Authors

Chunli Wu*, huili liu, Yuan liu, Yongxing Yan

Departments

Department of Neurology, The 3rd Hospital of Hangzhou City

Abstract

Objective: To explore the effect of savatiside A (SA) on the cells of Aβ-42-induced Alzheimer’s disease mice and calcium balance regulation-related proteins and its mechanisms.

Methods: SH-SY5Y neurons were cultured, and those at logarithmic phase were divided into control group (without special treat- ment), Aβ-42 group (modeled with Aβ-42 at a concentration of 30 μM), and SA group (added SA at a concentration of 100μM based on Aβ-42 group). Cell relative viability was calculated by MTT colorimetry. The cells were stained following the Gimesa kit operating instructions and cell morphology observed under a microscope. The cells were stained by Fluo-3AM staining, and the intracellular cal- cium concentration was measured by a flow cytometer. The expression levels of calcium balance-related regulatory proteins Calnexin and p-AMPK2 were detected by Western Blotting method.

Results: group (P<0.01). The survival rate of cells was significantly higher in SA group than that in the Aβ-42 group (P<0.01). The SH-SY5Y neurons in the control group had regular morphology with large volume, and the cell nuclear was regular round and cen- tralized, and cells were tightly arranged and even stained. In the Aβ-42 group, the refractive index of SH-SY5Y neurons was decreased significantly, and the cells were crimpled, aggregated and tangled. The morphology of neurons in the SA group recovered significantly and the number of neurons increased. The findings of fluorescence intensity revealed that the mean fluorescence intensity was much higher in the Aβ-42 group than that in the control group (P<0.05). The average fluorescence intensity was much lower in the SA group than that in the Aβ-42 group (P<0.05). The expression of Calnexin and p-AMPK2 protein was much higher in the Aβ-42 group than that in the control group (P<0.01). The expression of Calnexin and p-AMPK2 protein was significantly lower in the SA group than that in the Aβ-42 group (P<0.01).

Conclusion: Savatiside A has a significant protective effect on Alzheimer's disease-induced mouse cells induced by Aβ-42, which may be associated with regulating calcium balance in neurons and decreasing the expression of Calnexin and p-AMPK2.

Keywords

savatiside A, Alzheimer's disease, calcium balance regulation related protein, Aβ-42.

DOI:

10.19193/0393-6384_2019_6_461