益气养阴活血方对糖尿病大鼠血清内皮脂肪酶EL水平的影响

发布时间：2018-11-25 09:17

【摘要】：目的：该实验通过腹腔注射链脲佐菌素（STZ）造糖尿病大鼠模型，并应用益气养阴活血方对成模的糖尿病大鼠进行治疗，观察该药对糖尿病大鼠的血糖、血脂、内皮脂肪酶（EL）和大鼠胸主动脉内皮组织形态的影响，研究本方对成模糖尿病大鼠大血管病变的治疗作用及可能的致病机制。方法：选用雄性的清洁级wistar大鼠50只，体重230g～250g，普通饲料适应性喂养4天后，采用完全随机法将50只大鼠分为5组：正常组（N）6只，模型组（M）、中药组（益气养阴活血方组，Z）、西药组（辛伐他汀组，X）、中药+西药组（益气养阴活血方+辛伐他汀组，ZX）各11只。参考相关文献，M、Z、X、ZX组大鼠给予浓度为1%的链脲佐菌素（STZ）柠檬酸缓冲液（柠檬酸缓冲液浓度为0.1mmol/L，pH4.5）按60mg/kg体重一次性腹腔注射，N组则给予相应体积的1%柠檬酸-柠檬酸钠溶液腹腔注射。72h后大鼠尾静脉采血测血糖，若大鼠血糖超过16.7mmol/L，饮水量及尿量均显著增多，则造模成功。Z组给予益气养阴活血方按3.17ml/kg体重水煎浓缩液灌胃，X组给予辛伐他汀片浓缩液0.33mg/kg灌胃，ZX组上午给予与中药组等量益气养阴活血中药，下午给予与西药组等量辛伐他汀灌胃，N组与M组给予相应中药体积的生理盐水灌胃，每日一次，共灌胃12周。于末次灌胃后禁食、禁水12h，2%戊巴比妥钠按20mg/kg体重腹腔注射麻醉，断头取血，检测血浆葡萄糖、总胆固醇、低密度脂蛋白胆固醇及高密度脂蛋白胆固醇。无菌操作取胸主动脉1-2cm作为标本，，置于4%的多聚甲醛溶液中固定，制作切片行HE染色，观察血管内皮组织变化情况，酶联免疫吸附法检测血清内皮脂肪酶（EL）的水平。结果： 1各组大鼠的一般情况在实验过程中，N组大鼠精神饱满，活动自如，饮食、饮水、尿量均正常。M组大鼠则出现精神萎靡，明显多饮、多食、多尿，体重逐渐减轻，毛色枯黄，反应迟钝等表现，符合糖尿病临床“三多一少”的特征。各治疗组大鼠表现介于正常组与模型组之间。偶有大鼠出现肢体活动不利、皮肤溃烂等表现。M组、ZX组各死亡2只，Z组死亡1只。 2各组大鼠血糖水平的观察 M组及各治疗组大鼠血糖均增高，与正常组相比，差别有统计学意义（P0.05）；X组与M组之间比较血糖变化不明显，无统计学意义（P0.05）；Z及ZX组与M组之间比较血糖降低，差别有统计学意义（P0.05）；Z组与ZX组之间比较血糖变化不明显，无统计学意义（P0.05）。 3各组大鼠总胆固醇（TC）、高密度脂蛋白胆固醇（HDL）、低密度脂蛋白胆固醇（LDL）水平的观察 M组及各治疗组大鼠TC均增高，与正常组比较，差别有统计学意义（P0.05）；Z组与M组之间比较TC变化不明显，无统计学意义（P0.05）；X组及ZX组与M组之间比较TC降低，差别有统计学意义（P0.05）；X组与ZX组之间比较TC变化不明显，无统计学意义（P0.05）。 M组及各治疗组大鼠LDL均增高，与正常组比较，差别有统计学意义（P0.05）；Z组与M组之间比较LDL变化不明显，无统计学意义（P0.05）；X组及ZX组与M组之间比较LDL降低，差别有统计学意义（P0.05）；ZX组与X组比较，LDL降低，差别有统计学意义（P0.05）。 M组及各治疗组与正常组大鼠之间比较，HDL明显降低，差别有统计学意义（P0.05）；各治疗组与M组之间比较HDL明显增高，差别有统计学意义（P0.05）；三个治疗组之间两两比较，HDL有明显变化，差别有统计学意义（P0.05）。 4各组大鼠胸主动脉内皮组织形态的观察 HE染色后N组内皮细胞纹理清晰，走形正常，细胞浆着色均匀，胸主动脉内皮细胞扁平，内膜光滑、平坦，中膜是平行相间排列的平滑肌细胞（smooth muscle cell，SMC）和弹力板。M组大鼠胸主动脉内膜呈局限性增厚且局部隆起，近内膜的中膜平滑肌细胞可见增生及排列紊乱，增生平滑肌细胞处的弹力板排列不规则，呈波浪状，偶可见弹力板断裂、分离现象。各治疗组大鼠胸主动脉内皮细胞的组织形态变化较模型组明显减轻。 5各组大鼠酶联免疫吸附法检测血清内皮脂肪酶（EL）表达的情况 M组及各治疗组大鼠EL均增高，与正常组比较，差别有统计学意义（P0.05）；各治疗组与M组之间比较EL明显降低，差别有统计学意义（P0.05）；三个治疗组之间两两比较，EL有变化明显，差别有统计学意义（P0.05）。结论： l益气养阴活血方可使糖尿病大鼠的一般状态好转，降低血糖，调节脂代谢。 2益气养阴活血方可使糖尿病大鼠胸主动脉内皮组织的病理形态好转，有治疗糖尿病大血管内皮病变的作用，可为临床治疗提供治疗方案。 3益气养阴活血方治疗糖尿病大鼠大血管病变的作用可能与其能够降低血糖，改善脂代谢，抑制血清内皮脂肪酶（EL）表达的作用有关。
[Abstract]:Objective: To study the blood glucose and blood lipid of the diabetic rats by means of the experimental study of the model of diabetic rats by intraperitoneal injection of the streptozoin (STZ) and the treatment of diabetic rats with the effects of invigorating qi, nourishing yin and promoting blood circulation. The effects of endothelial lipase (EL) and the morphology of the endothelial tissue of the thoracic aorta in rats were studied, and the therapeutic effect and possible mechanism of the recipe on the large-vessel lesions of the model-type diabetic rats were studied. Methods: 50 rats were randomly divided into 5 groups: normal group (N) 6, model group (M), and traditional Chinese medicine group (qi-nourishing and yin-nourishing and blood-activating group). Z), western medicine group (simvastatin group, X), traditional Chinese medicine + western medicine group (Yiqi Yangyin Huoxue Fang + Simvastatin group, ZX) 1 1. Reference to the relevant literature, M, Z, X, ZX group, rats with a concentration of 1% of a chain-cometacin (STZ) citrate buffer (citric acid buffer at a concentration of 0. 1mmol/ L, pH 4.5) were used as one-time abdominal cavity at 60mg/ kg body weight The blood glucose of the rat tail vein was measured by intraperitoneal injection of the corresponding volume of 1% citric acid-sodium citrate solution, and the blood glucose was measured in the tail vein of the rat after 72h. If the blood sugar in the rat was more than 16.7mmol/ L and the amount of water and urine increased, the model was made. In the group of ZX group, the traditional Chinese medicine with the same amount of qi, nourishing yin, promoting blood circulation and promoting blood circulation was given in the morning of ZX group, and the same amount of simvastatin was given in the afternoon. In the group of Gavage, N and M, the volume of the corresponding Chinese traditional Chinese medicine was given intragastric administration, once a day, and a total of 1 was given by intragastric administration. 2 weeks. After the end of the stomach, the rats were fasted, and the water for 12h and 2% sodium pentobarbital was injected into the abdominal cavity by 20mg/ kg body weight. The blood was decapitated, and the plasma glucose, total cholesterol, low density lipoprotein cholesterol and high-density lipoprotein cholesterol were detected. Sterol. The thoracic aorta 1-2cm was taken as a specimen in a sterile operation, and was fixed in 4% paraformaldehyde solution. The section line HE staining was made to observe the changes of the vascular endothelial tissue. The enzyme-linked immunosorbent assay was used to detect the serum endothelial lipase (EL). Level. Results: 1 The general situation of the rats in each group was in the course of the experiment, and the rats in the N group were full of spirit and free of activity. The diet, drinking water and urine volume were all normal. The rats in the M group had the symptoms of listlessness, obvious polydipsia, polyphagia, polyuria, gradually reduced body weight, yellow and yellow hair, slow response and the like, and it was in accordance with the clinical

"three or one" of diabetes. The characteristics of s" . The performance of each treatment group in rats in between that normal group and the model group. there was a limb activity in the rat In group M, ZX group. There were 2 deaths and 1 death in the Z group. In group 2, the blood glucose level of the rats was higher than that of the normal group (P <0.05). The blood sugar changes between the group X and the M group were statistically significant (P0.05). There was no significant difference in blood glucose between Z and ZX groups (P <0.05), and the change of blood glucose between Z and ZX groups was not significant (P0.05). No significant, no statistical significance (P0.05). Total cholesterol (TC) and high-density lipoprotein cholesterol (HDL) in each group of rats (1) The level of low-density lipoprotein cholesterol (LDL) was higher than that in the treatment group. Compared with the normal group, the difference was statistically significant (P0.05); the difference between the group Z and the M group was not significant and there was no statistical significance (P0.05); the TC between the X group and the ZX group and the M group was lower and the difference was statistically significant (P0.05); and the difference between the X group and the ZX group was statistically significant (P0.05). There was no significant difference in the TC (P0.05). There was no significant difference in LDL in the M group and in the treatment group (P <0.05), and the difference between the group Z and the M group was not significant (P0.05), and the X group and the ZX group and the M group were not significant (P0.05). There was a significant difference between the group and the group (P <0.05), and the ZX group was the same as that of the group. In group X, LDL decreased and the difference was statistically significant (P0.05). The difference was significant between the treatment group and the control group (P <0.05), and the difference between the treatment group and the M group was significantly higher and the difference was statistically significant (P <0.05). (P0.05); between the three treatment groups There are significant changes in the HDL, and the difference is in series. The results showed that after HE staining, the endothelial cells of group N were clear in texture, normal in shape, uniform in color, flat in the endothelial cells of the thoracic aorta and smooth in the intima. in flat, that middle film is a parallel phase The smooth muscle cells (SMC) and the elastic plate were arranged in the middle membrane of the M group. The inner membrane of the rat thoracic aorta of the M group had the localized thickening and the local uplift and the smooth muscle cells of the inner membrane of the inner membrane were found to be hyperplastic and arranged, and the proliferation of the smooth muscle cells was increased. The elastic plates at the green smooth muscle cells are arranged irregularly, in a wave shape, and even visible elastic plates are broken The changes of the tissue morphology of the thoracic aortic endothelial cells in the treatment group were more than that of the model. The expression of serum endothelial lipase (EL) was detected by enzyme-linked immunosorbent assay (ELISA) in each group, and the levels of EL in all treatment groups were increased. Compared with the normal group, the difference was significant (P <0.05), and the difference between the treatment group and the M group was significantly lower (P <0.05), and the difference was significant (P 0. 0 5) There was a significant difference between the three treatment groups, and the difference was significant (P0. Conclusion: 1. The qi-tonifying and yin-nourishing blood-activating agent can improve the general condition of the diabetic rats, reduce the blood sugar and regulate the lipid metabolism. The qi-tonifying and yin-nourishing blood-activating agent can be used for invigorating qi and nourishing yin and promoting blood circulation. The pathological changes of the vascular endothelial tissue in the thoracic aorta of the diabetic rats have improved, and the effect of the treatment of the diabetic macrovascular endothelial diseases can be provided. The treatment scheme can be provided for clinical treatment.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R259

【引证文献】