Protection and mechanism of neoeriocitrin on Aβ 25-35 injured PC12 cells / 国际中医中药杂志

ABSTRACT

Objective:To study the protective effect of neoeriocitrin on PC12 cell injury induced by amyloid β protein fragment 25-35 (Aβ 25-35), and explore its role in preventing and treating AD. Methods:The MTT method was used to detect the effect of Neoeriocitrin on the proliferation of normal PC12 cells and Aβ 25-35 damaged PC12 cells, and the intervention concentration of neoeriocitrin solution was screened. The PC12 cells were divided into the normal control group, model group, estradiol group and neoeriocitrin group according to the random number table method. The normal control group and model group were cultured with DMEM for 24 h; DMEM and 1×10 -3 μmol/L estradiol solution were added to the estradiol group; DMEM and 6×10 4 μmol/L neoeriocitrin solution were added to the neoeriocitrin group. After 2 hours of culture, except for the normal control group, the remaining groups were added with 20 μmol/L Aβ 25-35 stock solution, and the culture was continued for 22 h. AnnexinV-FITC/PI double labeling was used to detect apoptosis rate, Western blot method was used to detect estrogen receptor β (estrogen receptor β, ERβ) and p-P38/P38 protein expression. The content of acetylcholine (ACh) was detected, and the activity of Choline acetyl transferase (ChAT) and Acetylcholin esterase (AChE) in the supernatant of PC12 cells were detected. Results:Compared with the model group, the cell apoptosis rate [(8.080 ± 0.578)% vs. (18.500 ± 0.870)%] significantly decreased ( P<0.01), the expression of ERβ protein (0.348 ± 0.042 vs. 0.273 ± 0.006) significantly increased ( P<0.01), p-P38/P38 protein expression (0.372 ± 0.058 vs. 0.571 ± 0.063) significantly decreased ( P<0.01), the content of ACh [(14.319 ± 1.039) μg/mg vs. (9.157 ± 1.605) μg/mg], ChAT activity [(0.715 ± 0.053) U/mg vs. (0.280 ± 0.093) U/mg] significantly increased ( P<0.01), AChE activity [(2.607 ± 2.048) U/mg vs. (6.038 ± 1.867) U/mg] significantly reduced ( P<0.01). Conclusions:Neoeriocitrin can promote the proliferation of PC12 cells damaged by Aβ25-35, inhibit cell apoptosis, and has a certain cytoprotective effect. Its mechanism may be related to the regulation of ERβ expression and inhibition of P38 protein phosphorylation, thereby improving the function of the cholinergic system related.