ABSTRACT
AIM:To investigate the regulatory effect of NADPH oxidase-4 (NOX-4) on PI3K signaling path-way in transforming growth factor-β1 (TGF-β1)-induced collagen type Ⅰ (collagen Ⅰ) synthesis from lung cancer cells and the mechanisms. METHODS:Human lung cancer A549 cells were cultured in vitro and stimulated with TGF-β1. The ex-pression of NOX family and collagen family at mRNA and protein levels as well as the PI3K class Ⅰ catalytic subunits and the activation of PI3K signaling pathway was measured. A549 cells were pre-treated with NOX-4 inhibitor diphenyleneiodo-nium (DPI), and the expression of collagen Ⅰ at mRNA level as well as the PI3K class Ⅰ catalytic subunits and the activa-tion of PI3K signaling pathway was measured upon TGF-β1 stimulation. RESULTS:TGF-β1 stimulated the expression of NOX-4 and collagen Ⅰ at mRNA and protein levels as well as the expression of PIK3CD and the activation of PI3K signaling pathway at a dose- and time-dependent manner. NOX-4 inhibitor DPI partly reversed TGF-β1-induced collagen Ⅰ expres-sion. Inhibition of NOX-4 down-regulated the degree of TGF-β1-stimulated activation of PI3K signaling pathway without effect on the expression of PIK3CD. CONCLUSION:NOX-4 participates in TGF-β1-induced collagen Ⅰ synthesis from lung cancer cells via regulating the activation of PI3K signaling pathway. TGF-β1/NOX-4/PI3K signaling pathway axis acts as a regulatory role in collagen Ⅰ synthesis from lung cancer cells.
ABSTRACT
Cysteine-rich angiogenic inducer 61(Cyr61/CCN1)is an extracellular matrix-associated signaling protein consisting of 381 amino-acid residues ,which has the regulatory function for a multitude of cellular responses.The pleiotropic effects of CCN 1 on the initiation and resolution of inflammation as well as oncogenesis and development of tumor were reported.According to the numerous data from experimental and clinical studies ,this article provides an overview on CCN1 and summarizes the latest understanding of the role of CCN 1 in pulmonary diseases.
ABSTRACT
<p><b>OBJECTIVE</b>To observe the effect of pretreatment with puerarin on activation of LPS -induced RAW264. 7 cells and secretory cytokines, and discuss its anti-inflammatory mechanism.</p><p><b>METHOD</b>Well-grown RAW264. 7 cells in the exponential phase were collected and randomly divided them into the blank control group, the LPS group and the puerarin pretreatment + LPS group. The cellular toxic effect of puerarin on RAW264. 7 cells was examined by CCK-8 assay, cell morphology was detected by Giemsa stain method, the changes in TNF-alpha and MIP-2 were tested by ELISA, and the expression of NF-kappaB p65 mRNA were determined by qRT-PCR.</p><p><b>RESULTS</b>When puerarin was cultured with 1 mg x L(-1) LPS at a concentration of lower than 400 micromol x L(-1), it had not showed the cellular toxic effect (P < 0.05). Compared with the control group, the LPS group could significantly change the morphology of RAW264. 7 cells (increase in cell body, irregular shape, with a large number of pseudopodia extending). After intervention, the puerarin 100 micromol x L(-1) group could significantly inhibit LPS-induced cell morphological changes, while the puerarin 200 micromol x L(-1) and 400 micromol x L(-1) puerarin groups showed more notable inhibitory effects. However, there was no obvious difference between the two groups. The pretreatment with puerarin could inhibit the expression of TNF-alpha and MIP-2 in cell supernatant and NF-kappaB p65 mRNA in cells (P < 0.05). With increase in the puerarin concentration, its inhibitory effect gradually grew (P < 0.05), but did not reach the level of the blank control group.</p><p><b>CONCLUSION</b>As a safe and effective natural anti-inflammatory drug, puerarin can significantly reduce the expression of inflammatory cytokines (TNF-alpha, MIP-2). Its mechanism may be related to the reduction of NF-kappaB p65 mRNA expression.</p>
Subject(s)
Animals , Mice , Cell Line , Isoflavones , Pharmacology , Lipopolysaccharides , Allergy and Immunology , Macrophage Activation , Macrophages , Allergy and Immunology , NF-kappa B , Genetics , Allergy and Immunology , Plant Extracts , Pharmacology , Sincalide , Genetics , Allergy and Immunology , Transcription Factor RelA , Genetics , Allergy and Immunology , Tumor Necrosis Factor-alpha , Genetics , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of mito chondrial K(ATP) channels (mitoK(ATP)) inhibitor 5-hydroxydecanoate(5-HD) on chronic hypoxic pulmonary artery hypertension (CHPAH) rats and its underlying mechanisms.</p><p><b>METHODS</b>Forty-eight male SD rats were equally divided into 4 groups randomly (n=12): normal group, hypoxia group, hypoxia + 5-HD group, hypoxia + Diazoxide group. Except the first group, the other three groups were put into hypoxic [O2 (10.0% +/- 0.3%] and nonrmobaric chamber for four weeks to establish chronic hypoxic model and received different interference. When the interference completed, right heart catheter was used to detect the mean pulmonary arterial pressure (mPAP) of each rat and PKC-alpha mRNA expression in pulmonary arteries was detected by reverse transcription-polymerase chain reaction (RT-PCR) and protein expression by Western blot.</p><p><b>RESULTS</b>(mPAP was much higher in hypoxia group than that in normal group (P < 0.01) while in hypoxia + 5-HD group and hypoxia + diazoxide were decreased significantly compared to hypoxia group (P < 0.01). (2) The protein and mRNA levels of PKC-alpha in the hypoxic group were higher than those in normal group (P < 0.05).</p><p><b>CONCLUSION</b>5-HD plays a protective role on CHPAH. The mechanism of its effect may be attributed to inhibiting MitoK(ATP).</p>