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  • The Role of Oxymatrine in Amelioration of Acute Lung Injury Subjected to Myocardial I/R by Inhibiting Endoplasmic Reticulum Stress in Diabetic Rats.

The Role of Oxymatrine in Amelioration of Acute Lung Injury Subjected to Myocardial I/R by Inhibiting Endoplasmic Reticulum Stress in Diabetic Rats.

Evidence-based complementary and alternative medicine : eCAM (2020-12-10)
Yongpan Huang, Xian Long, Xinliang Li, Saihua Li, Jianbin He
ABSTRACT

Oxymatrine (OMT) is the primary pharmacological component of Sophora flavescens Aiton., which has been shown to possess potent antifibrotic, antioxidant, and anti-inflammatory activities. The aim of the present study was to clarify the protective mechanism of OMT on acute lung injury (ALI) subjected to myocardial ischemia/reperfusion (I/R). A myocardial I/R-induced ALI model was achieved in diabetic rats by occluding the left anterior descending coronary artery for 1 h, followed by reperfusion for 1 h. The levels of inflammatory factors (tumor necrosis factor-α, interleukin- (IL-) 6, and IL-17) in bronchoalveolar lavage fluid were assessed using commercially available kits. The index of myocardial injury, including the detection of cardiac troponin I (cTnI), cardiac troponin T (cTnT), lactate dehydrogenase (LDH), and creatine kinase-MB (CK-MB), was also determined using commercially available kits. Hematoxylin and eosin staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling were used to identify histological changes. The expression levels of endoplasmic reticulum chaperone BiP (GRP78), DNA damage-inducible transcript 3 protein (CHOP), eukaryotic translation initiation factor 2-alpha kinase 3 (PERK), inositol dependent enzyme 1α (IRE1α), ATF6, caspase-3, -9, and-12, Bcl-2, and Bax were determined by Western blotting. The mRNA expression levels of GRP78 and CHOP were detected by reverse transcription-quantitative PCR. Myocardial I/R increased the levels of cTnI, cTnT, LDH, and CK-MB in diabetic rats. Damaged and irregularly arranged myocardial cells were also observed, as well as more serious ALI with higher lung injury scores and WET/DRY ratios and lower PaO2. Moreover, the expression of key proteins of endoplasmic reticulum stress (ERS) was increased by I/R injury, including phosphorylated- (p-) PERK, p-IRE1ɑ, and ATF6, as well as decreased levels of apoptosis. These effects were all significantly reversed by OMT treatment. OMT protects against ALI subjected to myocardial I/R by inhibiting ERS in diabetic rats.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-phospho-PEK/PERK (pThr981) antibody produced in rabbit, affinity isolated antibody
Roche
Red Blood Cell Lysis Buffer, solution, Roche, pkg of 100 mL, sufficient for 50-500 reactions
Sigma-Aldrich
Rat IL-6 ELISA Kit, for serum, plasma and cell culture supernatants
Sigma-Aldrich
Caspase 3 Assay Kit, Colorimetric
Sigma-Aldrich
Monoclonal Anti-LDH (H-Subunit) antibody produced in mouse, clone HH-17, ascites fluid
Sigma-Aldrich
Anti-PERK (N-terminal) antibody produced in rabbit, IgG fraction of antiserum, buffered aqueous solution
Sigma-Aldrich
Anti-CHOP, N-Terminal antibody produced in rabbit, affinity isolated antibody
Sigma-Aldrich
Anti-GRP78/BiP (GL-19) antibody produced in rabbit, IgG fraction of antiserum, buffered aqueous solution
Sigma-Aldrich
Anti-Caspase 9 antibody produced in rabbit, affinity isolated antibody
Roche
In Situ Cell Death Detection Kit, Fluorescein, sufficient for ≤50 tests, suitable for detection
Anti-Cardiac Troponin I (cTnI), in vitro, clone 1611, clone 1611, from mouse
Sigma-Aldrich
Anti-Protein Kinase Cα antibody produced in rabbit, whole antiserum
Sigma-Aldrich
Tumor Necrosis Factor-α from rat, TNF-α, recombinant, expressed in E. coli, powder, suitable for cell culture
Sigma-Aldrich
Monoclonal Anti-Caspase 12 antibody produced in rat, clone 14F7, purified immunoglobulin, buffered aqueous solution
Sigma-Aldrich
Anti-Caspase 3 antibody produced in rabbit, IgG fraction of antiserum, buffered aqueous solution
Sigma-Aldrich
Monoclonal Anti-Bcl-2 antibody produced in mouse, clone Bcl-2-100, ascites fluid
Sigma-Aldrich
Anti-ATF6 (ab1) antibody produced in rabbit, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Creatine Kinase Activity Assay Kit, sufficient for 100 tests (Ultraviolet)