Caveolin-1对糖尿病大鼠缺血组织中NO的影响及机制

发布时间：2018-12-07 17:20

【摘要】：目的:建立糖尿病下肢缺血大鼠动物模型(STZ缺血大鼠模型),导入质粒转染的小凹蛋白(Caveolin-1),观察Caveolin-1对糖尿病大鼠缺血组织中NO的影响及机制。方法:应用STZ法构建糖尿病大鼠,将80只糖尿病大鼠随机分为4组(每组20只):A组为假手术对照组;B组为缺血组(单纯结扎股动脉);C组为缺血空转染组(结扎股动脉并转染不含Caveolin-1的质粒);D组为缺血转染组(结扎股动脉并转染Caveolin-1),监测各组血糖,并于转染后14天取缺血部位肌肉组织,分别应用Elisa法检测各组组织中NO水平,免疫组织化学染色检测组织中Caveolin-1、e NOS的含量,应用Image-Pro Plus(IPP)6.0图像分析系统对缺血组织中Caveolin-1、e NOS进行半定量分析,HE染色观察组织形态及炎性细胞浸润情况。结果:各组血糖均高于16.8mmol/L,Elisa检测缺血组织中NO,见各组均有NO的表达,缺血组中NO的表达明显低于假手术组(P0.05),缺血转染组组织中的NO水平明显高于其他各组(P0.05)。免疫组化半定量检测显示:缺血组Caveolin-1蛋白的表达较假手术组明显增多(P0.05),e NOS的蛋白水平变化不大(P0.05),NO含量明显减少(P0.05),缺血转染组中的Caveolin-1、e NOS及NO的含量比其他三组明显增多(P0.05)。HE染色见假手术对照组缺血组织细胞形态大致正常,无充血、水肿,缺血组组及空转染组的腓肠肌细胞中均有多量炎症细胞浸润,且肌肉组织萎缩。转染组炎症细胞侵润较少,肌肉萎缩不明显。结论:1、在糖尿病下肢缺血模型中Caveolin-1会低浓度增加,并通过抑制e NOS/NO通路,降低NO的生成2、对糖尿病缺血大鼠人工转染Caveolin-1使其高浓度表达时会促进e NOS/NO通路,使NO生成增加。
[Abstract]:Aim: to establish a rat model of diabetic lower limb ischemia (STZ ischemia rat model) and introduce plasmid transfected concave protein (Caveolin-1) to observe the effect and mechanism of Caveolin-1 on NO in ischemic tissue of diabetic rats. Methods: STZ method was used to construct diabetic rats. 80 diabetic rats were randomly divided into 4 groups (20 rats in each): A group as sham-operation control group, B group as ischemia group (simple ligation of femoral artery), 20 rats in each group as sham-operation control group). Group C was treated with ischemia and empty transfection (ligation of femoral artery and transfection of plasmids without Caveolin-1). Group D was treated with ischemia transfection (ligation of femoral artery and transfection of Caveolin-1). Blood glucose was monitored in each group. Muscle tissue of ischemic area was taken 14 days after transfection. NO level in each group was detected by Elisa method. Immunohistochemical staining was used to detect the content of Caveolin-1,e NOS in tissues, and Image-Pro Plus (IPP) 6.0 image analysis system was used to analyze Caveolin-1,e NOS in ischemic tissues. Tissue morphology and inflammatory cell infiltration were observed by HE staining. Results: the expression of NO in ischemic tissues was detected by Elisa with blood glucose higher than 16.8 mmol / L in all groups. The expression of NO in ischemic group was significantly lower than that in sham operation group (P0.05), and the expression of NO in ischemic group was significantly lower than that in sham operation group (P0.05). The level of NO in ischemic transfection group was significantly higher than that in other groups (P0.05). Immunohistochemical semi-quantitative analysis showed that the expression of Caveolin-1 protein in ischemic group was significantly higher than that in sham operation group (P0.05), e NOS protein level was not changed (P0.05), NO content was significantly decreased (P0.05). The contents of Caveolin-1,e NOS and NO in the ischemic transfection group were significantly higher than those in the other three groups (P0.05). HE staining showed that the morphology of ischemic tissue cells in the sham-operation control group was approximately normal without hyperemia and edema. Many inflammatory cells were infiltrated in gastrocnemius muscle cells in ischemia group and empty transfection group, and muscle tissue atrophy. In the transfection group, the inflammatory cells invaded less and the muscle atrophy was not obvious. Conclusion: 1. In the diabetic model of lower extremity ischemia, the low concentration of Caveolin-1 was increased, and the formation of NO was decreased by inhibiting the e NOS/NO pathway. Artificial transfection of Caveolin-1 into diabetic ischemic rats promoted e NOS/NO pathway and increased NO production.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R587.2

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