| Part 1 Effects of fibrillar Aβ(1-40) on the growth and the Insulin signal transduction pathway related protein of cultured primitive rat basal forebrain cholinergic neuronsObjective1.To investigate the affection of fibrillar Aβ(1-40) to morphology and cellular vigor of cultured primitive rat basal forebrain cholinergic neurons in vitro;2.To investigate the affection of fibrillar Aβ(1-40) to the insulin signal transduction pathway of cultured primitive rat basal forebrain cholinergic neurons in vitro.Methods1.Primitive rat basal forebrain neurons were cultured and evaluated with immuno-fluorescence;2.The cultured neurons were exposed to fibrillar Aβ(1-40) with different ending concentrations(0.1μmol/L,1.0μmol/L,2.0μmol/L,5.0μmol/L & 10μmol/L) and different durations.Then morphologic and cellular vigor changes were observed by fluorescent microscope and MTT assay respectively;3.Investigate the expression of insulin signal transduction pathway related proteins of cultured neurons after the neurons were intervened by fibrillar Aβ(1-40).Results1.After being exposed to fibrillar Aβ(1-40) for 48h,the neurons exposed to low ending concentration(0.1 & 1.0μmol/L) of Aβ(1-40) showed no significant changes in morphology and cellular vigor;while the neurons exposed to higher ending concentration(2.0,5.0& 10.0μmol/L) of Aβ(1-40) showed significant changes in morphology and cellular vigor: average process quantity of the neurons,average process length and the length of the longest process of neurons were decreased significantly,and the results of MTT assay showed that average OD values were significantly decreased.2.The expression of InsR,IRS-1,Akt/PKB,CREB and Bcl-2 in the cultured neurons showed no significant changes after being exposed to Aβ(1-40) with the two lower concentrations(0.1 & 1.0μmol/L);While after being exposed to the three higher concentrations(2.0,5.0&10.0μmol/L) of Aβ(1-40),the expression of InsR,IRS-1,Akt/PKB and Bcl-2 were significantly decreased and the expression of CREB showed no significant change.When the neurons were cultured with 5μmol/L Aβ(1-40) for different duration(24h, 48h,72h),the above indexes were decreased by time dependency except the expression of CREB.ConclusionsAβ(1-40) have a concentration-dependent and time-dependent harmful effect on the activity and insulin signaling pathway of cultured primitive rat basal fore-brain cholinergic neurons, which suggest that the harmful effect to insulin signal transduction pathway induced by fibrillar Aβ(1-40) may contribute to the apoptosis and loss of cholinergic neurons in Alzheimer's disease.Part 2 Study of urinary AD7c-NTP levels in patients with Alzheimer's diseaseObjectiveTo investigate the value of urinary AD7c-NTP assay in the clinical diagnosis of AD.MethodsWith the method of Enzyme-linked immunosorbent assay,to measure urinay AD7c-NTP levels in 245 elderly Chinese people,who were divided into three group including patients with AD(n=65),patients with vascular dementia(VaD,n=60) and normal mental state elderly people(n=120),.Statistical analysis was performed using SPSS program 13.0.Results1.The urinay AD7c-NTP assay of three groups were(2.50±0.45) ng/ml,(1.19±0.68)ng/ml and(1.21±0.74)ng/ml.As compared to normal mental state control and VaD group,there was a significant increase of urinary AD7c-NTP levels in AD group.No significant difference was found between VaD group and normal mental state control; 2.Within the AD group,AD7c-NTP level is positively correlated with the severity of the disease.ConclusionsThe measurement of urinary AD7c-NTP level is of important and practical value in the clinical diagnosis,especially early diagnosis,and Condition assessment of AD.
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