excitation,anti-aging,lowering blood lipid,lowering blood sugar,anti-inflammatory and analgesic,anti-aging and anti cancer.Nowadays,Acanthopanax Senticosus saponins have been widely used in the development and research of traditional Chinese medicine in treating many diseases and health products.In this paper,the protective effects of Acanthopanax senticosus saponins(ASS)on lead-induced PC 12 cells were studied from two aspects of cell toxicity and cell apoptosis.Methods:Nerve growth factor(NGF)-differentiated PC 12 cells were pretreated 4h by 12.5mg/L,25mg/L,50mg/L ASS,and then treated with 10μmol/L lead in 24h.The protective effect of ASS on the cytotoxicity of lead-induced PC 12 cells cytotoxicity by using Thiazole Blue Colorimetric method(MTT),lactate dehydrogenase(LDH)content determination,Malondialdehyde(MDA)content determination and nitric oxide(NO)content determination method.And the protective effect of ASS on cell apoptosis induced by lead was detected by Annexin V-FITC apoptosis detection,Western-blot assay Caspase-3,GSK-3β and P-GSK-3β(Ser 9)protein level,and Hoechst 33342 immunofluorescence assay.Results:Compared with the blank control group,the MTT metabolic rate,LDH release,MDA,and NO content in the lead model group were significantly increased.Compared with the lead model group,the MTT metabolic rate of the low,middle and high doses of ASS increased,the LDH release and MDA content decreased,and the content of NO in the middle and high doses of ASS decreased significantly.Annexin V-FITC cell apoptosis assay results showed that compared with the lead model group,the apoptosis rate of the middle and high doses of ASS decreased.Western blot analysis showed that compared with the blank control group,the expression levels of Caspase-3(activated),GSK-3β and P-GSK-3β(ser 9)protein were significantly increased in the lead model group,compared with the lead model group.The expression levels of Caspase-3(activating)protein in the low,medium,and high doses of ASS were significantly lower,and the expression of GSK-3β protein was decreased in the low dose group of ASS;the middle and high doses of Acanthopanax senticosus were lower.P-GSK-3β(ser 9)protein expression levels decreased.The results of Hoechst 33342 immunofluorescence assay showed that compared with the blank control group,the number of stained cells and the staining intensity of the lead model group increased significantly;compared with the lead model group,the ASS low,middle,and high dose groups and with the Acanthopanax senticosus With the increase of saponin dosage,the number and brightness of stained cells were significantly reduced.Conclusion:Acanthopanax senticosus saponins can reduce the cytotoxicity of lead-induced PC 12 cells and reduce apoptosis.Acanthopanax senticosus saponins have protective effects on the cytotoxicity and apoptosis of lead-induced PC12 cells.