波生坦对糖尿病小鼠血管病变的保护作用

Protective effect of bosentan on diabetic vascular disease in rats

目的研究内皮素受体拮抗剂波生坦对糖尿病小鼠血管病变的保护作用及其机制。方法 60只雄性C57BL/6小鼠随机分为正常对照组、糖尿病对照组、糖尿病药物干预组。链脲佐菌素(streptozotocin,STZ)(70 mg/kg)腹腔注射建立糖尿病模型,尾静脉空腹血糖测定检测建模是否成功。模型建立后,糖尿病药物干预组给予波生坦(100 mg/kg)灌胃,1次/d,连续8周;正常对照组、糖尿病对照组给予同等剂量0.9%氯化钠注射液灌胃8周。8周后观察各组主动脉HE染色情况、主动脉实时定量PCR检测血管内皮生长因子(vascular endothelial growth factor,VEGF)、内皮素1(endothelin-1,ET-1)、肿瘤坏死因子α(tumor necrosis factorα,TNF-α)、基质金属蛋白酶2(matrix metalloproteinases 2,MMP-2)、基质金属蛋白酶9(matrix metalloproteinases 9,MMP-9)的m RNA表达水平。结果主动脉HE染色示对照组大致正常;糖尿病组主动脉斑块形成明显,大量炎症细胞浸润;糖尿病药物干预组斑块负荷明显减轻,炎症细胞浸润减少。主动脉实时定量PCR检测结果示糖尿病组与糖尿病药物干预组VEGF m RNA表达低于正常对照组(P<0.05),ET-1、TNF-α、MMP-2、MMP-9 m RNA表达高于正常对照组(P<0.05);糖尿病药物干预组VEGF m RNA表达较糖尿病组升高(P<0.05);糖尿病药物干预组ET-1、TNF-α、MMP-2、MMP-9 m RNA表达较糖尿病组降低(P<0.05)。结论内皮素受体拮抗剂波生坦对糖尿病小鼠主动脉病变有保护作用。

Objective To investigate the potential beneficial effects of endothelin receptor antagonist bosentan on vascular disease in diabetic mice and its mechanism. Methods Sixty 6-week-old male C57BL/6 mice were divided into 3 groups： control group, diabetes group and diabetes with drug intervention group. Streptozotocin（STZ） was intraperitoneally injected at a single dose of 70 mg/kg to establish diabetes model. The fasting blood glucose from tail vein sample was tested to verify diabetes model. Bosentan（100 mg/kg） was given by intragastric administration once a day after STZ injection for 8 weeks in diabetes with drug intervention group, and the same amount of 0.9% sodium chloride injection was given to control group and diabetes group. After 8 weeks, aortic HE staining results in each group and m RNA expression of vascular endothelial growth factor（VEGF）, endothelin-1（ET-1）, tumor necrosis factorα（TNF-α）, matrix metalloproteinases 2（MMP-2）, matrix metalloproteinases 9（MMP-9） were observed by q RTPCR. Results HE staining results showed that aortic intima was generally normal in control group, while there were atherosclerotic plaque formation and a large number of inflammatory cell in diabetes group and the plaque burden lightened obviously in diabetes with drug intervention group. q RT-PCR results showed that the m RNA level of VEGF in diabetes group and diabetes with drug intervention group were significantly lower than those in control group（P〈0.05）, while the m RNA expression of ET-1, TNF-a, MMP-2, MMP-9 in diabetic group and diabetes with drug intervention group increased significantly when compared with control group（P〈0.05）. The m RNA expression of VEGF in diabetes with drug intervention group was higher than that in diabetes group（P〈0.05） and the m RNA expressions of ET-1, TNF-a, MMP-2 and MMP-9 in diabetes with drug intervention group were lower than those in diabetic group（P〈0.05）. Conclusion Endothelin receptor antagonist bosentan has beneficial effects on vascular complications in diabetic mice.