摘要
目的观察自发性Ⅱ型糖尿病OLETF大鼠糖尿病模型出现的时间,罗格列酮对过氧化物酶体增殖体激活受体(PPAR)γ基因表达的干预作用。方法OLETF大鼠随机分成糖尿病对照组和罗格列酮干预组,8周龄时,干预组以罗格列酮每日3mg/kg体重灌胃,直至40周龄。行口服葡萄糖耐量试验(OGTT),鉴定糖尿病发病情况。应用TaqMan实时荧光定量逆转录聚合酶链反应(RTPCR)技术分析PPARγmRNA在糖尿病大鼠各组织中的表达水平及罗格列酮对PPARγ基因的影响。结果至40周龄,对照组糖尿病累积发病率92.5%,糖耐量异常发生率7.5%。干预组糖尿病的累计发病率仅为28.6%,糖耐量异常发生率7.5%,显著低于对照组(P<0.01)。实时荧光定量结果表明:PPARγ在大鼠各组织中均有分布,对照组大鼠各组织中以脂肪表达量最高2.71±0.14(单位:1010拷贝数/100mg组织),是其他组织的10～100倍;其次为血管、胰腺、肾脏、肺脏,分别为1.15±0.10、2.58±0.064、1.52±0.12、4.67±0.088(单位:109拷贝数/100mg组织);心、肝、脾、肌肉中的拷贝数最低分别为7.77±0.11、4.31±0.12、1.51±0.21、2.70±0.087(单位:108拷贝数/100mg组织);干预组各组织中PPARγmRNA的表达量均比对照组高,且在血管、胰腺、肌肉、肾脏组织中差异有统计学意义(P<0.01)。
Objective To observe the effect of rosiglitazone intervention in spontaneous diabetic OLETF rats and develop a method for absolute quantification of peroxisome proliferator-activated receptor gamma (PPARγ) mRNA by using real-time polymerase chain reaction (PCR) in tissues.Methods Thirty OLETF male rats with 4 weeks age were divided into 2 groups randomly:diabetic control group (n= 15) and rosiglitazone intervention group (n= 15).The intervention group was administered with rosiglitazone (3 mg/kg) by gavage daily from 8 weeks old to 40 weeks old.The incidence of rats with DM and IGT was determined by OGTT.PPARγ mRNA was detected by absolute quantification RT-PCR and the effect of rosiglitazone intervention was abserved.Results On the week 40,the cumulative incidences of control group with DM or IGT were 92.5% (13/14) or 7.5% (1/14),respectively,while that in intervention group with DM was 28.6%(1/15).PPARγ mRNA was present in all the tissues,with the greatest expression in adipose tissue 2.71± 0.14 (10~ 10copies/100 mg tissue),with values 10-100 times more than that in other tissues.High level of PPARγ was detected in aorta,pancreas,kidney,lung, 1.15± 0.10, 2.58± 0.064, 1.52± 0.12, 4.67± 0.088 (10~9 copies/100 mg tissue),respectively,while on the other hand,only very low levels were detected in skeletal muscle,heart,liver and spleen, 7.77± 0.11, 4.31± 0.12, 1.51± 0.21, 2.70± 0.087 (10~8 copies/100 mg tissue),respectively.Statistical analysis indicated that PPARγ mRNA expression was significantly increased in intervention group,and in vessel,pancreas,skeletal muscle and kidney tissues,respectively (P< 0.05).Conclusion Rat diabetic model was established successfully.Rosiglitazone could increase the expression of PPARγ mRNA in main tissues of diabetic rats.It suggests that the prevention of rosiglitazone on OLETF rats may be associated with the increase of PPARγ mRNA.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2005年第6期746-749,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目(10703H52)
