摘要
目的研究尾静脉注射人胰高血糖素样肽-1(hGLP-1)类似物基因(2×Val2-hGLP-1)重组表达质粒对糖尿病模型大鼠血糖、血清胰岛素水平及胰岛细胞的影响。方法建立链脲佐菌素(STZ)诱导SD糖尿病大鼠模型,随机分为含有hGLP-1类似物基因的重组质粒(pIRES2-EGFP/Val2-hGLP-1)转染组、空质粒(pIRES2-EGFP)转染组、糖尿病模型对照组,每组8只。另取8只未经处理的SD大鼠作为正常对照组。重组质粒转染组和空质粒转染组分别通过尾静脉注射含相应质粒(110μg/只)的溶液,糖尿病模型对照组和正常对照组给予等量的生理盐水。实验动物共干预30d,实验结束后,观察各组大鼠血糖及血清胰岛素水平的变化,采用糖耐量及胰岛素耐量实验检测大鼠胰岛素敏感性然后处死动物,HE染色观察胰岛细胞病变情况,免疫组化法分析胰岛素分泌情况。结果与正常对照组、糖尿病模型对照组和空质粒转染组比较,重组质粒转染组血糖水平明显降低(P<0.01),血清胰岛素水平显著升高(P<0.01)。基因治疗改善了糖尿病大鼠的糖耐量,降低了胰岛素抵抗,提高了机体对胰岛素的敏感性(P<0.01)。空质粒转染组与糖尿病模型对照组比较,以上指标的差异均无统计学意义(P>0.05)。同时,重组质粒转染组的胰岛素分泌水平提高,与糖尿病模型对照组比较,胰岛细胞病变程度明显改善。结论hGLP-1类似物基因转染可促进胰岛细胞增殖,提高胰岛素敏感性,从而显著降低糖尿病大鼠的血糖水平,并改善胰岛组织病变程度。
Objective To explore the effects of recombinant expression plasmid of human glucagon-like peptide-1 （hGLP-1） analog gene （2×Val2-hGLP-1） on blood glucose, serum insulin level and pancreatic island in diabetic rats. Methods The diabetic rat model was reproduced by intraperitoneal injection of streptozotocin. The rats were then divided into 3 groups randomly （8 each）： recombinant plasmid pIRES2-EGFP/Val2-hGLP-1 transfection group, empty plasmid pIRES2-EGFP transfection group, and diabetic rat model control group. Moreover, 8 untreated SD rats were set as normal control. Each rat in empty plasmid transfection group and recombinant plasmid transfection group was injected via tail vein with 110μg plasmid, whibe those in diabetic model control group and normal control group were treated with equal volume of normal saline solution. Blood glucose and serum insulin levels of rats were determined 30 days after experiment, and glucose tolerance test and insulin tolerance test were performed to estimate insulin sensitivity. The pathological changes in pancreatic island and insulin secretion were evaluated with HE and immunohistochemistry staining, respectively. Results Compare with normal group, diabetic model group and empty plasmid transfection group, the blood glucose level significantly lowered （P0.01） and the serum insulin levels markedly elevated （P0.01） in recombinant plasmid transfection group. Furthermore, hGLP-1 analog gene therapy improved glucose tolerance, attenuated insulin resistance, and enhanced sensitivity to insulin （P0.01）. No statistically significant difference of these parameters was found between empty plasmid transfection group and diabetic model control group （P0.05）. Meanwhile, the insulin secretion was increased and the pathological changes in pancreatic islands were alleviated in recombinant plasmid transfection group compared with that in diabetic model control group. Conclusions hGLP-1 analog gene transfection may be able to promote the proliferation of pancreatic islands and enhance sensitivity to insulin, thus significantly lower blood glucose level and ameliorate the lesion of pancreatic islets in diabetic rats.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2010年第3期260-263,共4页
Medical Journal of Chinese People's Liberation Army
基金
广东省自然科学基金(7009923)
广东省科技计划项目(2008B030301031)
东莞市高等院校科研机构科技计划项目(2008108101030)
湛江市科技招标项目([2008]74号)
关键词
糖尿病
基因
hGLP-1类似物
转染
葡萄糖耐量试验
胰岛
diabetes mellitus
genes
hGLP-1 analog
transfection
glucose tolerance test
islets of Langerhans
