Changes of macrovascular endothelial ultrastructure and gene expression of endothelial nitric oxide synthase in diabetic rats.
BACKGROUND
The most intimidatory pathological changes in patients with DM are cardiovascular illnesses, which are the major causes of death in diabetic patients and are far more prevalent than in nondiabetics because of accelerated atherosclerosis. In this study, we tried to clarify the changes in macrovascular endothelial ultrastructure and in the gene expression of endothelial nitric oxide synthase (eNOS)mRNA in diabetic rats.


METHODS
The study was conducted on 52 of 10-week old Sprague Dawley (SD) rats with body weight of (320 +/- 42) g. SD rats were divided into: experimental group treated with a single intraperitoneal injection of streptozotocin (STZ, 60 mg/kg), (male, n = 20, diabetes mellitus (DMM)); female, n = 12, diabetes mellitus female (DMF)) and control group (male, n = 10, diabetes mellitus male control (DMMC); female, n = 10, diabetes mellitus female control (DMFC)). Four weeks after treatment, half of the rats were sacrificed; the remainders were sacrificed ten weeks after treatment. One part of the abdominal aortic sample was stored under glutaraldehyde (volume fraction psiB = 2.5%). After the process of chemical fixation, chemical dehydration, drying and conductivity enhancement, all samples were observed and photographed using scanning electron microscopy (Leica-Stereoscan 260, England). The other part of the abdominal aortic sample was treated with liquid nitrogen and the expression of eNOSmRNA was assessed by semi-quantitative RT-PCR.


RESULTS
The aortic lumen of both experimental groups adsorbed much more debris than that of either control group. The endothelial surfaces of diabetic rats were coarse, wrinkled and protuberant like fingers or villi. The vascular endothelial lesions of diabetic male rats were very distinct after 4 weeks, and as obvious as those at 10 weeks. The vascular endothelial lesions of diabetic female rats were not severe at 4 weeks and only became marked after 10 weeks. In both males and females, the abdominal aortic eNOSmRNA content of 4 weeks and 10 weeks diabetic rats was very significantly lower (P < 0.01) than that of controls.


CONCLUSIONS
Aortic endothelial ultrastructure in DM rats is injured compared with controls. Abnormal changes of aortic endothelia in male DM rats are more obvious than those in females. Expression of abdominal aortic eNOSmRNA content of DM rats is significantly lower than that of controls.
Lu
Ying-li
YL
Department of Endocrinology, Sir Run Run Shaw Hospital, Hangzhou, China.
Hu
Shen-jiang
SJ
Shen
Zhou-jun
ZJ
Shao
Yi-chuan
YC
D000818:Animals; D001012:Aorta, Abdominal / Q000201:enzymology; D001786:Blood Glucose / Q000032:analysis; D003921:Diabetes Mellitus, Experimental / Q000201:enzymology / Q000473:pathology*; D004730:Endothelium, Vascular / Q000648:ultrastructure*; D005260:Female; D008297:Male; D019001:Nitric Oxide Synthase / Q000235:genetics*; D052250:Nitric Oxide Synthase Type III; D012333:RNA, Messenger / Q000097:blood; D051381:Rats; D017207:Rats, Sprague-Dawley; D012737:Sex Factors; D013311:Streptozocin
D001786:Blood Glucose; D012333:RNA, Messenger; D013311:Streptozocin; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; C496316:Nos3 protein, rat