[Improvement of the function of islet beta and alpha cells by intervention against glucotoxicity: experiment with rats].
OBJECTIVE
To investigate the effects of intervention against glucotoxicity on improvement of the function and pathological changes of islet beta and alpha cells.


METHODS
Thirty-six male Sprague Dawley rats were randomly divided into four equal groups: normal control (NC) group, fed with standard chow, high-fat (HF) group, fed with extra high-fat chow; diabetes mellitus (DM) control group, fed with high-fat chow for 8 weeks followed by 30 mg/kg streptozotocin injection to establish DM models; and insulin (INS) group, treated with subcutaneous injection of long-acting insulin (glargine, 0.5 U x kg(-1) x d(-1)) for 4 weeks after the establishment of DM models. 48 h, 2 weeks, and 4 weeks after the STZ injection to the 2 DM groups oral glucose tolerance test (OGTT) was performed to all rats. Peripheral blood samples were collected from the caudal vein. Serum insulin level was assayed by radioimmunoassay. Total serum cholesterol (TC) and triglyceride (TG) were measured by enzyme-colorimetric method. By the end of experiment the rats were killed with their pancreases taken out. Immunohistochemistry was used to observe the morphological changes of the islet beta and alpha cells. Beta cell and alpha cell masses were calculated by the proportions of positive area in the islet. Proinsulin mRNA level was detected by RT-PCR. Insulin protein content in islets was detected by Western blotting.


RESULTS
Four weeks after the insulin intervention against glucotoxicity, the fasting blood glucose and blood glucose 2 h after sugar-taking of the INS group were both significantly lower than those of the DM group (both P < 0.01). The relative beta cell mass of the INS group was 0.38 +/- 0.08, significantly bigger, 2.45 times, that of the DM group (0.11 +/- 0.05, P < 0.01). The relative alpha cells mass in islets of the INS group was 0.16 +/- 0.04, significantly lower, by 43%, than that of the DM group (0.28 +/- 0.15, P < 0.01). The insulin contents in beta cells of the INS group was 0.58 +/- 0.03, significantly higher, by 70.6%, than that of the DM group (0.34 +/- 0.14, P < 0.01). The proinsulin mRNA level of the INS group was 1.52 +/- 0.14, significantly higher, by 20.6%, than that of the DM group.


CONCLUSIONS
The morphology of islet beta, alpha cells in diabetic rats was improved by four weeks of Intervention against glucotoxicity improves the pathology of islet beta and alpha cells in diabetic and insulin synthesis.
He
Qing-Hua
QH
Department of Endocrinology, Beijing Hospital, Ministry of Health, Beijing 100730, China.
Zhou
Ying-Sheng
YS
Wang
Zheng
Z
Wang
Shu
S
Mou
Zhong-Qing
ZQ
Li
Jian
J
Chi
Jia-Min
JM
D000818:Animals; D001786:Blood Glucose / Q000378:metabolism; D015153:Blotting, Western; D002784:Cholesterol / Q000097:blood; D003124:Colorimetry / Q000379:methods; D003921:Diabetes Mellitus, Experimental / Q000097:blood / Q000139:chemically induced / Q000503:physiopathology*; D004041:Dietary Fats; D005947:Glucose / Q000633:toxicity*; D015309:Glucose Clamp Technique; D007328:Insulin / Q000097:blood / Q000378:metabolism; D007333:Insulin Resistance; D007515:Islets of Langerhans / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008297:Male; D011384:Proinsulin / Q000235:genetics / Q000378:metabolism; D011863:Radioimmunoassay; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014280:Triglycerides / Q000097:blood
D001786:Blood Glucose; D004041:Dietary Fats; D007328:Insulin; D014280:Triglycerides; D011384:Proinsulin; D002784:Cholesterol; D005947:Glucose