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机构地区:[1]贵州省遵义医学院附属医院肾内科/贵州省细胞工程实验室再生医学学科,遵义市医学硕士研究生563003
出 处:《医学研究生学报》2016年第9期913-917,共5页Journal of Medical Postgraduates
基 金:国家自然科学基金(81260118);遵义医学院博士科研启动基金[(2006)13]
摘 要:目的糖尿病慢性血管病变的组织器官存在不同程度血管新生,由于观察新生血管存在一定困难,文中建立一种便于研究观察的糖尿病体内新生血管生成实验模型。方法 SD大鼠随机分成正常对照组、糖尿病模型组。采用链脲菌素一次性腹腔注射诱导构建SD大鼠糖尿病模型。成模8周后于大鼠腹部皮下植入基质胶(Matrigel),2周后处死大鼠并取出基质胶栓,检测血糖、尿蛋白,HE染色观察新生血管结构,免疫组化及免疫荧光观察血管内皮细胞标志物(JG12)和α-平滑肌肌动蛋白(α-SMA)表达。结果与正常对照组比较,糖尿病模型组大鼠血糖、24 h尿量,24 h尿蛋白均明显增加[(6.15±3.02)mmol/L vs(35.08±6.92)mmol/L,(5.33±1.53)m L vs(105.00±20.82)m L,(7.78±2.12)mg/24 h vs(47.70±9.63)mg/24 h,P<0.01];正常对照组基质胶栓呈白色透明状,而糖尿病模型鼠基质胶栓呈不同程度的红色,可见明显细小血管样结构,HE染色显示正常对照组基质胶栓中偶见血管样结构,而糖尿病模型鼠基质胶栓中见大量的血管样结构,部分管腔内可见大量红细胞。免疫组化、荧光显示糖尿病模型组基质胶栓中JG12及α-SMA表达较正常对照组明显增多[(27.36±3.75)vs(7.76±1.85),(36.85±4.28)vs(10.22±2.64),P<0.01],并呈现血管网样排列。结论高血糖状态可诱生基质胶新生血管,糖尿病大鼠皮下基质胶血管生成模型有助于糖尿病器官组织新生血管生成相关研究。Objective There are different degrees of angiogenesis in the tissues and organs of the patient with diabetic chronic vascular disease, and it is somehow difficult to observe angiogenesis. This study was to construct an experimental model of angiogenesis in the diabetic rat in vivo. Methods Twelve SD rats were equally randomized into a normal control and a diabetes mellitus (DM) group, the DM model constructed by intraperitoneal injection of streptozotocin. At 8 weeks after modeling, matrigel was planted under the abdominal skin of the control and model rats and removed 2 weeks later. Then the structure of the new vessels was observed by HE staining and the expressions of aminopeptidase P (JG12) and alpha-smooth muscle actin (ct-SMA) detected by immunohistochemistry and immunofluorescence. Results Compared with the control rats, the DM models showed significant increases in the blood glucose level ( [ 6.15 ± 3.02 ] vs [ 35.08 ±6.92 ] mmol/L), 24-hour urine volume ( [ 5.33 ± 1.53 ] vs [ 105.00± 20.82 ] mL), and 24-hour urinary protein level ( [ 7.78 ± 2.12 ] vs [ 47.70 ± 9.631 mg 24 h) (P 〈 0.01 ). The matrigel plug was white and transparent in the controls but red with a visible tiny vessel-like structures in the model rats. The vessel-like structures were found occasionally in the matrigel plug of the control animals but in large numbers with many erythrocytes in that of the DM models. In comparison with the normal controls, the model rats exhibited significantly upregulated expressions of JG12 (7.76 ± 1.85 vs 27.36 ± 3.75, P 〈 0.01) and α-SMA (10.22 ±2.64 vs 36.85 ±4.28, P 〈 0.01) in the matrigel plug, and a vessel-like reticular distribution was observed. Conclusion Hyperglycemia may induce angiogenesis in the matrigel plug. The DM rat model of subcutaneous matrigel anglo- genesis may contribute to the study of angiogenesis in diabetes.
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