potassium, calcium, phosphorous, bilirubin, ALT, AST, albumin, complete lipid profile, hs-CRP and Orsomucoid 2 (ORM2) assay. Electrocardiogram (ECG), Echocardiography and B-mode ultrasonography of left and right carotid arteries were also assessed. RESULTS: Serum orsomucoid 2 and hs-crp were significantly higher in group 1 compared to group 2 at (p0.001). Serum orosomucoid 2 at cut off 11.05 ng/L could discriminate patients with and without subclinical atherosclerosis. There was Positive correlation between serum orsomucoid level, hs-CRP and each of CIMT, LDL-C, and TG while negative correlation was found with HDL-C. Mean CIMT presented positive correlation with total cholesterol level, triglycerides and low-density lipoprotein. CONCLUSIONS: Our findings suggest that orsomucoid 2 and hs-CRP could be used effectively as biomarkers for subclinical atherosclerosis in lean T2DM patients. SP449 ROLE OF ACTIVATING TRANSCRIPTION FACTOR 3 (ATF3) IN DIABETIC NEPHROPATHY Jeong Lim 1 , Jung Joa 1 , Ji Yoo 1 , Jung Ghee 1 , Jin Cha 1 , Hye Min 2 , Ji Lee 2 , Young Kang 1 , Jee Han 3 , Dae Cha 1 1 Korea University Ansan Hospital, Ansan-si, Republic of Korea, 2 Wonkwang University Gunpo Hospital, Ansan-si, Republic of Korea and 3 Inha University Hospital, Incheon, Republic of Korea INTRODUCTION: Activating Transcription Factor 3 (AFT3) is a transcriptional factor in response to stress, and reported that its expression is activated by ER stress and oxidative stress. Recent reports suggest that ATF3 involves in the development of diabetes through modulation of glucose regulatory molecules such as GCK. Therefore, persistent ATF3 expression induced by excessive ROS or ER stress likely has detrimental effects, overwhelming its initial compensatory role. Nonetheless, the exact functional role of ATF3 as a target molecule responsible for oxidative stress-mediated diabetic kidney injury is largely unknown. Hence, the aim of this study was to investigate the role of ATF3 in diabetic nephropathy METHODS: To investigate the chronological changes of ATF3 in kidney in accordance with the progression of diabetic nephropathy, 6 week old db/m and db/db mice were sacrificed at 8, 12, 16 and 20weeks. Plasma and urinary levels of ATF3 protein levels were measured as well as the changes in renal morphological and functional changes. In addition, we examined the effects of high glucose and angiotensin-II on the synthesis of ATF3 in cultured podocytes and PTCs. RESULTS: Although plasma ATF3 concentrations did not show significant differences between diabetic and control mice, Urinary levels of ATF3 were significantly higher in the diabetic mice than in the controls. Interestingly, urinary excretion of ATF3 persistently increased during the course of diabetic nephropathy in diabetic mice. In renal tissues, ATF3 expression was significantly increased in the diabetic kidney even in the early stage of diabetic nephropathy. Interestingly, increased ATF3 expression was associated with activation of NOX1, NOX4 and Smad 2/3 and NF-kB. In cultured podocytes and PTCs, high glucose, angiotensin II stimuli markedly increased ATF3 synthesis and secretion. CONCLUSIONS: These findings suggest that ATF3 is activated in early stage of diabetic environment, may be an important pathogenic role in diabetic nephropathy. SP450 SGLT2 INHIBITOR PROTECTED RENAL PROXIMAL TUBULES FROM GLUCOSE-INDUCED APOPTOSIS VIA BCL-2-CASPASE PATHWAY Wen-Chin Lee 1 , You-Ying Chau 2 , Chiu-Hua Chen 1 , Jin-Bor Chen 1 1 Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, Taiwan and 2 Queen’s Medical Research Institute, The University of Edinburgh, Edinburgh, United Kingdom INTRODUCTION: Sodium-glucose cotransporter 2 (SGLT2) inhibitors have been demonstrated to retard progression of kidney disease in diabetic patients. Reported mechanisms for this renal benefit include tubuloglomerular feedback, reducing body weight and lowering blood pressure. However, little is known about the direct kidney mechanism occurred in the exact SGLT2 inhibitors’ action site. In this study, we aimed to examine the protection effects of SGLT2 inhibitors in glucose-treated renal proximal tubular cells. METHODS: Human proximal tubular cells (PTC), HK2 cells, were used as a cellular model to test the central hypothesis of this study. Cells were treated and grouped as normal glucose (NG) (5mM), high glucose (HG) (30mM), HG+empagliflozin 100nM, and HG+ empagliflozin 500nM. TUNEL assay was used to demonstrate glucose induced cellular apoptosis and to examine the rescue effects of empagliflozin on it. Western blots were used to investigate the changes in the expression of apoptosis- related proteins. RESULTS: Compared with NG, TUNEL assay showed that HG markedly increased cellular apoptosis. The increased cellular apoptosis was rescued by empagliflozin in a dose-dependent manner. Western blots showed HG caused down-regulation of Bcl-2 (40%) and up-regulation of t-Bid/ Bid (70%), Bax (126%) and cytochrome C (52%). HG also triggered the activation of caspase-8, 9, 3. Therapeutic dose of empagliflozin (500nM) was shown to reverse the alterations in the expression of these proteins. CONCLUSIONS: Our results suggested that SGLT2 inhibitor protected renal PTC from apoptosis by regulating Bcl-2-Caspase pathway. This could be one of the novel mechanisms explaining the renal benefits demonstrated in the landmark clinical trial. SP451 THE THERAPEUTIC EFFICACY OF WATER SOLUBLE COENZYME Q10 IN EXPERIMENTAL MODEL OF TACROLIMUS INDUCED DIABETES MELLITUS Quan Yi 1 , Luo Kang 2 , Cui Sheng 1 , Shin Yoo Jin 1 , Lim Sun Woo 1 , Yang Chul Woo 3 1 The Catholic University of Korea, Seoul, Republic of Korea, 2 The saint maria hospital of catholic university, Seoul, Republic of Korea and 3 Seoul St. Mary’s Hospital, The Catholic University of Korea, Seoul, Republic of Korea INTRODUCTION: Coenzyme Q10 has a central role in the generation of cellular bioenergy and its regulation, but its use is limited due to poor absorption profiles. We have developed a natural product based micelle formulation of CoQ10 (CoQ10-M), its effect in experimental model of tacrolimus (TAC)-induced diabetes mellitus (DM). METHODS: Rats were daily treated with vehicle (VH), TAC(1.5mg/kg, subcutaneous), TAC+CoQ10 (20mg/kg, P.O.) or TAC+CoQ10-M (20mg/kg, P.O.) under 0.01% salt diet for 4weeks. The control of hyperglycemia by CoQ10-M was evaluated by intraperitoneal glucose tolerance test (IPGTT), and the protective effect of CoQ10-M on pancreatic islet was evaluated with oxidative stress, islet size, apoptosis, and mitochondria morphology. RESULTS: IPGTT revealed that the TAC+CoQ10 group failed to control TAC induced hyperglycemia but the TAC+CoQ10-M group significantly decreased blood sugar level at fasting and each time point compared with the TAC group. The TAC group significantly increased the serum 8-OHdG, a marker of oxidative stress, compared with the VH group (VH: 0.8 6 0.1 vs. TAC: 1.3 6 0.1; P<0.05). The addition of CoQ10 did not but CoQ10-M significantly reduced the 8-OHdG expression (TAC+CoQ10-M: 1.0 6 0.04 vs. TAC group; P<0.05). Thus, CoQ10 did not but CoQ10-M increases the decrescent islet size induced by TAC. In apoptotic cell death, the TAC+CoQ10 group did not but the TAC+CoQ10-M group markedly decreased the number of TUNEL- positive cells induced by TAC. At the subcellular level, The TAC group showed markedly decreased the number and size of mitochondria compared with the VH group. Treatment of CoQ10 did not but CoQ10-M restored those parameters compared with the TAC group. CONCLUSIONS: Conclusions: Water soluble CoQ10 has excellent therapeutic efficacy compared with conventional CoQ10 in TAC-induced DM, and it may provide a new strategy to prevent TAC-induced pancreatic injury. SP452 RENAL MMP-10, IS INCREASED IN EARLY DKD IN DB/DB MODEL AND ANGIOTENSIN RECEPTOR BLOCKADE NORMALIZED ITS EXPRESSION Jose ´ Mar  ıa Mora-Gutie ´ rrez 1 , Maria Jose Soler Romeo 2 , Mar  ıa Fernandez-Seara 1 , Omar Gonzalez Arostegui 1 , Marta Riera 3 , Jose Antonio Paramo 1 , Nuria Garcia- Fernandez 1 1 Clinica Universidad de Navarra, Pamplona, Spain, 2 Hospital Universitari del Vall d  Hebron, Barcelona, Spain and 3 Institut Hospital del Mar d’Investigacions Me `diques IMIM, Barcelona, Spain INTRODUCTION: Matrix metalloproteinases (MMPs) play a role in various pathogenic mechanisms involved in microvascular complications of diabetes mellitus (DM). Diabetic mice, knock-out for MMP-10, showed minor morphologic kidney alterations and better renal function compared to wild-types. Recent data demonstrated increased renal expression of MMP-10 in early primary glomerular disease. Renin-angiotensin (RAS) blockage inhibits MMP-2 activation in diabetic rats. Additionally, MMP-9 expression, triggered by advanced glycation end products, was attenuated by Olmesartan, but no data has been reported linking RAS and renal MMP- 10 expression. The objective is to study renal MMP-10 expression in a mouse model of type-2 DKD, and its potential modulation by RAS blockade. METHODS: Five-week-old male db/db and db/m were divided into four groups (n=6): db/db; db/db Telmisartan-treated (5mg/kg/day, starting at 8 weeks of age); db/m; db/m Telmisartan-treated. Clinical variables were collected. Mice were euthanized by carbon dioxide inhalation at 8, 12 and 16 weeks of age, and kidneys were removed. Mesangial expansion was analyzed through 10-15 consecutive glomeruli per mouse using ImageJ software. Immunostaining for MMP-10 was performed. Renal tissue RNA was extracted. MMP-10 and b-actin genes were assessed. Differences between groups were analyzed using ANOVA, followed by post-hoc tests, and associations by Pearson correlation. RESULTS: Clinical characteristics of mice are shown in Table 1. At 16 weeks-old, glomerular hypertrophy and mesangial matrix expansion were increased in db/db mice (Figure 1.A-D). Eight, 12 and 16 weeks-old, db/db mice showed increased renal MMP- 10 compared to controls (Figure 2.A). Renal MMP-10 was observed in podocytes and juxtaglomerular apparatus (Figure 2.B-E.). Treatment with Telmisartan significantly downregulated MMP-10 gene expression after 4 (p<0.01) and 8 weeks (p<0.05). A positive correlation was observed between renal MMP-10 gene expression and albuminuria in db/db (R=0.53; p=0.02). i510 | Abstracts Abstracts Nephrology Dialysis Transplantation Downloaded from https://academic.oup.com/ndt/article/34/Supplement_1/gfz103.SP452/5515183 by guest on 22 February 2023