PMID: 16524497Mar 10, 2006Paper

Effect of dexamethasone on degradation of cultured myotube protein and its mechanism

Zhongguo wei zhong bing ji jiu yi xue = Chinese critical care medicine = Zhongguo weizhongbing jijiuyixue
Chuan-an ShenZhi-yong Sheng

Abstract

To study the effect of dexamethasone on degradation of long-lived protein in myotubes and to elucidate its possible mechanism. After isolating skeletal muscles of rat's hind limb under sterile condition, the myoblasts were proliferated in tissue-block culture. After being passaged and purified, the myoblasts fused into myotubes. The protein in myotubes was radiolabelled with L-[3,5-(3)H]-tyrosine, and then the myotubes were divided into two groups. In group A, the myotubes were cultured in Dubbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS) and 2 mmol/L tyrosine without dexamethasone (control group), with dexamethasone 10 nmol/L (group A1), dexamethasone 100 nmol/L (group A2), and dexamethasone 1,000 nmol/L (group A3), respectively. The amounts of L-[3,5-(3)H]-tyrosine in culture medium and cells were determined and the degradation rates of long-lived protein were calculated at 12, 24, 36, 48 hours of culture. In group B, the myotubes were cultured in DMEM containing 10% FBS and 2 mmol/L tyrosine with 50 micromol/L proteasome inhibitor MG132 (group B1), or 50 micromol/L MG132 and 100 nmol/L dexamethasone (group B2). Proteolytic rates of protein were calculated with the same way as group A at 24 hou...Continue Reading

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