[Effect of active component compound of Epimedii Folium,Astragali Radix,and Puerariae Lobatae Radix on expression of ADAM17 in HT22 cells by mediating hepcidin].

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 ß 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, Aß25-35 induction(Aß) group, hepcidin-siRNA(siRNA) group, Aß25-35 + hepcidin-siRNA(Aß + siRNA) group, Aß25-35+YHG(Aß+YHG) group, hepcidin-siRNA+YHG(siRNA+YHG) group, Aß25-35+hepcidin-siRNA+YHG(Aß+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 Aß group, siRNA group, and Aß+siRNA group than in the control group(P<0.05) and the expression was lower in the Aß+siRNA group(P<0.05) and higher in the Aß+YHG group(P<0.05) than in the Aß group. Moreover, the ADAM17 protein expression was lower in the Aß+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 Aß+siRNA+YHG group than in the Aß+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.

[Effect of lactobacilli on Th1/Th2 cells balance in primary lymphocytes].

AIM: To analyse the effect of 7 Lactobacillus species on proliferation and cytokine production of primary lymphocytes, and explored the effect on the Th1/Th2 cells balance. METHODS: The mouse spleen cells were stimulated with active/heat-killed Lactobacillus strains at different species and concentrations. After be cultured with bacteria 60 hours, the proliferation of splenocytes were tested by MTT colorimetry, and IL-12, IFN-γ, IL-4, IL-10 and TGF-ß were detected by ELISA. RESULTS: (1) Both alive and heat-killed Lactobacillus species could accelerate the proliferation of spleen cells in a dose-dependent manner (P<0.05), the immuneological activity of heat-killed L.fermentum and L.acidophilu were not inferior to the live strains, especially when treated at higher dose(10(7);CFU/mL, bacteria: cell ratio of 10:1). Furthermore, these two heat-killed bacterial strains suppressed IL-4, IL-10 and TGF-ß production by stimulating IL-12 and IFN-γ production, and their IFN-γ/IL-4 value which represent the Th1/Th2 balance were higher than ConA control group significantly(P<0.05). CONCLUSION: The results indicate that Lactobacillus strains may differ in their respective ability to improve Th1/Th2 cells balance toward Th1 dominance via increasing the secretion ratio of IFN-γ/IL-4 (Th1/Th2) by splenocytes.