Water extract of medicinal ink (WEMI) attenuates lipopolysaccharide-induced NO production of Raw264.7 cells via downregulating JAK2/STAT3-mediated iNOS expression.

ETHNOPHARMACOLOGICAL RELEVANCE: Medicinal ink is used as a traditional topical medicine for treating inflammatory diseases via detoxification, relieving pain, hemostasis, and reducing swelling. However, the effect of medicinal ink on the inhibition of inflammatory responses and the underlying molecular mechanism remain unclear. AIM OF THE STUDY: The present study aimed to investigate the anti-inflammatory function of water extract of medical ink (WEMI) and elucidate its active mechanisms. MATERIALS AND METHODS: Cell viability was assessed using crystal violet staining assay. Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected by ELISA. Nitric oxide (NO) production was measured by Griess assay. The activation of inflammatory signaling molecules stimulated by lipopolysaccharide (LPS) was evaluated by assessing levels of inducible nitric oxide synthase (iNOS), phosphorylated Janus kinase 2 (JAK2) and signal transducer and activator of transcription 3 (STAT3) using Western blot assay. RESULTS: Water extract of medical ink (WEMI) did not present cytotoxic effect on murine macrophage Raw264.7 cells. High dosage of WEMI slightly rescued LPS-suppressed cell viability of Raw264.7 cells. WEMI did not induce NO production or IL-6 secretion, though WEMI significantly induced secretion of TNF-α on Raw264.7 cells not stimulated with LPS. On the other hand, LPS effectively stimulated inflammation on Raw264.7 cells; however, WEMI dramatically reduced LPS-induced NO production. WEMI alleviated LPS-stimulated IL-6 secretion but did not affect the content of TNF-α. In addition, WEMI effectively reduced expression of iNOS by abolishing LPS-mediated phosphorylation of JAK2 and STAT3 but not TLR4-mediated NF-κB and MAPK molecules. CONCLUSIONS: Our findings suggest that WEMI targets of the JAK2/STAT3-mediated iNOS expression play a key role in alleviating LPS-induced inflammatory responses in RAW264.7 macrophages. Therefore, medicinal ink may be a potential topical agent for treating fasciitis or synovitis via regulating the immune system.

Phytoextraction of uranium from contaminated soil by Macleaya cordata before and after application of EDDS and CA.

This is the first report on using Macleaya cordata for phytoextraction of uranium from the uranium contaminated soil in the greenhouse. Macleaya M. cordata was found to increase uranium concentration in the soil solution by increasing the dissolved organic carbon (DOC). The amendment experiments with citric acid (CA) and [S,S]-ethylenediamine disuccinic acid (EDDS) at the rates of 1.0, 2.5, 5.0, and 10.0 mmol kg(-1) dry weight (DW) soil showed that EDDS was more efficient to increase uranium concentration in the shoot than CA when they were applied at the same rate. The applications of 5.0 mmol kg(-1) EDDS and 10.0 mmol kg(-1) CA were most appropriate for increasing uranium concentrations in the shoot of M. cordata. CA was more efficient to increase the solubility of uranium at the same application rates except for 2.5 mmol kg(-1) application rate. There was a linear correlation between the uranium concentration in the shoot and the average uranium concentration of one planted pot during 14 days in soil solution after the application of different rates of EDDS and CA, respectively (r(2) = 0.972, P < 0.01; r (2) = 0.948, P < 0.01), indicating that uranium uptake was dependent on the soluble uranium concentration. The Fe-U-DOC and Mn-U-DOC complexes were probably formed after the application of CA. Soil solution pH and Fe, Mn, Ca, and DOC concentrations in soil solution were found to be changed by the chelates.

Hepatoprotective effects of aqueous extract from Lingzhi or Reishi medicinal mushroom Ganoderma lucidum (higher basidiomycetes) on α-amanitin-induced liver injury in mice.

The Lingzhi or Reishi mushroom Ganoderma lucidum is a well-known traditional medicinal mushroom that has been shown to have obvious hepatoprotective effects. The aim of this study was to evaluate the hepatoprotective effects of G. lucidum aqueous extracts (GLEs) on liver injury induced by α-amanitin (α-AMA) in mice and to analyze the possible hepatoprotective mechanisms related to radical scavenging activity. Mice were treated with α-AMA prepared from Amanita exitialis and then administrated with GLE after the α-AMA injection. The hepatoprotective activity of the GLE was compared with the reference drug silibinin (SIL). α-AMA induced a significant elevation in serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities and provoked a significant reduction of superoxide dismutase (SOD) and catalase (CAT) activities and a significant increment of malondialdehyde (MDA) content in liver homogenate. Treatment with GLE or SIL significantly decreased serum ALT and AST levels, significantly increased SOD and CAT activities, and decreased MDA content in liver compared with the α-AMA control group. The histopathological examination of liver sections was consistent with that of biochemical parameters. The results demonstrated that GLE induces hepatoprotective effects on acute liver injury induced by α-AMA; these protective effects may be related in part to the antioxidant properties of GLE.

Yin yang-1 regulates the characterized murine focal adhesion-associated protein promoter.

The focal adhesion-associated protein (FAAP), product of the murine D10Wsu52e gene, is involved in modulating cell adhesion dynamics. The ubiquitously expressed protein belongs to the highly conserved UPF0027 family, the newly identified RNA >p ligase family. To understand the mechanisms underlying FAAP expression and regulation, we first mapped its major transcription start site at the nucleotide 79 bp upstream of the ATG codon. The murine FAAP 2.1 kb 5'-flanking region was cloned, analyzed, and aligned with the corresponding 1.7 kb region of its human homolog HSPC117. Despite the differences in activity, cell in vitro transfection and testis in vivo electroporation identified a 0.2 kb efficient promoter region lacking a functional TATA-box. Gel shift assays confirmed the specific interaction between Yin Yang-1 (YY1) and the potential element in the proximal region of the FAAP promoter. Site mutation, truncation, RNAi, and overexpression analyses suggested that YY1 is an important regulator of the FAAP promoter.