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The Effect of Amylin on the Gene Expression of Human Hepatocyte Glucose-6-phosphatase

Author: LiShiYing
Tutor: ZhuTieHong
School: Tianjin Medical University
Course: Internal Medicine
Keywords: Glucose - 6 - phosphatase Hepatocytes Amylin Insulin resistance Semi - quantitative RT-PCR
CLC: R587.1
Type: Master's thesis
Year: 2008
Downloads: 70
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Abstract


Objective : glucose - 6 - phosphatase ( glucose - 6 -phosphatase , G- 6 - Pase ) is an important enzyme in the process of sugar metabolism , because it is the rate-limiting enzyme of the last reaction step of gluconeogenesis and glycogenolysis , changes in gene expression levels can affect the the endogenous sugar output . In fact, type 2 diabetes, high blood sugar with hepatic insulin resistance due to excessive hepatic glucose output related . Amylin as important active hormone of pancreatic beta cells , involved in blood sugar regulation . Studies have shown that abnormal secretion involved in insulin resistance . The purpose of this experiment is to study the amylin gene expression of the enzyme , to further explore the correlation of amylin and hepatic glucose output and insulin resistance . Method : the introduction of human liver cell line L - 02 , according to the literature were cultured in vitro . 2 different concentrations of glucose, insulin and different concentrations of amylin medium cultured hepatocytes , and then extract the total cellular RNA , the application of semi - quantitative reverse transcription polymerase chain reaction ( SQ - RT - PCR ) method to detect G-6 - Pase expression levels . 1 by the difference between the microscope hepatocyte growth . 2 human liver cells cultured in high glucose ( 15 mM ) , insulin (100IU / L) solution after 24 hours of incubation , G-6-Pase mRNA abundance was higher than the control group ( Diet low insulin group ) , with statistical significance . 3 different concentrations of amylin with high glucose and insulin co-culture after 2 hours , 0.1 mu m of 1μM amylin Group G - 6 - Pase mRNA did not change significantly , of the 10μM amylin causes the G - 6 - Pase mRNA higher abundance of glucose and insulin were significantly higher , with statistical significance . Conclusion: a human liver L-02 cells were cultured in vitro . 2 in high glucose and insulin medium for 24 hours , compared with the control group ( low sugar low insulin group ) , increased the expression levels of G - 6 - Pase , suggesting that a high concentration of glucose-lowering insulin on G - 6 - Pase inhibition of mRNA ; different concentrations of amylin , with high glucose and insulin group compared to 0.1 mu m , 1μM amylin group G the - - PasemRNA level no significant change ; the 10μM amylin make G- the 6 - PasemRNA expression levels were significantly increased ( P < 0.01 ) , high concentrations of amylin , high glucose conditions to further reduce the insulin inhibition of G-6-PasemRNA . Prompt diabetes abnormal secretion of amylin interferes with hepatic glucose output , involved in insulin resistance .

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CLC: > Medicine, health > Internal Medicine > Endocrine diseases and metabolic diseases > Islet disease > Diabetes
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