Anti-oxidizing effect of the dichloromethane and hexane fractions from Orostachys japonicus in LPS-stimulated RAW 264.7 cells via upregulation of Nrf2 expression and activation of MAPK signaling pathway

Acid-sensing ion channels (ASICs) are proton-gated cation channels widely expressed in the nervous system. Proton sensing by ASICs has been known to mediate pain, mechanosensation, taste transduction, learning and memory, and fear. In this study, we investigated the differential subcellular localization of ASIC2a and ASIC3 in heterologous expression systems. While ASIC2a targeted the cell surface itself, ASIC3 was mostly accumulated in the ER with partial expression in the plasma membrane. However, when ASIC3 was co-expressed with ASIC2a, its surface expression was markedly increased. By using bimolecular fluorescence complementation (BiFC) assay, we confirmed the heteromeric association between ASIC2a and ASIC3 subunits. In addition, we observed that the ASIC2a-dependent surface trafficking of ASIC3 remarkably enhanced the sustained component of the currents. Our study demonstrates that ASIC2a can increase the membrane conductance sensitivity to protons by facilitating the surface expression of ASIC3 through herteromeric assembly. [BMB Reports 2016; 49(10): 542-547]

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“…Western blotting was performed as previously described (32). Briefly, the protein samples were separated by SDS-PAGE using 10% gels.…”

Section: Methodsmentioning

confidence: 99%

ASIC2a-dependent increase of ASIC3 surface expression enhances the sustained component of the currents

et al. 2016

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“…The O. japonicus was separated sequentially with organic solvents. These soluble fractions were examined for the effect of anticancer in various cells such as human gastric, hepatoma, colon, ovarian, and pancreatic cancer (Kim, Nam, Kim, Ryu, & Lee, ; Lee, Lee, Kim, Kim, et al, ; Lee et al, ; Lee, Lee, Kim, Suk, et al, ; Ryu, Lee, Kwon, & Lee, ; Ryu et al, ). Among these fractions, EtOAc and DCM extracts exhibited the highest effect for apoptosis signaling pathways.…”

Section: Resultsmentioning

confidence: 99%

“…Fractionated O. japonicus was supplied from a farm in Miryang (Geobugiwasong Ltd.). The O. japonicus was separated using organic solvents, and the extract method was described in the previous studies (Lee, Lee, Kim, Kim, et al, ; Lee, Kim, & Lee, ; Lee, Lee, Kim, Suk, et al, ; Ryu et al, ). Each fraction removed the solvents by evaporator at 40°C to dryness.…”

Section: Methodsmentioning

confidence: 99%

“…Orostachys japonicus is a polygamous plant in the family Crassulaceae that has been used as a medicinal herb for fever, hemostasis, antidote, inflammation, and cancer (Kim, Choi, Park, Lee, & Jung, ; Kwon & Han, ; Lee, Lee, Kim, Kim, et al, ; Lee, Ryu, et al, ; Lee, Lee, Kim, Suk, et al, ; Ryu, Lee, Lee, & Lee, ; Yoon et al, ). The previous studies have reported that O. japonicus has beneficial ingredients, such as flavonoids, triterpene, and galactic acid (Park, Han, Park, Choi, & Choi, ).…”

Section: Introductionmentioning

confidence: 99%

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Orostachys japonicus extract inhibits the lipopolysaccharide‐induced pro‐inflammatory factors by suppression of transcription factors

Lee

2020

Food Science & Nutrition

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Orostachys japonicus (O. japonicus) was extracted with ethanol (EtOH) and sequentially separated with organic solvents, including n‐hexane (Hex), dichloromethane (DCM), ethyl acetate (EtOAc), n‐butanol (BuOH), and water (H2O). All the fractions were confirmed for anti‐inflammatory activity in an inflammatory condition. The DCM fraction showed the highest anti‐inflammatory ability. Here, we examined the effect of DCM fraction and investigated the intracellular signaling pathways in LPS‐stimulated RAW 264.7 macrophage cells. The DCM fraction significantly inhibited the mRNA levels of pro‐inflammatory mediators and cytokines including iNOS, COX‐2, IL‐1β, IL‐2, IL‐6, and IP‐10 in LPS‐stimulated cells. Also, the treatment of DCM fraction excellently reduced the expression of the proteins of AP‐1 (phospho‐c‐Jun and phospho‐c‐Fos) and phospho‐IRF3 as transcription factors. As a result, it suppressed LPS‐induced inflammatory mediator and cytokines via inhibition of transcription factors. In conclusion, our data demonstrated that DCM fraction has a strong anti‐inflammatory activity that improves the inflammatory state.

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“…The culture supernatants were collected at various time points and stored at −80 °C until use. After the incubation, the cell viability was measured by the MTS cell proliferation assay (One Solution Cell Proliferation Assay, Promega Corporation, WI, USA) according to the manufacturer’s instructions40. Briefly, 20 μl of MTS were added to the culture plates, incubated for 2 h at 37 °C in a 5% CO 2 incubator, and then read with a microplate photometer at a wavelength of 490 nm.…”

Section: Methodsmentioning

confidence: 99%

The role of macrophages in the susceptibility of Fc gamma receptor IIb deficient mice to Cryptococcus neoformans

Surawut

Ondee

Taratummarat

et al. 2017

Sci Rep

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Dysfunctional polymorphisms of FcγRIIb, an inhibitory receptor, are associated with Systemic Lupus Erythaematosus (SLE). Cryptococcosis is an invasive fungal infection in SLE, perhaps due to the de novo immune defect. We investigated cryptococcosis in the FcγRIIb−/− mouse-lupus-model. Mortality, after intravenous C. neoformans-induced cryptococcosis, in young (8-week-old) and older (24-week-old) FcγRIIb−/− mice, was higher than in age-matched wild-types. Severe cryptococcosis in the FcγRIIb−/− mice was demonstrated by high fungal burdens in the internal organs with histological cryptococcoma-like lesions and high levels of TNF-α and IL-6, but not IL-10. Interestingly, FcγRIIb−/− macrophages demonstrated more prominent phagocytosis but did not differ in killing activity in vitro and the striking TNF-α, IL-6 and IL-10 levels, compared to wild-type cells. Indeed, in vivo macrophage depletion with liposomal clodronate attenuated the fungal burdens in FcγRIIb−/− mice, but not wild-type mice. When administered to wild-type mice, FcγRIIb−/− macrophages with phagocytosed Cryptococcus resulted in higher fungal burdens than FcγRIIb+/+ macrophages with phagocytosed Cryptococcus. These results support, at least in part, a model whereby, in FcγRIIb−/− mice, enhanced C. neoformans transmigration occurs through infected macrophages. In summary, prominent phagocytosis, with limited effective killing activity, and high pro-inflammatory cytokine production by FcγRIIb−/− macrophages were correlated with more severe cryptococcosis in FcγRIIb−/− mice.

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Anti-oxidizing effect of the dichloromethane and hexane fractions from Orostachys japonicus in LPS-stimulated RAW 264.7 cells via upregulation of Nrf2 expression and activation of MAPK signaling pathway

Cited by 28 publications

References 24 publications

ASIC2a-dependent increase of ASIC3 surface expression enhances the sustained component of the currents

ASIC2a-dependent increase of ASIC3 surface expression enhances the sustained component of the currents

Orostachys japonicus extract inhibits the lipopolysaccharide‐induced pro‐inflammatory factors by suppression of transcription factors

The role of macrophages in the susceptibility of Fc gamma receptor IIb deficient mice to Cryptococcus neoformans

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