Matrix metalloproteinase mmp 2

Cells were harvested and lysated with radio-Immunoprecipitation (RIPA) buffer (Sigma).
Lysates were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and
then transferred to nitrocellulose membranes (Millipore). Blots were incubated with
primary antibodies and secondary antibodies. Antibodies used in this study included Bcl-2,
caspase-9, matrix metalloproteinase (MMP) 2, and MMP9, from Santa Cruz Biotechnology
(Dallas, Texas); P-38, extracellular signal-regulated kinase (ERK), the c-Jun N-terminal
kinases (JNK), and vascular endothelial growth factor (VEGF) from Abcam (Cambridge,
Massachusetts); and caspase-3 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from
Cell Signaling Technologies. The blots were developed by an enhanced chemiluminescence
(ECL) detection kit (Kaiji Bio Co, Nanjing, China) and visualized under LAS 3000
Luminoimage Analyzer. Three independent experiments were performed, and the representative
results were shown.

The SGC-7901 human gastric cancer cell line was obtained from the Shanghai Institute for Biological Sciences of the Chinese Academy of Sciences (Shanghai, China). The specific inhibitor of ROS, N-acetyl cysteine (NAC), was purchased from Beyotime Institute of Biotechnology (Nantong, China). The inhibitor of CLIC1, indanyloxyacetic (IAA)-94 was purchased from Sigma-Aldrich (St. Louis, MO, USA). The chemical inhibitor of p38-MAPK (SB203580) was purchased from Merck Millipore (Darmstadt, Germany). Antibodies targeting p38 MAPK, phosphorylated (p)-p38 MAPK, matrix metalloproteinase (MMP)-2 and MMP-9 were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA).

Aminophylline was obtained from Shandong Xinhua Pharmaceutical Co., Ltd. (Zibo, People’s Republic of China). Klebsiella pneumoniae (strain ID: 46114) was purchased from the National Center for Medical Culture Collection (Beijing, People’s Republic of China). Antibodies against interleukin (IL)-6, IL-10, tumor necrosis factor (TNF)-α, soluble TNF-α receptor 2, collagen I, collagen III, collagen IV, endothelin (ET)-1, transforming growth factor (TGF)-β, vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), matrix metalloproteinase (MMP)-2, MMP-9, and tissue inhibitor of MMP (TIMP)-1 were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). The RNeasy kit was obtained from Qiagen (Valencia, CA, USA). Mayer’s hematoxylin and 1% eosin alcohol solution were purchased from MUTO Pure Chemicals (Tokyo, Japan). In all, 42 Sprague Dawley rats (21 male and 21 female; 200±20 g) were purchased from the Experimental Animal Center of Henan Province (Zhengzhou, People’s Republic of China). The animals were housed in cages with free access to food and tap water under standard conditions of humidity (50%±10%), temperature (25°C±2°C), and light (12 hours light/12 hours dark cycle). All animals were handled with humane care throughout the experiment.