title: Attenuation of cellular toxicity by calpain inhibitor induced by bacterial endotoxin: a mechanistic study using muscle precursor cells as a model system.
abstract: This investigation was under taken to explore probable mechanisms and signal pathways involved in cytotoxicity induced by bacterial endotoxin lipopolysaccharide (LPS). Herein, we selected muscle precursor C2C12 myoblasts as representative cells to test effect of calpain inhibitor 3-(4-iodophenyl)-2-mercapto-(Z)-2-propenoic acid (PD150606) on LPS induced inflammation and apoptosis. In order to rule out the toxicity of endotoxin, mouse myoblasts were exposed to various concentrations of LPS and viability of cells and morphology were assessed using CCK-8 assay and simple microscopy respectively. Apoptotic cell death was examined by fluorescence microscope at regular time intervals. Additionally, LPS induced apoptosis in C2C12 cells were determined by mRNA expression of µ-calpain, caspase-3 and tumor necrosis factor alpha (TNF-α) and were quantified by qRT-PCR. Our results point out that LPS stimulation produced dose dependent toxicity in muscle precursor cells. Pre-treatment with a calpain inhibitor can significantly attenuate LPS-induced inflammation/apoptosis. Results of present research determined that mRNA expression of aforesaid genes was amplified (p<0.05) in LPS stimulated C2C12 cells, whereas a noticeable drop off in mRNA expression of these genes was observed when pre-exposed with calpain inhibitor PD150606. Our study has outlined the current understanding regarding the connection between µ-calpain and caspase-3 in skeletal muscle wasting and as a result provides suitable choice for designing promising chemotherapeutic system for muscle illness and atrophy.
authors: Department of Animal Science and Biotechnology, Chonbuk National University, Chonju, 561-756, Republic of Korea.
authors: Ke
authors: K
authors: Shang
authors: Junfeng
authors: J
authors: Zhang
authors: Touseef
authors: T
authors: Amna
authors: Jieun
authors: J
authors: Yang
authors: Xiangchao
authors: X
authors: Cheng
authors: Chunjie
authors: C
authors: Zhang
authors: Inho
authors: I
authors: Hwang
mesh_terms: D000179:Acrylates / Q000494:pharmacology*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D003603:Cytotoxins / Q000633:toxicity*; D006023:Glycoproteins / Q000494:pharmacology*; D007249:Inflammation; D008070:Lipopolysaccharides / Q000633:toxicity*; D051379:Mice; D008954:Models, Biological; D032446:Myoblasts / Q000187:drug effects* / Q000502:physiology; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D000179:Acrylates; D003603:Cytotoxins; D006023:Glycoproteins; D008070:Lipopolysaccharides; C100442:PD 150606; C039304:calpain inhibitors