Levels of sAPP�� and sAPP��-sw were reduced by 58% Ixazomib clinical (p<0.01) and 70% (p<0.001), respectively, by treatment with 12.5 ��M berberine, and by 30% (p<0.05) and 45% (p<0.01) by 6.25 ��M berberine. Berberine treatment at a concentration of 12.5 ��M reduced the level of Fl-APP, pAPPThr668 and CTFs by 54% (p<0.01), 42% (p<0.05) and 67% (p<0.01), respectively. Since RS and HLJDT increased all APP metabolic products, we ascertained whether baicalein alone can induce the same APP increasing effect as did RS and HLJDT. As hypothesized, baicalein significantly increased soluble APPs, Fl-APP, pAPPThr668 and CTFs in a dose-dependent manner (Figure 5B). The levels of maximal increase of Fl-APP, pAPPThr668 and CTFs by baicalein were 1.68 (p<0.05), 1.58 (p<0.01) and 2.36 (p<0.
05) fold of basal release, respectively, at a concentration of 12.5 ��M. The release of sAPPs was accelerated by treatment with baicalein in a dose-dependent manner, reaching maximal secretion of 2.65 (p<0.01) and 1.70 (p<0.05) fold of basal release for sAPP�� and sAPP��-sw, respectively, at a concentration of 12.5 ��M. These data suggest that berberine is one of the alkaloids responsible for the APP-decreasing effect of HLJDT-M, and baiclein is one of the flavonoids responsible for the APP-increasing effect of HLJDT. Figure 5 Modulation of APP processing by berberine and baicalein in N2a-SwedAPP cells. Modulation of Intracellular A�� Levels by HLJDT, HLJDT-M and its Components in N2a-SwedAPP Cells Since HLJDT and its components modulated intracellular levels of APP and CTFs in N2a-SwedAPP, we investigated the effect of HLJDT and its components on the level of intracellular A��, which is a key factor in AD progression [29].
We treated N2a-SwedAPP cells with different non-toxic concentrations of HLJDT, HLJDT-M and its components and then analyzed intracellular A�� levels by ELISA (Figure 6). There was a dose-dependent reduction in both A��1�C40 and A��1�C42 by both RC and CP treatments. Intracellular A��1�C40 markedly dropped 41% and 29% due to treatment by 25 ��g/mL RC and CP, respectively (Figure 6A). There was a 32% reduction in intracellular A��1�C42 in both RC and CP treated cells at the same high concentration. In contrast, RS treatment increased intracellular A��s in a dose-dependent manner, reaching maximal accumulation of 1.4- and 1.6- fold of basal levels of A��1�C40 and A��1�C42, respectively, at a concentration of 1.
56 ��g/mL (Figure AV-951 6B). Figure 6 Regulation of the levels of intracellular A�� by HLJDT, HLJDT-M and its components in cultured N2a-SwedAPP cells. When comparing the effects of HLJDT and HLJDT-M on the level of intracellular A��s, as expected, HLJDT increased both A��1�C40 and A��1�C42 in a dose-dependent manner, reaching maximal accumulation of 1.4- and 1.6- fold of basal level, respectively, at a concentration of 12 ��g/mL (Figure 6C).