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Ameliorative Effects and Mechanism of Rehmannia Glutinosa Oligosaccharides on Insulin Resistance of HepG2 Cell

Author: GuoLiMin
Tutor: JiaZhengPing；ZhangRuXue
School: Lanzhou University
Course: Biochemistry and Molecular Biology
Keywords: Rehmannia glutinosa oligosaccharides HepG2 Insulin Resistance Peroxisome proliferator-activated receptor α Glucose transporter 2
CLC: R285.5
Type: Master's thesis
Year: 2007
Downloads: 398
Quote: 4
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Abstract


Objective: Chinese medicine Rehmannia extract - dihuang, oligosaccharides using HepG2 cells, the use of modern pharmacology and molecular biology research methods to explore research Rehmannia glutinosa oligosaccharides improve the role of hepatic insulin resistance and molecular mechanisms . Methods: MTT (MTT) method to detect the proliferation of HepG2 cells; while high insulin-induced HepG2 cells to create insulin resistance cell model; impact the detection dihuang oligosaccharides on insulin resistance in HepG2 cells glucose consumption; were used glucose 6 - phosphate dehydrogenase coupled colorimetric lactate dehydrogenase coupled colorimetric ammonium molybdate given P method and anthrone method for the determination of insulin resistance in HepG2 cells glucose kinase (GK), phosphoenolpyruvate acetone acid carboxylase kinase (PEPCK), glucose-6 - phosphatase (G-6-Pase) activity and glycogen content; using RT-PCR technology, detection Rehmannia glutinosa oligosaccharides key gene expression in HepG2 cells to insulin resistance, transcription factor and the regulation of cell differentiation, lipid metabolism, peroxisome proliferator-activated receptor alpha (PPAR-alpha) and insulin receptor (IR); regulation of glucose metabolism in vivo cytokine glucose transporter 4 (GLUT4 ) and glucose transporter 2 (GLUT2). Results: in the concentration of glucose in the medium, the high concentration (of 10 ~~ 30 mg · L -1 ) Dihuang oligosaccharides promote proliferation of HepG2 cells, low concentration (0.1 ~ 3mg · L - 1 ) inhibited the proliferation of HepG2 cells, the effects were significant dose-effect relationship; Rehmannia glutinosa oligosaccharides (0.1 ~ 30mg · L -1 ) can promote glucose consumption in HepG2 cells, the best concentration of 10mg · L -1 ; Rehmannia glutinosa oligosaccharides (0.1 ~ 100mg · L -1 ) can contribute to insulin resistance in HepG2 cells consume glucose, enhanced sensitivity to insulin , against the effect of high insulin-induced insulin resistance and also capable of significantly preventing or reversing the development of insulin resistance in a certain extent. Rehmannia glutinosa oligosaccharides reduces insulin resistance in HepG2 cells glucose - 6 - phosphatase and phosphoenolpyruvate carboxylase kinase activity, enhanced glucokinase activity and increased glycogen content, improving the state of insulin resistance HepG2 cells. Rehmannia glutinosa oligosaccharides can promote insulin resistance in HepG2 cells PPAR-α, IR, GLUT4 mRNA expression; the GLUT2 gene mRNA expression significantly reduced. Conclusion: The high concentration of ROS promote HepG2 proliferation, low concentration, inhibited proliferation; Rehmannia glutinosa oligosaccharides can significantly contribute to insulin resistance in HepG2 cells glucose consumption, to some extent, prevent or reverse the development of insulin resistance; Rehmannia glutinosa oligosaccharides can inhibit insulin resistant HepG2 cells gluconeogenesis, reduce the expression of endogenous glucose production, at the same time accelerate glycolysis conduct, promote cell glucose utilization, enhanced intracellular glycogen content, has significantly improved insulin resistance on HepG2 , which may be related to activation of PPAR-α, to adjust IR HepG2 inside and GLUT2 mRNA expression. The study initially clarified Rehmannia glutinosa oligosaccharides improve hepatic insulin resistance and its molecular mechanism, prevention and treatment of type 2 diabetes provides useful data for Rehmannia glutinosa oligosaccharides.

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