The Effect Of Liuwei Dihuang Wan On Aβ-induced Autophagy In SH-SY5Y Cells And Its Mechanism

Purpose:To study the effect and mechanism of Liuwei Dihuang Pill serum on autophagy of SH-SY5 Y cells induced by Aβ,to explore the partial pathological mechanism of Alzheimer’s disease and the partial mechanism of kidney tonifying and essence filling method,and to provide experimental basis for the prevention and treatment of Alzheimer’s disease.Material and method:30 rats were randomly divided into two groups: control group and Liuwei Dihuang Pill group 1.18g/kg Liu Wei Di Huang Wan powder water solution was given to the stomach twice a day for 5 days.Blood was taken 2 hours after the end of the last gastric perfusion,and serum was separated for use as blank serum and drug containing serum.SH-SY5 Y cells were divided into three groups: control group,model group and drug group.After the cells adhered to the wall,the control group and model group were cultured in complete culture medium containing 10% blank serum,the drug group was cultured in complete culture medium containing 10% drug serum,the model group and drug group were cultured in Aβ 1-42 oligomer with final concentration of 20 μ mol / L for 48 h.CCK-8 method was used to analyze the cell proliferation rate of each group;transmission electron microscope was used to observe the cell morphology and the number of autophagy;immunofluorescence method was used to analyze the Aβ deposition in each group;Western blot method was used to detect the expression of BACE1,Atg7,p62,LC3-Ⅰ,LC3-Ⅱ,Beclin1 protein in each group.Results:1.CCK-8 results showed that compared with the control group,the cell proliferation rate of the model group was significantly lower(p < 0.01);compared with the model group,the cell proliferation rate of the drug group was significantly higher(p < 0.01).2.The results of immunofluorescence showed that compared with the control group,the deposition of Aβ in the model group increased significantly(p < 0.01);compared with the model group,the deposition of Aβ in the drug group decreased significantly(p < 0.01).3.Transmission electron microscopy showed that compared with the control group,the morphology of cells in the model group was seriously damaged and the number of autophagy increased;compared with the model group,the morphological changes of cells in the drug group were significantly reduced and the number of autophagy increased.4.Western Blot results showed that compared with the control group,the protein expression of BACE1,Atg7,Beclin1 and the ratio of LC3-Ⅱ /LC3-Ⅰ in the model group were significantly higher(p < 0.01),and the protein expression of p62 was significantly lower(p <0.01);compared with the model group,the protein expression of Atg7,Beclin1 and the ratio of LC3-Ⅱ/LC3-Ⅰ in the drug group were significantly higher(p < 0.01),and the protein expression of BACE1 and p62 were significantly reduced(p < 0.01).Conclusion:1.Liuwei Dihuang Pill serum has obvious protective effect on SH-SY5 Y cells induced by Aβ.2.Liuwei Dihuang Pill serum can up regulate the autophagy level of SH-SY5 Y cells induced by Aβ,reduce the deposition of Aβ in cells,and reduce cell damage.