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1,25-二羟维生素D3对实验糖尿病大鼠模型肝脏中甘油三酯含量的影响及其机制探讨

Investigation of the mechanism of 1,25-dihydroxyvitamin D3 on triglycerides content in liver of experimental diabetic rat model

摘要:

目的 活性维生素D与2型糖尿病、高脂血症的发生发展密切相关,通过观察1,25-二羟维生素D3[1,25(OH)2D3]对实验糖尿病大鼠模型肝脏中过氧化物酶增殖物激活受体-α(PPAR-α)、肝细胞核因子-4α(HNF-4α)、胆固醇调节元件结合蛋白-1C(SREBP-1C)、载脂蛋白A5(apoA5)表达及甘油三酯含量的影响,探讨1,25(OH)2D3调控肝脏甘油三酯的机制.方法 取80只健康雄性Wistar大鼠,分成糖尿病组20只、1,25(OH)2D3干预组(VitD干预组)20只、PPAR-α抑制组20只和正常对照组20只.以随机数字表法,取各组大鼠各15只,心内取血检测生化指标,采用实时荧光定量PCR、Western印迹法分别对大鼠的肝脏组织中PPAR-α、HNF-4α、SREBP-1C、apoA5基因及蛋白表达水平进行检测,并用酶法检测肝脏组织中甘油三酯含量.结果 (1)糖尿病组肝脏中PPAR-α、HNF-4α、apoA5基因、蛋白表达较正常对照组显著降低(P<0.05),而SREBP-1C基因、蛋白表达及甘油三酯含量较正常对照组显著增加(P<0.05).(2)VitD干预组肝脏中PPAR-α、HNF-4α、apoA5基因及蛋白表达较糖尿病组明显增加(P<0.05),而SREBP-1C基因及蛋白、甘油三酯含量较糖尿病组明显减少(P<0.05).(3)VitD干预组肝脏中PPAR-α、HNF-4α、SREBP-1C、apoA5基因、蛋白表达、甘油三酯含量较正常对照组无显著性差异(P>0.05).(4)PPAR-α抑制组PPAR-α、HNF-4α、apoA5基因、蛋白表达较正常对照组显著降低(P<0.05),而SREBP-1C基因、蛋白表达、甘油三酯含量较正常对照组明显增加(P<0.05).(5) PPAR-α抑制组PPAR-α基因、蛋白表达较糖尿病组显著降低(P<0.05),而HNF-4α、apoA5、SREBP-1C基因、蛋白表达及甘油三酯含量与糖尿病组无显著性差异(P<0.05).(6) PPAR-α抑制组PPAR-α、HNF-4α、apoA5基因、蛋白表达较VitD干预组显著降低(P<0.05),而SREBP-1C基因、蛋白表达及甘油三酯含量较VitD干预组显著增加(P<0.05).结论 1,25(OH)2D3可能通过上调肝脏中PPAR-α、HNF-4α、apoA5表达,抑制SREBP-1C的表达,从而降低甘油三酯水平.

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abstracts:

Objective Activity of vitamin D is correlated with type 2 diabetes mellitus and hyperlipidemia,but the concrete relationship of 1,25-dihydroxyvitamin D3 [1,25 (OH) 2 D3] and triglyceride is unknown.This study was to observe the gene and protein expression of peroxisome proliferator-activated receptor α(PPAR-α),hepatocyte neclear factor-4α (HNF-4α),sterol regulatory element binding proteins-1C (SREBP-1 C),apolipoprotein (apoA5) during 1,25 (OH)2D3 regulation of triglyceride in liver of diabetes rat,as well as to investigate the mechanism of 1,25 (OH) 2D3 in influencing triglyceride in liver.Methods Eighty male Wistar Rats were divided into 4 groups:20 rats with diabetes mellitus(DM group),20 rats of 1,25 (OH)2D3 treated group (V itD group),20 rats with PPAR-α inhibition group,and 20 rats of normal control group.15 from each group were taken in random by random number table to detect biochemical indicators and to detect the gene and protein expressions of PPAR-α,HNF-4α,SREBP-1 C,apoA5 in liver,using realtime-fluorescent quantitation reverse transcription,polymerase chain reaction and Western blot.The level of triglyceride in liver was also detected.Results (1) The gene and protein expression of PPAR-α,HNF-4α,apoA5 in liver of diabetic rat was lower than that in normal control group (P < 0.05).The expression of SREBP-1C and the level of triglyceride in liver of diabetic rat were higher than those of normal control group (P<0.05).(2)The expression of PPAR-α,HNF-4α,apoA5 in liver of VitD group were higher than those of DM group(P<0.05).The gene and protein expression of SREBP-1C and the level of triglyceride in VitD group were lower than those in DM group(P<0.05).(3) The expression of PPAR-α,HNF-4α,SREBP-1 C,apoA5,and the level of triglyceride showed no significant difference between VitD group and normal control group(P>0.05).(4) The expression of PPAR-α,HNF-4α,and apoA5 in liver of PPAR-α inhibition group was lower than that in normal control group(P<0.05).The expression of SREBP-1C and the level of triglyceride in liver of PPAR-α inhibition group were higher than those in normal control group(P<0.05).(5) The expression of PPAR-α in liver of PPAR-α inhibition group was lower than that in diabetic rat (P<0.05).The expression of HNF-4α,apoA5,SREBP-1 C,and the level of triglyceride showed no significant difference between PPAR-α inhibition group and diabetic rat(P>0.05).(6) The expression of PPAR-α,HNF-4α,apoA5 in liver of PPAR-α inhibition group was lower than that in VitD group(P< 0.05).The expression of SREBP-1C and the level of triglyceride in liver of PPAR-α inhibition group were higher than those in VitD group (P < 0.05).Conclusions 1,25 (OH) 2D3 may lower the level of serum triglyceride by upregulating the expression of PPAR-α,HNF-4α,and apoA5 and inhibiting the expression of SREBP-1C in liver.

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作者: 阳琰 [1] 高琳 [1] 晏永慧 [1] 王小英 [1] 廖鑫 [1] 黄琦 [1] 张晗 [1] 陈其容 [1]
期刊: 《中华内分泌代谢杂志》2014年30卷12期 1115-1119页 ISTICPKUCSCD
栏目名称: 基础研究
DOI: 10.3760/cma.j.issn.1000-6699.2014.12.016
发布时间: 2015-01-21
基金项目:
国家自然科学基金 贵州省科学技术基金项目 贵州省优秀科技教育人才省长专项资金项目 贵州省科技合作项目
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