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Roll of AMPK Reglution in Protein and Cholesterol Level in the Sepsis Rat

Author: XuMi
Tutor: FangQiang
School: Zhejiang University
Course: Internal Medicine
Keywords: AMP-activated protein kinase (AMPK) 3-hydroxy-3-methyl-glutaryl-CoA reductase (HMGR) cholesterol Sepsis mTOR eEF2 Protein regulation
CLC: R459.7
Type: Master's thesis
Year: 2011
Downloads: 62
Quote: 1
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Abstract


Objective:To investigate the relationship of cholesterol level and possible mechanism AMPK pathway in endotoxic rats.Methods:12 wistar rat,were randomly divided into control and LPS groups. Endotoxemia was induced by a slow intravenous injection of 5mg/kg lippolysaccharide via tail vein of rats. Cholesterol level were measured by automatic biochemical analyzer. The protein expression of AMPK and p-AMPK in liver tissue were determined by western blot analysis. The expression of Hydroxymethyl-coenzyme A reductase(HMGR) was assessed by spectrophotometry.Results:Compared to the control group, the level of serum cholesterol in LPS group were markedly elevated (1.50±0.07 in LPS group vs 1.11±0.14 in control group, P＜0.05); In LPS group, the marked increases of p-AMPK protein expression were shown (1.7350±0.46889 in LPS group vs 0.9713±0.23911 in control group, P＜0.05) while LPS failed to increase AMPK protein expression levels of liver tissue; The activity of HMGR was induced to elevate by LPS (0.00026050±0.000047346 in LPS group vs 0.00123114±0.000623238 in control group, P＜0.05);Conclusion:The level of cholesterol in endotoxemia rat induced by LPS was elevated after 24h, meanwhile the expression of p-AMPK protein was activated and the activity of HMGR was depressed. Objective:To study the possible ways of AMPK to regulating protein in sepsis rat.Methods:24 wistar rats were randomly divided into four groups:6h control group,6h LPS group,24h control group and 24h LPS group. N=6, intravenous injection of lipopolysaccharide(LPS 5mg/kg) in LPS group. And injection saline in control group. Specimens from liver and heart tissue samples were detected by western blot. Assay adenosine monophosphate-activated protein kinase (p-AMPK/AMPK), phosphorylated mammalian target of rapamycin (p-mTOR/mTOR), phosphorylation of elongation factor 2 (p-eEF2/eEF2) protein expression.Results:After LPS injection 6 hour, the level of serum total protein, albumin and globulin in LPS group were slightly increased Compared with control group, but not significantly, expression of p-eEF2 both in heart and liver of LPS group decreased compared with control, total eEF2 was no significant difference in both group. expression of p-mTOR in heart was no significant difference and increased in liver, of LPS group compared with control, Total mTOR was no significant difference in both group. expression of p-AMPK both in liver and heart in LPS group has some increased compare with control, but no significant difference in total AMPK.After LPS injection 24 hour, the level of serum total protein and albumin were decreased compare with control, but globulin has no difference.In LPS group the expression of p-eEF2 in both heart and liver wassignificantly higher than the control group, total eEF2 was no significant difference in both of them. expression of p-mTOR in the liver was significantly increased compared with control group. Total mTOR was no significant difference between them.expression of p-AMPK in both has significantly increased compared with control group, but total AMPK was no significant difference.Conclusion:Protein synthesis reduced in LPS-induced sepsis rats after injection LPS 24h, AMPK inhibit protein synthesis by down the activity of eEF2 and mTOR

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