Network Pharmacology Analysis and Molecular Characterization of the Herbal Medicine Formulation Qi-Fu-Yin for the Inhibition of the Neuroinflammatory Biomarker iNOS in Microglial BV-2 Cells: Implication for the Treatment of Alzheimer’s Disease

Figure 3

Bioactivity-guided fractionation of the herbal formulation QFY for potential active compounds. (a) Schematic illustration of the bioactivity-guided fractionation procedure. The fractions were prepared by sequential water extraction, ethanolic precipitation, and RP-HPLC separation on a C18 column. The fractions were sequentially analyzed by Western blot for the effects on LPS-induced expression of iNOS and COX-2 in BV-2 cells. (b) Bioassays of the QFY water extract and the fractions from ethanolic precipitation. BV-2 cells were treated with the indicated QFY extract with or without simultaneous stimulation with LPS for 24 h. The expression of iNOS and COX-2 was detected by Western blot using specific antibodies. Representative blots were shown. The blots () were quantified by a densitometric method using the ImageJ software. The results were expressed as . (LPS vs. untreated control); (treatment+LPS vs. LPS). (c) HPLC separation of the 90% QFY fraction. The compounds were separated into nine fractions by RP-HPLC on a C18 column. (d) Bioassays of the QFY-derived HPLC fractions. BV-2 cells were treated with the indicated concentration of different HPLC fractions or 90% QFY together with LPS stimulation. The cellular proteins were analyzed by Western blot, and the blots were quantified as previously described. (LPS vs. untreated control); , , and (treatment+LPS vs. LPS).

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