构建糖尿病性肢端坏疽大鼠模型及中药复方的干预效应

背景:糖尿病性肢端坏疽已成为糖尿病致残、致死的重要原因之一。目的:建立糖尿病性肢端坏疽的大鼠模型,观察中药复方对糖尿病性肢端坏疽的干预作用。设计:对比观察实验。单位:河南科技大学临床医学院第一附属医院。材料:选用雄性Wistar大鼠50只,6周龄,体质量(200.0±16.3)g。以普通饲料单笼喂养,室温18～22℃,自由摄食、饮水。随机抽取10只作为空白对照组,其余40只用于造模。方法:实验于2001-10/2002-04在河南科技大学动物房完成。①实验分组:将40只大鼠禁食6h后左下腹腹腔注射链脲佐菌素55.0mg/kg;空白对照组注入同体积的柠檬酸钠缓冲溶液。选用造模成功的20只大鼠,随机分为2组:模型组和治疗组,每组10只。治疗组大鼠于造模成功后每天9:00灌胃中药(60g/kg),共3周;模型组灌胃等体积的生理盐水。第3周末,各组大鼠禁食12h,麻醉后处死取血测血糖、血脂和胰岛素水平;实验过程中记录各组大鼠的每日饮水量。②实验评估:肢端坏疽积分标准:以皮肤颜色发黑、皮肤有轻度开放性病灶、病灶已侵入深部肌肉组织3个等级对糖尿病性肢端坏疽大鼠的四肢进行评分,计算每只大鼠的总积分。对胰岛β细胞分泌的功能进行评价主要观察指标:实验前后大鼠饮水量、体质量、三酰甘油、胆固醇、空腹血糖、血脂和胰岛素水平的变化。结果:纳入Wistar大鼠50只,造模不成功脱落20只,30只进入结果分析。①实验期间,模型组和治疗组饮水量明显高于空白对照组(P<0.01);治疗组随着治疗时间增加饮水量明显下降,模型组随着时间的推移逐渐升高。②治疗后模型组大鼠体质量明显低于治疗前(P<0.01),治疗组有下降趋势,与治疗前比较,差异不明显(P>0.05)。③两组大鼠均有明显肢端坏疽出现(P<0.01)。治疗组大鼠的体质量明显高于模型组(P<0.01),肢端坏疽情况明显好于模型组(P<0.01)。④治疗前治疗组和模型组的空腹血糖水平明显高于空白对照组(P<0.01),而胰岛素水平和胰岛β细胞功能指数明显低于空白对照组(P<0.01)。治疗后两组的空腹血糖水平高于空白对照组(P<0.01),治疗组的明显低于模型组(P<0.01),已接近正常值;两组血脂和胰岛β细胞功能指数均明显低于空白对照组(P<0.01),治疗组明显高于模型组(P<0.01),接近正常值。⑤治疗前其它两组三酰甘油、胆固醇水平明显高于空白对照组(P<0.01)。经过治疗,治疗组三酰甘油、胆固醇明显下降(P<0.01),与空白对照组比较,差异不明显(P>0.05);模型组继续升高,明显高于空白对照组(P<0.01)。结论:血清胰岛素水平升高,血糖、血脂水平下降,可在一定程度上防治糖尿病足的发生、发展。该糖尿病性肢端坏疽模型对药物反应敏感,可用于研究糖尿病足发病机制及药物治疗效果的评价。

Establishing a rat model of diabetic acromelic gangrene and intervention of Chinese compound

BACKGROUND:Diabetic acromelic gangrene(diabetic foot)has become one of the important causes for the disability and death in diabetes mellitus.OBJECTIVE:To establish model of diabetic foot in rat,and observe the interventional effect of Chinese compound on diabetic foot.DESIGN:A comparative observational experiment.SETTING:The First Affiliated Hospital of Clinical Medical College,Henan University of Science and Technology.MATERIALS:Fifty male Wistar rats of 8 weeks old,(200.0±16.3)g,were raised with common feed in separate cage at the room temperature of 18-22℃.and they were free to access of feed and water.Ten rats were randomly selected as the blank control group,and the other 40 were used for model establishment.METHODS:The experiments were carried out in the Animal Room of Henan University of Science and Technology from October 2001 to April 2002.①Grouping:The 40 rats were fasted for 6 hours,and then treated with intraperitoneal injection of streptozotocin(55.0 mg/kg),while the 10 rats in the blank control group were injected with isovolume sodium citrate buffer solution.20 models were successfully established,and they were randomly divided into model group(n=10)and treatment group(n=10).Rats in the treatment group were treated for 3 weeks with intragastric perfusion of Chinese compound(60 g/kg)at 9:00 every day after model establishment,and those in the model group were given intragastric perfusion of isovolume saline.At the end of the third week,the rats were all killed under anesthesia after fasted for 12 hours,blood samples were collected to determine the levels of fasting blood glucose,blood lipids and insulin.The daily amount of drinking water was recorded in each group during the experiment.②Scoring standards for acromelic gangrenes:The limbs rats with diabetic foot were scored by three grades,including dark skin,mild open focus on skin,and focus had invaded deep muscular tissue.The total score of each rat was calculated.The beta-cell function index (HBCI)was also evaluated.MAIN OUTCOME MEASURES:The changes of the amount of drinking water,body mass and levels of triglyceride,cholesterol,fasting blood glucose,blood lipids and insulin were observed before and after treatment.RESULTS:Totally 50 Wistar rats were used.20 of them were excluded due to unsuccessful model establishment,and the other 30 rats were involved in the final analysis of results.①The amount of drinking water was obviously higher in the model group and treatment group than in the blank control group during the experiment(P＜0.01).As the treatmentlasted,the amount of drinking water was obviously decreased in the treatment group,but gradually increased in the model group.②After treatment,the body mass was obviously lower than that before treatment in the model group(P＜0.01).but had a descending trend without obvious difference as compared with that before treatment in the treatment group(P＞0.05).③Obvious acromelic gangrenes were obvious in both groups(P＜0.01).The body mass in the treatment group was obviously higher than that in the model group(P＜0.01),and the conditions of acromelic gangrene were obviously better than those in the model group(P＜0.01).④Before treatment,the levels of fasting blood glucose in the treatment group and model group were obviously higher than that in the blank control group(P＜0.01),while the levels of insulin and HBCl were obviously lower than those in the blank control group(P＜0.01).After treatment,the levels of fasting blood glucose in the treatment group and model group were obviously higher than that in the blank control group(P＜0.01),and it was obviously lower in the treatment group than in the model group(P＜0.01),it was close to the normal value in the treatment group.⑤The levels of triglyceride and cholesterol before treatment were obviously higher in the treatment group and model group than in the blank control group(P＜0.01).After treatment,the levels of triglyceride and cholesterol in the treatment group were obviously decreased(P＜0.01), which were not obviously different from those in the blank control group (P＞0.05), while those in the model group were increased continuously,and obviously higher than those in the blank control group(P＜0.01).CONCLUSION:Increasing the serum level of insulin and decreasing the levels of blood glucose and blood lipids can prevent and treat the occurrence and development of diabetic foot to some extent.This model of diabetic foot is sensitive to drug,and can be used to investigate the pathogenesis of diabetic foot and evaluate the effect of drug therapy.