Pang M.,Shanxi Medical University |
Wang H.,Shanxi Medical University |
Bai J.-Z.,University of Auckland |
Cao D.,Shanxi Medical University |
And 7 more authors.
Experimental Biology and Medicine | Year: 2015
Clara cell protein (CC16) is a well-known anti-inflammatory protein secreted by the epithelial Clara cells of the airways. It is involved in the development of airway inflammatory diseases such as chronic obstructive pulmonary disease and asthma. Previous studies suggest that CC16 gene transfer suppresses expression of interleukin (IL)-8 in bronchial epithelial cells. However, its role in the function of these cells during inflammation is not well understood. In this study, we evaluated the effect of CC16 on the expression of matrix metalloproteinase (MMP)-9 in lipopolysaccharide (LPS)-stimulated rat tracheal epithelial cells and its underlying molecular mechanisms. We generated recombinant rat CC16 protein (rCC16) which was bioactive in inhibiting the activity of phospholipase A2. rCC16 inhibited LPS-induced MMP-9 expression at both mRNA and protein levels in a concentration-dependent (0–2 µg/mL) manner, as demonstrated by real time RT-PCR, ELISA, and zymography assays. Gene transcription and DNA binding studies demonstrated that rCC16 suppressed LPS-induced NF-κB activation and its binding of gene promoters as identified by luciferase reporter and gel mobility shift assays, respectively. Western blotting and immunofluorescence staining analyses further revealed that rCC16 concentration dependently inhibited the effects of LPS on nuclear increase and cytosol reduction of NF-κB, on the phosphorylation and reduction of NF-κB inhibitory IκBα, and on p38 MAPK-dependent NF-κB activation by phosphorylation at Ser276 of its p65 subunit. These data indicate that inhibition of LPS-mediated NF-κB activation by rCC16 involves both translocation- and phosphorylation-dependent signaling pathways. When the tracheal epithelial cells were pretreated with chlorpromazine, an inhibitor of clathrin-mediated endocytosis, cellular uptake of rCC16 and its inhibition of LPS-induced NF-κB nuclear translocation and also MMP-9 production were significantly abolished. Taken together, our data suggest that clathrin-mediated uptake of rCC16 suppresses LPS-mediated inflammatory MMP-9 production through inactivation of NF-κB and p38 MAPK pathways in tracheal epithelial cells. © 2015, by the Society for Experimental Biology and Medicine.
Liao H.,Shanxi Provincial Peoples Hospital |
Banbury L.,Southern Cross University of Australia |
Wang X.,Shanxi Da Yi Hospital |
Lu X.,Shanxi Provincial Peoples Hospital |
And 2 more authors.
Journal of Traditional Chinese Medicine | Year: 2014
OBJECTIVE: To study the effects of extracts from Honghua (Flos Carthami) on lipopolysaccharide induced nitric oxide (NO) production in RAW 264.7 cells and the influence of the extracts on yeast α-glucosidase activity. The total flavonoid content of the extracts was also determined. METHODS: Cytotoxicity of the extracts to RAW 264.7 cells was evaluated by the ATPlite™ method. Inhibitory effects of the extracts on NO production were evaluated by Griess assay. Curcumin was used as a positive control. Screening of extracts for potential α-glucosidase inhibitors was done by a fluorometric assay. The assay was based on the hydrolysis of 4-methylumbelliferyl-a-D-glucopyranoside to form the fluorescent product, 4-methylumbelliferone. Acarbose was used as a positive control. The total flavonoid content was tested using kaempferol as the standard. RESULTS: There were significant inhibitory effects on NO production when the extracts were 25-100 μg/ mL (P<0.05) and curcumin was 2-4 μg/mL (P< 0.001). The extracts showed an inhibitory effect on α-glucosidase activity at the concentrations of 15.6-125 μg/mL with a half maximal (50%) inhibitory concentration (IC50) of (32.8 ± 5.7) μg/mL, compared with the IC50 of acarbose at (1.8±0.4) μg/mL. There was a significant difference between the two IC50 values (P<0.001). The total content of flavonoids per gram of dried herb was 1.14 mg. CONCLUSION: Honghua (Flos Carthami) showed inhibitory effects on NO production in activated RAW 264.7 macrophage cells and an inhibitory effect on yeast α-glucosidase. There might be a relationship between these pharmacological effects and its flavonoid content. © 2014 JTCM. All rights reserved.
Chen L.,Shanxi Da Yi Hospital |
Tong H.,Shanxi Da Yi Hospital |
Yang Z.,Shanxi Da Yi Hospital
Journal of Bionanoscience | Year: 2013
Fourth ventricular dystopia tumor is an extremely rare tumor. We are reporting a case of fourth ventricular dystopia tumor in a girl, where total extirpation of the lesion was performed, pertinent literature, regarding on the lesions located in the posterior fossa midline, has been reviewed. The differential diagnosis of Fourth ventricular lesions is discussed. Copyright © 2013 American Scientific Publishers.