摘要
目的:观察参附注射液对糖尿病大鼠心肌超微结构的影响并探索其可能机制。方法:实验于2003-12/2005-01在重庆医科大学生理教研室完成。①实验分组:30只Wistar大鼠随机分为正常对照组10只、糖尿病组20只。②实验方法:所有大鼠禁食12h后,糖尿病组大鼠以链脲佐菌素(以0.1mol/L,pH=4.2的枸橼酸缓冲液溶解)按60mg/kg经腹腔注射,正常对照组以等量的枸橼酸缓冲液腹腔注射。72h后,大鼠剪尾取静脉血测定空腹血糖,当空腹血糖>16.67mmol/L并有多饮、多食、多尿者,即为1型糖尿病造模成功。糖尿病组大鼠又按分层随机法分为单纯糖尿病组和参附注射液处理组,每组10只,参附注射液组以参附注射液10mL/kg腹腔注射,1次/d。正常对照组和单纯糖尿病组大鼠均以生理盐水10mL/kg腹腔注射。所有大鼠均以普通食物喂养,自由饮水和单笼饲养20周。③实验评估:20周后测定各组血糖、血脂、血清胰岛素、血清与心肌超氧化物歧化酶活性和丙二醛含量;免疫组化法检测心肌细胞膜葡萄糖转运蛋白4mRNA蛋白表达;观察各组大鼠胰岛细胞的病理结构以及心肌超微结构的变化。结果:30只大鼠均进入结果分析。①空腹血糖、血清胰岛素水平:造模后20周,参附注射液组的空腹血糖明显低于造模3d时水平(P<0.05),明显低于单纯糖尿病组,但高于正常对照组(P<0.01);单纯糖尿病组空腹血糖明显高于造模3d时水平(P<0.01)。造模后20周,参附注射液组血清胰岛素水平明显低于正常对照组,但显著高于单纯糖尿病组(P<0.05)。②血脂水平:参附注射液组和单纯糖尿病组的总三酰甘油、总胆固醇和低密度脂蛋白胆固醇水平明显高于正常对照组(P<0.05),参附注射液组各值明显低于单纯糖尿病组(P<0.05);单纯糖尿病组和参附注射液组高密度脂蛋白胆固醇水平明显低于正常对照组(P<0.05),参附注射液组高密度脂蛋白胆固醇水平明显高于单纯糖尿病组(P<0.05)。③血清和心肌超氧化物歧化酶活性及丙二醛含量:超氧化物歧化酶活性:参附注射液组和单纯糖尿病组明显低于正常对照组(P<0.01),参附注射液组明显高于单纯糖尿病组(P<0.05);血清丙二醛的含量:参附注射液组和单纯糖尿病组明显高于正常对照组(P<0.05),参附注射液组明显低于单纯糖尿病组(P<0.05)。④葡萄糖转运蛋白4mRNA阳性表达:参附注射液组吸光度显著低于正常对照组(P<0.01),明显高于单纯糖尿病组(P<0.01),而参附注射液组的面密度值显著低于正常对照组(P<0.01),明显高于单纯糖尿病组(P<0.01)。⑤光镜观察:参附注射液组大鼠胰岛细胞水肿或空泡样变比单纯糖尿病组数量少而且更轻;透视电镜观察参附注射液组其心肌细胞线粒体和心肌毛细血管内皮细胞的水肿较单纯糖尿病组程度更轻。结论:参附注射液对糖尿病大鼠心肌超微结构具有保护作用,这与其清除氧自由基和抗脂质过氧化物有关;参附注射液对糖尿病大鼠健存胰岛细胞的保护,提高了糖尿病大鼠的血清胰岛素水平,降低了血糖、血脂,对糖尿病大鼠心肌内微血管内皮细胞和心肌细胞线粒体的保护具有促进作用。
AIM： To investigate the effect of Shenfu injection （SFI） on the ultrastructure of myocardium in diabetes mellitus （DM） rats, and explore the possible mechanism. METHODS： The experiments were carded out in the Department of Physiology at Chongqing Medical University from December 2003 to January 2005.①Thirty Wisatr rats were randomly divided into normal control group （n =10） and DM group （n =20）.②All the rats were fasted for 12 hours, then those of DM group were treated with 60 mg/kg straptozotocin （dissolved in 0.1 mol/L citric acid buffer, pH=4.2） by intraperitoneal injection, while those of normal control group were injected with equal volume of citric acid buffer solution. And 72 hours later, venous blood was sampled from rat tails to detect fasting blood glucose （FBG）. Those rats appearing polydipsia, polyphagia and polyuria, whose FBG were more than 16.7 mmol/L indicated successful animal model of type 1 DM According to stratified random, DM group was assigned to the diabetic control group （C-DM group） and the SFI treated group （S-DM group）, 10 rats in each. The rats in S-DM group were injected with SFI （10 mL/kg） into peritoneal cavity once a day. The rats in control group and C-DM group were injected with 10 mL/kg saline by peritoneal cavity. All the rats were fed routinely for 20 weeks.③At the end of week 20, the FBG, blood fat, serum insulin, superoxide dismutase （SOD） and malondialdehyde （MDA） in serum and myocardium were assayed. The expressions of glucose transport protein 4 mRNA on the myocardial cytomembrane were detected by immunohistochemical assay; the pathologic changes of islets of pancreas and myocardial ultrastructure of rats in three groups were also observed. RESULTS： Thirty rats were all involved in the result analysis. ①Levels of FBG and serum insulin： The FBG of rats in S-DM group was remarkably lower than that of C-DM group but much higher than that of control group （P 〈 0.01） at the end of 20 weeks. FBG of rats at the end of 3 weeks was obviously higher than that at the end of 20 weeks in S-DM group （P 〈 0.05） but obviously lower in C-DM group （P 〈 0.01 ）. The serum insulin of rats in S-DM group was obviously higher than that of C-DM group but significantly lower than that of control group （P 〈 0.05）.②Blood fat level： The levels of total triglycedde, cholesterol, and low-density lipoprotein cholesterol were obviously higher in S-DM group and C-DM group than those of control group （P 〈 0.05）, and all obviously lower in S-DM group than those of C-DM group （P 〈 0.05）; The levels of high-density lipoprotein cholesterol in C-DM group and S-DM group were obviously lower than that of control group （P 〈 0.05）, and the level was significantly higher in S-DM group than in C-DM group （P 〈 0.05）.③Contents of SOD and MDA in serum and myocardium： SOD in S-DM group was much more than that of C-DM group （P〈 0.05）, but much less than that of control group （P 〈 0.01） while MDA was much less than that of C-DM group （P 〈 0.05）, but much more than that of control group （P 〈 0.05）. ④Expressions of glucose transport protein 4 mRNA on cardiac cytomembrane of rats in S-DM group were significantly less than those of control group （P 〈 0.01 ） while obviously more than those of C-DM group （P 〈 0.01 ）. ⑤Under optic microscope, there was much less cellular edema or vacuole of pancreatic islets in rats from S-MD group compared with that of C-DM group. Under transmission electron microscope, the edema on myocardial mitochondria and endothelial cells of cardiac capillary in C-DM group was much more serious than that of S-DM group. CONCLUSION： SFI can protect the ultrastructure of myocardium in DM rats and remove oxygen-derived free radicals and resist lipid oxidation, moreover, it has the protective effect on healthy islets of pancreas in diabetic rats, which does facilitate the protection on endothelial cells of cardiac capillary and the mitochondria of myocardium, because of the increase in serum insulin and decrease in blood glucose and fat.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2007年第29期5712-5716,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
