Tamarix hohenackeri Bunge exerts anti-inflammatory effects on lipopolysaccharide-activated microglia in vitro

Abstract

Background: Tamarix species are well known as the main host plants of Herba Cistanches, a valuable Traditional Chinese Medicine. They are also traditional medicinal plants themselves and are used to treat spleen problems, leucoderma and ocular conditions.

Purpose: The aim of the present study was to investigate the anti-inflammatory effect of Tamarix hohenackeri Bunge.

Methods: In the present study, BV-2 microglial cells were used and stimulated with lipopolysaccharide (LPS). Cell viability was tested using the MTT assay. The release of nitric oxide (NO) was determined using the Griess assay. The mRNA level of inducible nitric oxide synthase (iNOS), tumor necrosis factor α (TNF-α), interleukin (IL)-1β and IL-6 were investigated by quantitative real-time PCR (qRT-PCR). The protein levels of phosphorylated of IκBα, ERK and MEK, as well as the cytoplasmic and nuclear NF-κB p65 were tested by Western blot analysis. The translocation of the NF-κB p65 subunit from the cytosol to the nucleus was investigated by immunofluorescence staining.

Results: Ethyl acetate (EtOAc) extract of Tamarix hohenackeri Bunge significantly inhibited the release of NO. Phytochemical research was performed to produce 13 main constituents. Among them, compounds 6, 7, 10 and 13 were identified to be the effective components with anti-inflammatory activity. These compounds significantly inhibited the production of NO by LPS-activated BV-2 microglial cells. qRT-PCR showed that compounds 6 and 7 significantly suppressed the LPS-induced transcription of genes encoding pro-inflammatory mediators, including iNOS, TNF-α, IL-1β and IL-6. Western blot analysis showed that compound 7 inhibited the LPS-induced phosphorylation of IκBα and antagonized the LPS-induced reduction of cytoplasmic NF-κB p65 and the increase of nuclear NF-κB p65. Immunofluorescence staining showed that nuclear translocation of NF-κB p65 was suppressed by compound 7. Western blot analysis showed that compound 7 inhibited the LPS-induced phosphorylation of ERK and MEK.

Conclusion: The present study revealed, for the first time, the effective anti-inflammatory agents from T. Hohenackeri. Compound 7 exerted potent anti-inflammatory effects and its underlying mechanism may be associated with its capacity to inhibit NF-κB signaling pathway and the MEK/ERK activation in activated microglia. The compound may be potential candidate therapeutic agent for neurodegenerative diseases.

Keywords: Anti-inflammatory; LPS; Microglia; Neurodegenerative diseases; Tamarix hohenackeri Bunge.