Objective To investigate the effects of nicotinic acetylcholine receptors(nAChRs)α7 on the proliferation and apoptosis of human neuroblastoma cells(SH-SY5Y),and to clarify that α7nAChR can exert its biological function by positively regulating the expression of dopamine(DA)neurons,so as to treat Parkinson's disease.Methods In this experiment,SH-SY5Y cell line was selected for cell culture,resuscita-tion,passage,production of Parkinson's cell model and cell transfection.The experimental subjects were di-vided into control group,model group,α 7nAChR overexpression group,and α 7nAChR overexpression no-load group.CCK8 was performed to detect cell activity.Flow cytometry was used to detect apoptosis.The ex-pression of dopamine(DA)was detected by ELISA.The protein expressions of α7nAChR,p-CAMK Ⅱ,p-ERK and p-Ras were detected by Western Blot Analysis.The expressions of TH,α7nAChR and αSYN were detected by immunofluorescence.Results The protein expression level of α7nAChR in Model group was sig-nificantly lower than that in Control group(P＜0.05).The protein expression level in α7nAChR-OE group was significantly higher than that in Model group(P＜0.05).The results of CCK8 showed that the cell prolif-eration activity in Model group was significantly decreased compared with that in control group(P＜0.05).The cell proliferation activity in α7nAChR-OE group was significantly higher than that in Model group,P＜0.05.Flow cytometry showed that the apoptosis rate of cells in Model group was significantly higher than that in Control group(P＜0.05).The apoptosis rate of α7nAChR-OE group was significantly decreased compared with that in Model group(P＜0.05).The αSYN in Model group was significantly increased compared with that in Control group(P＜0.05).The αSYN in α7nAChR-OE group was significantly decreased compared with that in Model group(P＜0.05).The TH,α7nAChR and DA in Model group were significantly decreased compared with those in Control group(P＜0.05).The TH,α7nAChR and DA in α7Nachr-OE group were significantly increased compared with those in Model group(P＜0.05).The protein phosphorylation levels of KRAS,CAMK Ⅱ and ERK in Model group were significantly increased compared with those in Control group(P＜0.05).The protein phosphorylation levels of KRAS,CAMK Ⅱ and ERK in α7nAChR-OE group were significantly decreased compared with those in Model group(P＜0.05).Conclusion The high expression ofα7nAChR can promote the proliferation and inhibition of the apoptosis in Parkinson's model cells.Besides,α7nAChR can play a neuroprotective role by inhibiting the activity of Ca2+/CAMK/ERK pathway,increasing the expression of DA neurons,and improving Parkinson's disease.
维普
万方数据