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 4

Identification of the active ingredients for the downregulation of iNOS expression. (a) Bioassays of the QFY extract and individual herbal extracts. QFY and individual herbal ingredients were extracted with water and precipitated with 90% ethanol. BV-2 cells were treated with 90% ethanol solutions of herbal extracts for 24 h with LPS stimulation. The expression of specific markers was detected by Western blot, and the blots were quantified as previously described. (LPS vs. untreated control); (treatment+LPS vs. LPS). (b) HPLC separation of the 90% RS fraction. The 90% RS solution was separated into nine fractions by RP-HPLC on a C18 column using the same gradient and elution time as described for the 90% QFY fraction. (c) Bioassays of the 90% RS-derived HPLC fractions. BV-2 cells were treated with the indicated concentration of HPLC fractions or 90% RS for 24 h with LPS stimulation. The expression of specific markers was detected by Western blot, and the blots were quantified as previously described. , (LPS vs. untreated control); (treatment+LPS vs. LPS).

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