ABSTRACT
Alzheimer's disease(AD) patients in China have been surging, and the resultant medical burden and care demand have a huge impact on the development of individuals, families, and the society. The active component compound of Epimedii Folium, Astragali Radix, and Puerariae Lobatae Radix(YHG) can regulate the expression of iron metabolism-related proteins to inhibit brain iron overload and relieve hypofunction of central nervous system in AD patients. Hepcidin is an important target regulating iron metabolism. This study investigated the effect of YHG on the expression of a disintegrin and metalloprotease-17(ADAM17), a key enzyme in the hydrolysis of β amyloid precursor protein(APP) in HT22 cells, by mediating hepcidin. To be specific, HT22 cells were cultured in vitro, followed by liposome-mediated siRNA transfection to silence the expression of hepcidin. Real-time PCR and Western blot were performed to examine the silencing result and the effect of YHG on hepcidin in AD cell model. HT22 cells were randomized into 7 groups: control group, Aβ25-35 induction(Aβ) group, hepcidin-siRNA(siRNA) group, Aβ25-35 + hepcidin-siRNA(Aβ + siRNA) group, Aβ25-35+YHG(Aβ+YHG) group, hepcidin-siRNA+YHG(siRNA+YHG) group, Aβ25-35+hepcidin-siRNA+YHG(Aβ+siRNA+YHG) group. The expression of ADAM17 mRNA in cells was detected by real-time PCR, and the expression of ADAM17 protein by immunofluorescence and Western blot. Immunofluorescence showed that the ADAM17 protein expression was lower in the Aβ group, siRNA group, and Aβ+siRNA group than in the control group(P<0.05) and the expression was lower in the Aβ+siRNA group(P<0.05) and higher in the Aβ+YHG group(P<0.05) than in the Aβ group. Moreover, the ADAM17 protein expression was lower in the Aβ+siRNA group(P<0.05) and higher in the siRNA+YHG group(P< 0.05) than in the siRNA group. The expression was higher in the Aβ+siRNA+YHG group than in the Aβ+siRNA group(P<0.05). The results of Western blot and real-time PCR were consistent with those of immunofluorescence. The experiment showed that YHG induced hepcidin to up-regulate the expression of ADAM17 in AD cell model and promote the activation of non-starch metabolic pathways, which might be the internal mechanism of YHG in preventing and treating AD.
Subject(s)
Humans , ADAM17 Protein , Alzheimer Disease/genetics , Amyloid beta-Peptides , Drugs, Chinese Herbal/pharmacology , Hepcidins/genetics , PuerariaABSTRACT
Aim To explore the effect of a combination of Astragalus, Epimedium and Pueraria on the expression of hepcidin in hippocampal CA3 region of APPswe/PSldE9 double transgenic AD model mice. Methods Thirty 10-month-old APPswe/PSldE9 double transgenic model mice were randomly divided into three groups; the compound group, the model group and the deferox (D F X) group, and ten 10-month-old C57BL/6J mice were as normal control group. Subsequently, the brain tissue of the mice was taken out, and the expression of hepcidin in the hippocampal CA3 region of each group of mice was detected by immunofluorescence combined with Real-time PCR and Western blot. Results Compared withnormal group, the mRNA expression of hepcidin was down-regulated inmodel group. Afterintragastric administrationof active components of Epimedium, Astragalus and Radix Puerariae and DFX, hepcidin expression levels increased significantly compared with those in APPswe/PSldE9 double transgenic AD model mice hippocampal CA3 area (P 0. 05). Conclusions The effective components of Astragalus, Epimedium and Radix Puerariae can up-regulate the expression of hepcidin, It is suggested that the effective components of Epimedium, Astragalus, and Radix Puerariae have important theoretical significance in the clinical treatment of Alzheimer's disease with iron overload.