Aβ25-35 Led HT22 Cells In The Endoplasmic Reticulum Stress And Protective Effect Of TSG

Objective: Alzheimer’s disease is a common type of dementia in the elderly。It is mainly for degenerative diseases of the nervous system of cognitive dysfunction and behavioral characteristics of abnormal. The main pathological feature of AD is a large number of senile plaques and neurofibrillary tangles in the brain tissue. The main component of senile plaques is Aβ, so neurotoxicity of Aβ is the key factor for the formation and development of AD. Researchs show that changes of expression of endoplasmic reticulum stress pathway,s protein, will lead to cell apoptosis and cause disease. In this experiment, the main purpose is to investigate Aβ25-35 induced HT22 cells model of AD cells. To explore Aβ25-35 led HT22 cells in the endoplasmic reticulum stress and protective effect of TSG.Method: HT22 was divided into undifferentiated group and differentiated group.The undifferentiated group was cultured for 48 hours with complete culture medium.The differentiated group was cultured for 24 hours with complete culture medium,following the differentiation medium(Neurobasal medium and N2 additive) was added for 24 hours.The expression of IRE1,PERK and ATF6 in the two groups were detected by Western Blot. After determining the optimal AD cell model, Differentiated HT22 was divided into control group、Aβ group and Aβ +TSG group. The differentiated group was cultured for 24 hours with complete culture medium,following the differentiation medium(Neurobasal medium and N2 additive) was added for 24 hours. Different concentrations of Aβ 25-35 role in HT22 cells 24 hours, MTT determinate in cell viability. Immune cell fluorescence check the endoplasmic reticulum stress three pathways of protein expression and Western blotting were used to detect the ATF6,PERK and IRE1 protein expression changes.Result:1. The expression of ATF6, PERK and IRE1 in the undifferentiated group was significantly higher than that in the differentiation group. Aβ25-35 on undifferentiated group cell is injured, but the injury is not in accordance with the increase of drug concentration increased.Aβ25-35 can induce differentiation of HT22 cells that were damaged, and the damage is in accordance with the increase of drug concentration increased. Aβ25-35(20μmol/L 、 40μmol/L)effects on HT22 cell differentiation for 24 hours, which the cell survival rate were 66% and60%.In this case, the establishment of the cell model is the best AD model.2. Differentiated HT22 was divided into control group、Aβ group and Aβ +TSG group.The differentiated group was cultured for 24 hours with complete culture medium,following the differentiation medium(Neurobasal medium and N2 additive) was added for 24 hours. Different concentrations of Aβ25-35 role in HT22 cells 24 hours, MTT determinate in cell viability.Immune cell fluorescence check the endoplasmic reticulum stress three pathways of protein expression and Western blotting were used to detect the ATF6,PERK and IRE1 protein expression changes.Conclusion:1. Aβ25-35(20μmol/L) induced differentiated HT22 cells to establish the model of AD cells.2. Aβ25-35 can led HT22 cells in the endoplasmic reticulum stress and TSG has a protective effect on it.