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PROINSULIN C-PEPTIDE CAN INHIBIT THE EARLY CHANGES OF GLOMERULOPATHY IN DIABETIC MICE
Poster Abstracts / Cardiovascular Pathology 13 (2004) S80–S138 phospho-tyrosine immunoreactivity and hyperphosphorylated beta-catenin. Vascular endothelial growth factor levels were higher in these placentae. A 4-fold increase in endothelial proliferation was seen coupled with a greater total length of capillaries without any change in luminal diameter. Molecular perturbations of vascular endothelial cadherin and beta-catenin occur in fetoplacental vessels from pregnancies complicated by Type 1 diabetes. Phosphorylation and loss of these molecules from adherens junctional domains may be influenced in part by the elevated levels of placental vascular endothelial growth factor. Perturbations of the junctional proteins may explain the observed breach in barrier integrity and may contribute to mechanisms driving proliferation and increases in capillary length. The Wellcome Trust
P222 THE SUPPLEMENTARY EFFECTS OF HIGH DIETARY FIBER RICE (SUWON 464) ON BLOOD PRESSURE AND LIPID METABOLISM IN SPONTANEOUSLY HYPERTENSIVE RATS. Sunghyeon Lee, Hongju Park, Soyoung Cho, Hyunjin Jeong, Hunggu Hwang, Yeonsook Lee. National Rural Living Science Institute, NIAST, RDA, Suwon, Korea, National Crop Experiment Station, RDA, Suwon, Korea, Seoul National University, Seoul, Korea. Objectives: The high dietary fiber rice (Suwon 464) was developed by Rural Development Administration in Korea to increase dietary fiber content in rice as the source of Korean main dish. This study was carried out to investigate the supplementary effects of this high dietary fiber rice on blood pressure and lipid metabolism in spontaneously hypertensive rats. Materials and methods: After analyzing the nutritional contents of polished and brown rice of Suwon 464 and Ilpum, mother of Suwon 464, we supplied 5 kinds of experimental diets including cornstarch diet as a control (CO), polished (IP) and brown rice (IB) of Ilpum, and polished (SP) and brown rice (SB) of Suwon 464, to 5 groups of spontaneously hypertensive rats for 8 weeks respectively. Diet intake, body weight, systolic and diastolic blood pressure, and lipids levels of serum, liver and feces were measured. Results: The high dietary fiber rice (Suwon 464) contained dietary fiber two times as much as normal rice (Ilpum) in both polished and brown rice. When five groups of spontaneously hypertensive rats were fed one of five kinds of diets (CO, IP, IB, SP, SB) for 8 weeks, body weight gain was lower in four rice intake groups though there was no significant difference in diet intake among experimental groups. The systolic and diastolic blood pressure levels were decreased 18% and 16% in SP group compared with CO group, respectively. LDL-cholesterol level was significantly lower in SB group and HDL-cholesterol level was significantly higher in IB, SP and SB groups though serum triglyceride and total cholesterol levels have no significant difference among experimental groups. The hepatic total lipid, triglyceride and total cholesterol concentrations were significantly lower in four groups fed rice diets, and these three levels were significantly lower in SB group than in IP group. Total lipid, triglyceride and total cholesterol contents excreted to feces were significantly higher in IB, SP and SB groups, and these three levels were significantly higher in SB group than in IP and IB groups. Conclusion: These results suggest that the high dietary fiber rice (Suwon 464) can decrease blood pressure, serum LDL-cholestrol, hepatic total lipid, triglyceride and total cholesterol by increasing fecal lipid, and increase serum HDL-cholesterol. We expect that high dietary fiber rice (Suwon 464) can control blood pressure, and serum and hepatic lipid levels.
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P223 PROINSULIN C-PEPTIDE CAN INHIBIT THE EARLY CHANGES OF GLOMERULOPATHY IN DIABETIC MICE. Yoshiro Maezawa, Koutaro Yokote, Kiriko Sonezaki, Masaki Fujimoto, Minoru Takemoto, Harukiyo Kawamura, Kazuki Kobayashi, Takahiko Tokuyama, Aki Takada, Seijiro Mori, John Wahren, Yasushi Saito. The Second Dept of Internal Medicine Chiba University, Inohana, Japan, Sect of Clinical Physiology, Karolinska Institute, Stockholm, Sweden. Diabetes mellitus is an important risk factor of atherosclerosis. Diabetic nephropathy is also closely related to atherosclerosis, and recent studies have revealed that microalbuminuria is an independent risk factor of cardiovascular complications. Although a number of mechanisms have been proposed for the pathogenesis of diabetic nephropathy, its precise molecular basis remains unclear. Prompted by recent reports that C-peptide may inhibit the progression of nephropathy in type 1 diabetes, we have studied the renal effects of C-peptide by analyzing the glomerular gene expression in streptozotocin-induced (100 mg/kg BW/24 h for 3 days) diabetic mice (C57Bl/6). Fourteen days after induction of diabetes the animals received rat C-peptide intraperitoneally (i.p.) as 6 injections of 70 nmol/kg BW each over 24 h. Diabetic control mice were given saline i.p. Mice urine was collected by use of metabolic cages, and urinary albumin excretion was measured by ELISA. The animals were then sacrificed and glomeruli were isolated with the magnet microbeads method. RNA was extracted from the glomeruli and realtime PCR (rtPCR) was used to quantify the expression of genes associated with diabetic glomerulopathy (TGF-beta, type IV collagen). The results reveal that C-peptide decreases urinary albumin excretion from 0.75 ± 0.19 to 0.37 ± 0.20 mg/min ( 44% ( P < .05)). Nephrin, the podocyte foot process slit membrane protein, was excreted into urine in diabetic mice (1.0 ± 0.38, arbitrary unit, Western blot analysis), whereas it was not detectable in urine of C-peptide treated mice (0.08 ± 0.04, arbitrary unit), suggesting that C-peptide inhibits the diabetesinduced loss of nephrin into urine. Expression of TGF-beta in glomeruli was up-regulated 1.6 fold in diabetic mice, but it was not significantly changed by C-peptide treatment. Glomerular gene expression of type IV collagen (alpha3) was up-regulated 2.2 fold in diabetic mice, and this elevation was inhibited by 22% in C-peptide treated mice ( P < .05). In additional studies using a temperature sensitive SV40-induced mice podocyte cell line Western blot analysis and rtRCR showed that C-peptide (0.1, 0.5 and 1 nmol/l) inhibits TGF-beta induced up-regulation of type IV collagen by 60% ( P < .05), suggesting that C-peptide suppresses TGF-beta mediated intracellular signaling. The present results extend previous findings and add a new perspective on the molecular effects of C peptide in diabetic nephropathy. It is concluded that short-term C-peptide administration to diabetic mice significantly prevents several of the early changes of diabetic glomerulopathy.
P224 IS FIBRINOGEN AND D-DIMERS DETERMINATION USEFUL FOR PROGNOSIS OF DIABETIC POLYVASCULAR DISEASE. Bosevski Marijan, Kostoska Stojanka, Tosev Slavco, Meskoska Snezana, Borozanov Vladimir, Spiroska Vera, Peovska Irena. University Institute for Cardiovascular Diseases, Faculty of Medicine, Skopje, Macedonia, Department of Hemostasis and Thrombosis, Clinical Center, University Institute for Cardiovascular Diseases, Faculty of Medicine. Subject: Diabetes mellitus has been recognized as hypercoagulable state. We made hypothesis that hemostatic parameters could predict prognosis of diabetic vascular disease. Patients and methods: 82 patients with Insulin Dependent Diabetes mellitus and coexistent peripheral arterial disease and coronary artery disease were included in 36 months study (from March 1st 2000 to March 1st 2003). 62 men and 20 women with age 63,6+6,9 were evaluated for the following parameters: current smoking, ankle-brachial index, claudicating
P222 THE SUPPLEMENTARY EFFECTS OF HIGH DIETARY FIBER RICE (SUWON 464) ON BLOOD PRESSURE AND LIPID METABOLISM IN SPONTANEOUSLY HYPERTENSIVE RATS. Sunghyeon Lee, Hongju Park, Soyoung Cho, Hyunjin Jeong, Hunggu Hwang, Yeonsook Lee. National Rural Living Science Institute, NIAST, RDA, Suwon, Korea, National Crop Experiment Station, RDA, Suwon, Korea, Seoul National University, Seoul, Korea. Objectives: The high dietary fiber rice (Suwon 464) was developed by Rural Development Administration in Korea to increase dietary fiber content in rice as the source of Korean main dish. This study was carried out to investigate the supplementary effects of this high dietary fiber rice on blood pressure and lipid metabolism in spontaneously hypertensive rats. Materials and methods: After analyzing the nutritional contents of polished and brown rice of Suwon 464 and Ilpum, mother of Suwon 464, we supplied 5 kinds of experimental diets including cornstarch diet as a control (CO), polished (IP) and brown rice (IB) of Ilpum, and polished (SP) and brown rice (SB) of Suwon 464, to 5 groups of spontaneously hypertensive rats for 8 weeks respectively. Diet intake, body weight, systolic and diastolic blood pressure, and lipids levels of serum, liver and feces were measured. Results: The high dietary fiber rice (Suwon 464) contained dietary fiber two times as much as normal rice (Ilpum) in both polished and brown rice. When five groups of spontaneously hypertensive rats were fed one of five kinds of diets (CO, IP, IB, SP, SB) for 8 weeks, body weight gain was lower in four rice intake groups though there was no significant difference in diet intake among experimental groups. The systolic and diastolic blood pressure levels were decreased 18% and 16% in SP group compared with CO group, respectively. LDL-cholesterol level was significantly lower in SB group and HDL-cholesterol level was significantly higher in IB, SP and SB groups though serum triglyceride and total cholesterol levels have no significant difference among experimental groups. The hepatic total lipid, triglyceride and total cholesterol concentrations were significantly lower in four groups fed rice diets, and these three levels were significantly lower in SB group than in IP group. Total lipid, triglyceride and total cholesterol contents excreted to feces were significantly higher in IB, SP and SB groups, and these three levels were significantly higher in SB group than in IP and IB groups. Conclusion: These results suggest that the high dietary fiber rice (Suwon 464) can decrease blood pressure, serum LDL-cholestrol, hepatic total lipid, triglyceride and total cholesterol by increasing fecal lipid, and increase serum HDL-cholesterol. We expect that high dietary fiber rice (Suwon 464) can control blood pressure, and serum and hepatic lipid levels.
S87
P223 PROINSULIN C-PEPTIDE CAN INHIBIT THE EARLY CHANGES OF GLOMERULOPATHY IN DIABETIC MICE. Yoshiro Maezawa, Koutaro Yokote, Kiriko Sonezaki, Masaki Fujimoto, Minoru Takemoto, Harukiyo Kawamura, Kazuki Kobayashi, Takahiko Tokuyama, Aki Takada, Seijiro Mori, John Wahren, Yasushi Saito. The Second Dept of Internal Medicine Chiba University, Inohana, Japan, Sect of Clinical Physiology, Karolinska Institute, Stockholm, Sweden. Diabetes mellitus is an important risk factor of atherosclerosis. Diabetic nephropathy is also closely related to atherosclerosis, and recent studies have revealed that microalbuminuria is an independent risk factor of cardiovascular complications. Although a number of mechanisms have been proposed for the pathogenesis of diabetic nephropathy, its precise molecular basis remains unclear. Prompted by recent reports that C-peptide may inhibit the progression of nephropathy in type 1 diabetes, we have studied the renal effects of C-peptide by analyzing the glomerular gene expression in streptozotocin-induced (100 mg/kg BW/24 h for 3 days) diabetic mice (C57Bl/6). Fourteen days after induction of diabetes the animals received rat C-peptide intraperitoneally (i.p.) as 6 injections of 70 nmol/kg BW each over 24 h. Diabetic control mice were given saline i.p. Mice urine was collected by use of metabolic cages, and urinary albumin excretion was measured by ELISA. The animals were then sacrificed and glomeruli were isolated with the magnet microbeads method. RNA was extracted from the glomeruli and realtime PCR (rtPCR) was used to quantify the expression of genes associated with diabetic glomerulopathy (TGF-beta, type IV collagen). The results reveal that C-peptide decreases urinary albumin excretion from 0.75 ± 0.19 to 0.37 ± 0.20 mg/min ( 44% ( P < .05)). Nephrin, the podocyte foot process slit membrane protein, was excreted into urine in diabetic mice (1.0 ± 0.38, arbitrary unit, Western blot analysis), whereas it was not detectable in urine of C-peptide treated mice (0.08 ± 0.04, arbitrary unit), suggesting that C-peptide inhibits the diabetesinduced loss of nephrin into urine. Expression of TGF-beta in glomeruli was up-regulated 1.6 fold in diabetic mice, but it was not significantly changed by C-peptide treatment. Glomerular gene expression of type IV collagen (alpha3) was up-regulated 2.2 fold in diabetic mice, and this elevation was inhibited by 22% in C-peptide treated mice ( P < .05). In additional studies using a temperature sensitive SV40-induced mice podocyte cell line Western blot analysis and rtRCR showed that C-peptide (0.1, 0.5 and 1 nmol/l) inhibits TGF-beta induced up-regulation of type IV collagen by 60% ( P < .05), suggesting that C-peptide suppresses TGF-beta mediated intracellular signaling. The present results extend previous findings and add a new perspective on the molecular effects of C peptide in diabetic nephropathy. It is concluded that short-term C-peptide administration to diabetic mice significantly prevents several of the early changes of diabetic glomerulopathy.
P224 IS FIBRINOGEN AND D-DIMERS DETERMINATION USEFUL FOR PROGNOSIS OF DIABETIC POLYVASCULAR DISEASE. Bosevski Marijan, Kostoska Stojanka, Tosev Slavco, Meskoska Snezana, Borozanov Vladimir, Spiroska Vera, Peovska Irena. University Institute for Cardiovascular Diseases, Faculty of Medicine, Skopje, Macedonia, Department of Hemostasis and Thrombosis, Clinical Center, University Institute for Cardiovascular Diseases, Faculty of Medicine. Subject: Diabetes mellitus has been recognized as hypercoagulable state. We made hypothesis that hemostatic parameters could predict prognosis of diabetic vascular disease. Patients and methods: 82 patients with Insulin Dependent Diabetes mellitus and coexistent peripheral arterial disease and coronary artery disease were included in 36 months study (from March 1st 2000 to March 1st 2003). 62 men and 20 women with age 63,6+6,9 were evaluated for the following parameters: current smoking, ankle-brachial index, claudicating