Organic Extract of Justicia pectoralis Jacq. Leaf Inhibits Interferon-γ Secretion and Has Bacteriostatic Activity against Acinetobacter baumannii and Klebsiella pneumoniae

Abstract

Justicia pectoralis Jacq. (Acanthaceae) leaves currently found in the Brazilian north-east are widely used to treat diabetes, menstrual pains, asthma, and other disorders. This work aimed to identify the phytochemical characterization and biological activities of J. pectoralis leaf extracts. The plant material was ground and the crude extracts were obtained with water or acetone: water (7:3 v/v), yielding aqueous (JPA), and organic (JPO) extracts. Phytochemical characterization was performed by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). Cytotoxicity was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT) assay and trypan blue (TB) exclusion assay in peripheral blood mononuclear cells (PBMCs), BALB/c splenocytes, and neoplastic cells (TOLEDO, K562, DU-145, and PANC-1) at 1, 10, and 100 μg/mL. Antibacterial activity was evaluated using the microdilution test to obtain the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Cytokines, IFN-γ, and IL-17A from culture supernatants of BALB/c mice splenocytes were measured by sandwich ELISA. In the TLC analysis, both JPA and JPO extracts presented coumarin and flavonoids. In addition, HPLC was able to identify coumarin, apigenin, and ellagic acid in both extracts. JPO IC₅₀ was 57.59 ± 1.03 μg/mL (MTT) and 69.44 ± 8.08 μg/mL (TB) in TOLEDO. MIC value of JPO against Acinetobacter baumannii and Klebsiella pneumoniae was 500 μg/mL. JPO (100 μg/mL) significantly inhibited IFN-γ levels (p=0.03). J. pectoralis is a potential candidate to be further investigated as an IFN-γ inhibitory agent and against Acinetobacter baumannii and Klebsiella pneumoniae.

Figure 1

Chromatographic profile of JPA and JPO and UV-sprectra of peaks related (coumarin, apigenin, and ellagic acid). Detection at 280 nm. Justicia pectoralis aqueous extract (JPA); Justicia pectoralis organic (JPO) extracts.

Figure 2

Justicia pectoralis cytotoxic evaluation against neoplastic cell lines TOLEDO, K562, PANC-1, and DU-145 at 1, 10, and 100 μg/mL of JPA and JPO. MTT test are shown at (a), (c), (e), (g), and trypan blue are present at (b), (d), (f), and (h).

Figure 3

Cytokines titration in culture supernatant of Balb/c splenocytes treated with aqueous (JPA) and organic extract (JPO) of Justicia pectoralis at 10 and 100 μg/mL. (a) IFN-γ levels; (b) IL-17A levels (UC = untreated control, CONA = concanavalin A (5 μg/mL), and MP = methylprednisolone 100 μM). ∗ p < 0.05. ● represents outliers.