Study On Selective Estrogen Receptor Modulators(SERMs) For Neuro-protecting A? Cells Model Of Alzheimer's Disease

Alzheimer's disease(AD)is the most common neurodegenerative disease in the old.With the development of aging population structure in modern society,AD has a very important and prominent position in the disease and death,and has become the fourth class of death after heart disease,cancer,stroke.Currently,there are many kinds of AD drugs are appearing as therapeutic options in clinic,such as nucleoside analogues and interferon,such as antagonist of glutamate receptor,the target drugs of amyloid ?-protein(A?),estrogen replacement therapy etc.However,existing drugs are relied on the prevention and mitigation of progression of disease,such as the toxicity and side effect.According to new research,the disease mainly in postmenopausal women,due to the changes of estrogen in postmenopausal women,estrogen in the body can play a role by binding to the receptor,and it has the function of improving memory and improving cognitive ability.However,estrogen replacement therapy has many disadvantages,long drug administration,significant risk of endometrial carcinoma and mammary carcinoma.Selective estrogen receptor modulators(SERMs)is similar to the effect of estrogen,it show different effects by combining with estrogen receptor(ER)in different tissues to excite the central nervous system,and become the replacement of estrogen.Based on the above theory,In this study,SERMs was used to establish the model of neuroprotection and detected of neuroprotective effects of drugs.PC12 cells is necessary to induce into nerve cells.When the A?1-42 damage model was established,testing the neuroprotective effects of SERMs on the representative drug tamoxifen.1 Induced differentiation of PC12 into neurons.We use the methods of cell morphology,Real-time PCR,Western blot to test neural markers and choose optimal screening condition.It was found that the gene level of nerve markers increased by 1.2-2 times,icluding the UCH-L1,NFP and MAP2.In each treatment group of NGF,icreased expression of MAP2 protein will change wth the increase of NGF dose.The optimal concentration of NGF was 50?g/L.2 Establishment of nerve cell injury model.Cell viability was detected by CCK8 assay,the morphological changes of cells apoptosis were observed by microscope.Changes in the potential energy of mitochondrial membrane potential by JC-1.It was found that A?1-42 can cause cell morphology,cell apoptosis and cell mitochondrial membrane potential decreased significantly.The optimal damage concentration of A?1-42 was 20?mol/L.3 Detection of the protective effect of tamoxifen on the expression of SERMs.Cell viability was detected by CCK8 assay,Hoechst 33258 staining was used to observe the cell morphology,changes in the potential energy of mitochondrial membrane potential by JC-1.The changes of Bax and Bcl-2 expression of apoptosis protein were detected by western blot.The results of the experiment show that estrogen and TAM have neuroprotective effect on the model.TAM can prevent the decrease of mitochondrial membrane potential,and has a protective effect on mitochondria.Conclusion: In this study,we have completed the establishment of cell model of differentiation and A? damage.We find that TAM has neuroprotective function.