苏木及巴西苏木素调控自噬对高糖诱导血管内皮细胞损伤的影响

发布时间：2018-09-14 13:10

【摘要】：目的探讨中药苏木(Caesalpinia Sapan L)及其主要有效成分巴西苏木素(Brazilin)对高糖诱导血管内皮细胞损伤的保护作用及其机制,为其治疗糖尿病大血管病变提供实验依据。方法动物实验:30只健康雄性SD大鼠随机分为正常组(n=10)与造模组(n=20),正常组正常饲养,造模组高脂高糖饲料连续喂养4周后,一次性尾静脉注射链脲佐菌素(STZ)溶液造成糖尿病模型,成模后的大鼠随机分为对照组与苏木组,苏木组每天按人等效剂量810mg/kg体重生药量,灌服溶于2ml蒸馏水中的苏木颗粒剂,正常组与对照组每天灌服等量的生理盐水,以上大鼠4周后测血糖及体重,取主动脉,运用HE染色观察糖尿病大鼠主动脉形态结构;透射电镜(tem)观察糖尿病大鼠血管内皮细胞中的自噬结构,探讨苏木对糖尿病大鼠血管内皮细胞自噬的影响。细胞实验:培养人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC),采用MTT法确定巴西苏木素浓度;加入浓度为33.3 mmol/l的葡萄糖溶液,建立高糖诱导HUVEC损伤体外模型,给予浓度为15 ug/ml巴西苏木素,Western-blot检测自噬相关基因Beclin1、LC3-Ⅱ、p62蛋白的表达。结果动物实验:糖尿病造模成功的共有18只,再随机将18只分为对照组(n=9)和苏木组(n=9),所有存活大鼠均毛色、活动正常,造模组体重减轻。血糖较造模前增加(P0.05),造模组大鼠体重低于正常组,造模组中苏木组体重低于对照组(P0.05)。HE染色观察大鼠主动脉形态结构,苏木组和对照组,较正常组血管平滑肌均有损伤,但苏木组较对照组,内膜虽有破坏,却已有改善。透射电镜观察糖尿病大鼠血管内皮细胞中的自噬结构,苏木组较正常组,胞浆内自噬体数目增多、体积变大,线粒体呈嵴紊乱或空泡化。对照组糖尿病大鼠血管中的自噬体数量较正常组减少。细胞实验:MTT实验选定用于实验的巴西苏木素浓度为15 ug/ml;Beclin1、LC3-Ⅱ的蛋白表达,高糖处理组较正常组低,巴西苏木素组较高糖处理组高(P0.05)。p62的蛋白表达,高糖处理组较正常组高,巴西苏木素组低于正常组和高糖处理组(P0.05)。结论中药苏木及其主要有效成分巴西苏木素通过调控自噬而对高糖诱导血管内皮细胞发挥保护作用,为治疗糖尿病大血管病变提供实验依据。
[Abstract]:Objective To investigate the protective effect and mechanism of Caesalpinia Sapan L and its main active component Brazilin on vascular endothelial cell injury induced by high glucose, and to provide experimental basis for its treatment of diabetic macroangiopathy. (n=20), the normal group was fed normally, and the model group was fed with high fat and high sugar diet for 4 weeks. The diabetic rats were randomly divided into control group and hematoxylin group. The hematoxylin group was fed with hematoxylin granules dissolved in 2 ml distilled water at 810 mg/kg body weight per day. The blood glucose and body weight were measured after 4 weeks. The aorta of diabetic rats were stained with HE. The autophagy structure of vascular endothelial cells in diabetic rats was observed by transmission electron microscopy (tem). Effects. Cell experiment: Cultured human umbilical vein endothelial cells (HUVEC), determined the concentration of Brazilian hematoxylin by MTT method; added glucose solution with a concentration of 33.3 mmol/l, established high glucose-induced HUVEC injury in vitro model, given concentration of 15 ug/ml Brazilian hematoxylin, Western-blot detection of autophagy-related genes. Results Animal experiment: 18 diabetes mellitus rats were successfully established, and then randomly divided into control group (n=9) and hematoxylin group (n=9). All the surviving rats were hair color, normal activity, weight loss of model group. Blood glucose increased (P 0.05), body weight of model group was lower than that of normal group. HE staining was used to observe the morphology and structure of aorta in diabetic rats (P 0.05). The vascular smooth muscle of hematoxylin group and control group were damaged, but the intima of hematoxylin group was damaged, but it was improved. The autophagy structure of vascular endothelial cells in diabetic rats was observed by transmission electron microscope. The number of autophages in the blood vessels of diabetic rats in the control group was lower than that of the normal group. Cell experiment: MTT assay selected Brazilian hematoxylin concentration for the experiment was 15 ug/ml; Beclin1, LC3-II protein expression, high glucose treatment group was lower than the normal group, Brazilian hematoxylin group was higher. The expression of p62 protein was higher in the sugar treatment group than in the normal group, and lower in the Brazilian hematoxylin group than in the normal group and the high glucose treatment group (P 0.05). Conclusion The Chinese herbal hematoxylin and its main effective component Brazilian hematoxylin can protect the vascular endothelial cells induced by high glucose by regulating autophagy, and can be used to treat diabetic macroangiopathy. For experimental basis.
【学位授予单位】：锦州医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R587.2

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