低氧低温对大鼠骨骼肌细胞脂代谢关键酶及解耦联蛋白1的影响

Effects of hypoxia and hypothermia on key enzymes of lipid metabolism and uncoupling protein 1 in skeletal muscle cells of rat

日的观察低氧低温对大鼠骨骼肌细胞脂代谢关键酶及解耦联蛋白1的影响方法分离骨骼肌,培养骨骼肌细胞,观察细胞形态并鉴定细胞,随机分为对照组、低氧组、低温组、低氧低温组,处理48 h后通过RT-qPCR及Westem-Blot实验检测脂肪分解关键酶〔激素敏感性甘油三酯脂肪酶(HSL)、肉碱棕榈酰转移酶1A(CPT1A)〕及棕色脂肪标志性解耦联蛋白1(UCP1)的mRNA及蛋白表达水平。结果(1)RT-qPCR结果显示,与对照组、低氧组、低温组比,低氧低温组HSL mRNA表达增多(P<0.05);与低温组比,低氧低温组CPT1A mRNA表达减少(P<0.05)。与对照组比,低氧低温组UCP1 mRNA表达增多(P<0.05)。(2)Western-Blot结果显示,与对照组、低氧组、低温组比,低氧低温组HSL蛋白表达增多(P<0.05)。与对照组、低氧组比,低氧低温组CPTIA蛋白表达减少(P<0.05),与低温组相比增多(P<0.05)。与对照组比,三组UCP1蛋白表达减少(P<0.05)。结论低氧低温对骨骼肌细胞的脂代谢水平有一定的影响,促进脂代谢关键酶HSL mRNA及蛋白表达,抑制CPT1A mRNA及蛋白表达,促进骨骼肌细胞UCP1 mRNA表达,抑制骨骼肌细胞UCP1蛋白表达。

To observe the effects of hypoxia and hypothermia on key enzymes of lipid metabolism and uncoupling protein 1 in skeletal muscle cells of rat.Methods Skeletal muscle cells were isolated,cultured,observed and identified.They were randomly divided into control group,hypoxia group,hypothermia group,hypoxia and hypothermia group.The mRNA and protein expression levels of hormone sensitive triglyceride lipase(HSL),carnitine palmitoyltransferase 1 A(CPTIA),uncoupling protein 1(UCP1)of brown fat marker were detected by RT-qPCR and Western-Blot after 48 hours of treatment.Results(1)The results of RT-qPCR showed that compared with the control group,hypoxia group,hypothermia group,the expression of HSL mRNA in hypoxia and hypothermia group was increased(P<0.05);compared with the hypothermia group,the expression of CPT1 A mRNA decreased in hypoxia and hypothermia group(P<0.05).Compared with the control group,UCP1 mRNA increased in the hypoxia and hypothermia group(P<0.05).(2)The results of Western-Blot showed that compared with the control group,hypoxia group,and hypothermia group,the expression of HSL protein increased in hypoxia and hypothermia group(P<0.05).Compared with the control group and hypoxia group,the expression of CPT1A protein in the hypoxia and hypothermia group was decreased(P<0.05),while that of the hypothermia group increased(P<0.05).Compared with the control group,the expression of UCP1 protein in all the other three groups were decreased(P<0.05).Conclusion Hypoxia and hypothermia have certain effects on the level of lipid metabolism of skeletal muscle cells,which promote the expression of HSL mRNA and protein,inhibit the expression of CPT1A mRNA and protein.Also,it could promote the expression of UCP1 mRNA and inhibit the expression of UCP1 protein.