摘要
目的:了解抗氧化剂α-硫辛酸对己糖胺通路的限速酶谷氨酰胺:6-磷酸果糖转酰胺酶(glutamine: fructose-6-phosphate amidotransferase, GAFT)基因表达的影响,为进一步研究抗氧化剂对糖尿病的治疗提供理论依据和实验基础。
方法:1.应用四氧嘧啶制备糖尿病小鼠模型。
2.将糖尿病小鼠分层随机分成四组,分别腹腔注射安慰剂或不同剂量(15、30、60mg/kg体重)α-硫辛酸14天。
3.使用TBA法检测小鼠血清和后肢骨骼肌中丙二醛(malondialdehyde, MDA)含量。
4.提取小鼠后肢骨骼肌中的总RNA,采用半定量RT-PCR的方法测定GFAT1mRNA的辉度。
结果:1.四氧嘧啶制备糖尿病小鼠模型成功率69.2%。
2.安慰剂组血清、骨骼肌组织中MDA含量最高(P<0.05):正常对照组与高剂量组、中剂量组血清MDA含量未见差异(P>0.05),低剂量组高于正常对照组(P<0.05):低、中、高剂量三组小鼠骨骼肌组织中MDA含量随α-硫辛酸用量的增加而减少(P<0.05),高剂量组与正常对照组未见差异(P>0.05)。
3.安慰剂组骨骼肌GFAT1mRNA的辉度高于正常对照组、中、高剂量组(P<0.05),与低剂量组间未见差异(P>0.05),但数值高于低剂量组;中、高剂量组间未见差异(P>0.05),但高于正常对照组(P<0.05)。
4.骨骼肌组织中MDA含量与GFAT1mRNA的辉度成正相关(P<0.05)。
5.高、中、低剂量组小鼠给药前后体重均未见差异(P>0.05),血糖较给药前下降(P<0.05);正常对照组体重增加(P<0.05)。
结论:糖尿病动物体内MDA含量和GFAT1mRNA表达增加,给予抗氧化
Objective:To learn the effect of alpha-lipoic acid on the expression of glutamine: fructose-6-phosphate amidotransferase 1, which is the rate-limiting enzyme of hexosamine biosynthesis pathway, in the diabetic mouse hind-limb skeletal muscle, and provide the theory basis for applying the antioxidant alpha-lipoic acid in preventing and treating diabetes. Methods:1. Diabetes was induced with a single intravenous injection of alloxan into KM mice.2. Diabetic mice were divided into four groups at random. The placebo (30mg/kg ip) or different doses alpha-lipoic acid (15,30,60mg/kg ip) was administered daily to these mice for fourteen days.3. Malondialdenhyde levels serve as a marker for lipid peroxidation. These were measured using thiobarbituric acid in the serum and the skeletal muscle among normal and diabetic mice.4. The expression of glutamine: fructose-6-phosphate amidotransfer -ase 1 gene was analyzed by semi-quantitative reverse transcrip -tion polymerase chain reaction.Result:1. The incidence of diabetes induced by alloxan was 69.2%.2. The placebo group's MDA content in the serum and the skeletal muscle was the highest among groups (P<0.05). In the serum, the second was the low dosage group (P<0.05), and there was no significantly different among normal, medium dosage, and high dosage (P>0.05). The skeletal muscle's MDA content reduced with the increase of alpha-lipoic acid among diabetic mice (P<0.05), but between high dosage group and normal group there was no obviously different (P>0.05).3. In the skeletal, the placebo group's glutamine: fructose-6 -phosphate amidotransferase 1 mRNA level is as more as the low dosage group's (P>0.05), however, it's mean was higher than the low dosage group's. The two groups were higher than the medium and high dosage groups (P<0.05), and between the later two groups there was no significantly
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