cytometry techniques. Mitochondrial b-oxidation rate was examined by commercial kits. RESULTS: The results showed that high glucose (30 mM) was shown to induce increased expression of NLRP3, IL-1b and caspase-1. This was blocked by knockdown of CD36 or antioxidant TEMPO. Meanwhile, we also found that knockdown of CD36 inhibited high glucose-induced generation of mitochondrial ROS, a–SMA, TGF-b1 and increased levels of E-cadherin, p-AMPK, mitochondrial b-oxidation. CONCLUSIONS: These results suggested that knockdown of CD36 antagonized high glucose-induced NLRP3 inflammasome activation by upregulating mitochondrial fatty acid oxidation and inhibiting ROS production, CD36 as a potential therapy target for diabetic nephropathy. SP445 A1AR ALLEVIATED MICROVASCULAR LOSS VIA INHIBITING PERICYTES DETACHMENT INDUCED EXTRACELLULAR MATRIX ACCUMULATION IN DIABETIC NEPHROPATHY Dongli Tian 1 , Linfeng Zou 1 , Min Lin 1 , Xiaoxiao Shi 1 , Lubin Xu 1 , Peng Xia 1 , Yubing Wen 1 , Limeng Chen 1 1 Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing, China INTRODUCTION: We previously observed A1 adenosine receptor (A1AR) had a protective role in the progression of diabetic nephropathy. But the mechanism was unclear. In this study, we used A1AR deficient mice with diabetic nephropathy to prove whether A1AR play a role in the pericyte detachment mediated peritubular capillary injury and in the progress of renal interstitialfibrosis in type 1 diabetic nephropathy. METHODS: Successfully established streptozotocin induced type 1 diabetic nephropathy model in wild and A1AR deficient mice for 4 and 16 weeks. Urine, blood and kidney tissues were collected. Renal pathological lesion was evaluated by light and electron microscopes. Using mRNA expression profile to screening differentially expressed genes, and verifying the results by western blot to detect the protein expression. RESULTS: Compared to 4 weeks, macroabuminuria and serious renal pathological leisure evaluated by HE and Masson staining were observed in wild diabetic mice at 16 weeks, with obvious up-regulation of ten mRNA expression including pericyte detachment induced platelet-derived growth factor receptor (PDGFRb, 1.5times, P=0.023) activation, peritubular capillary loss (anti-CD34 antigen, 0.5times, P=0.001), A1AR (1.3times, P=0.042) and collagen related family mRNA detected by mRNA expression profile. While in A1AR deficient diabetic mice, more severe renal tubular interstitial fibrosis and more extracellular matrix deposition evaluated by electron microscope, with more increased protein expression of PDGFRb (1.6times, P=0.008) and lower CD34 (0.2times, P<0.001) protein expression than that in wild diabetic mice detected by immunoblotting. Also with higher protein expression of transforming growth factor (TGFb, 3.0times, P<0.001), collagen I (2.2times, P<0.001), collagen III (2.0times, P=0.006), collagen IV (1.8times, P<0.001) than in wild diabetic mice. CONCLUSIONS: A1AR plays a protective role in the PDGFR-mediated peritubular capillary injury to alleviate renal extracellular matrix deposition and tubular interstitial fibrosis in type 1 diabetic nephropathy. SP446 ACE2 DELETION IN NOD MICE INCREASES FIBROSIS IN PRIMARY PROXIMAL TUBULAR CELLS Vanesa Palau 1 , Marta Riera 1 , David Benito 1 , Julio Pascual 1 , Maria Jose ´ Soler 2 1 Institut Hospital del Mar d’Investigacions Me `diques IMIM, Barcelona, Spain and 2 Institut Hospital del Mar d’Investigacions Me `diques IMIM. pi17/00257, Barcelona, Spain INTRODUCTION: Angiotensin converting enzyme (ACE) 2 acts as a negative regulator of the renin angiotensin system. It has been demonstrated that ACE2 pharmacology inhibition or genetic deletion worsens renal lesions. Our aim was to study the effect of ACE2 deletion on fibrosis as a marker of renal damage in primary proximal tubular cells from NOD mice in a pre-diabetic state. METHODS: NOD-ACE2-/- and NOD-ACE2+/+ female mice at 12 weeks of age were analyzed. Renal cortex was digested in culture media with collagenase type II. Cells separation was performed through to 100mm- and 80mm-sieves. Obtained cells were cultured in DMEM-F12 media and sub-cultured for 4 times. Cells characterization was performed by SGLT-2 and AQP1 gene expression as positive markers and AQP2 gene expression as a negative marker. Gene expression of collagen-I, collagen-IV, fibronectin, and a-smooth muscle actin (SMA) was determined by qPCR. RESULTS: As expected, RNA isolated from primary proximal tubular cells wild-type and knockout for ACE2 were positive for SGLT-2 and AQP1 gene expression and negative for AQP2 gene expression. Interestingly, ACE2 knockout cells presented increased collagen-IV and a-SMA gene expression as compared to wild-type cells. CONCLUSIONS: ACE2 deletion in proximal tubular cells may contribute to renal fibrosis. The lack of ACE2 may result in the accumulation of angiotensin II (ANGII) and the stimulation of the ANGII/AT1R axis and subsequent tubular damage. SP447 INTEGRIN-LINKED KINASE (ILK) IS DECREASED DURING THE DEVELOPMENT OF OBESITY IN A SHORT-TERM HIGH FAT DIET MICE MODEL Griera Mercedes 1 , Hatem-Vaquero Marco 1 , Garc  ıa-Ayuso Diego 1 , Campillo Sofia 2 , Bohorquez Lourdes 2 , Olmos Gemma 2 , M. Piedad Ruiz-Torres 1 , Lopez-Ongil Susana 3 , Rodr  ıguez-Puyol Diego 3 , Rodr  ıguez-Puyol Manuel 2 , Sergio de Frutos 1 , Calleros Laura 4 1 Universidad de Alcal a, ALCALA DE HENARES, Spain, 2 Instituto de Salud Carlos III, Madrid, Spain, 3 Hospital Pr  ıncipe de Asturias, Madrid, Spain and 4 Alcala University, Alcal a de Henares, Madrid, Spain INTRODUCTION: Development of obesity and dysregulation of lipolysis in visceral white adipose tissue (WAT) are identified important mechanisms in the pathophysiology associated with the establishment of insulin resistance Obesity can be understood as the increase in WAT deposit weights. The dysregulation of lipolysis in visceral WAT can be observed by changes in the expression and activity of the hormone-sensitive lipase (HSL), the major lipase involved in hormone-regulated lipolysis inside WAT, and the determination in the blood of the lipolysis-end products glycerol and free fatty acids (FFA). [Czech MP, Nat Med, 2017. 23(7): p. 804-814]. However, the precise mechanisms underpinning the metabolites disposal on key metabolic organs remains unrevealed. We recently published that depletion of the scaffold intracellular protein Integrin-linked kinase (ILK) is a modulator during insulin resistance in key organs such as WAT [Hatem-Vaquero et al. J Endocrinol. 2017; 234(2):115-128]. Here we used a transgenic mice model of global ILK depletion (cKD-ILK) and their wildtype (WT) counterparts subjected to a high fat diet (HFD) challenge for a short period of time, which allows to investigate the time course of the obesity and dyslipidemia implementation [Park et al. Diabetes 54: 3530–3540, 2005]. Under these conditions we compared the body weight changes, and the lipolysis dysregulations and ILK expression in their visceral (epididymal) WAT (eWAT). METHODS: Transgenically modified mice [Hatem-Vaquero et al. J Endocrinol. 2017; 234(2):115-128] carrying the floxed ILK allele and CMV-driven tamoxifen-inducible CreER (T) recombinase gene were used for the experiments in their adulthood. Those that displayed a successful ILK loss (cKD-ILK) in the insulin-sensitive organs, one month after tamoxifen induction of ILK depletion, mice and their corresponding wild- type littermates (WT), were fed either with a standard chow diet (time 0) or HFD (60% calories from fat lard, D12492 Ssniff Spezialdi€ aten) for 2 and 6 weeks. Differences between groups (WT or cKD-ILK fed with chow diet or HFD for 2 and 6 weeks of total body and eWAT deposits weights, eWAT mRNA and protein ILK, HSL and blood glycerol and FFA levels were determined. RESULTS: ILK expression in eWAT under HFD challenge significantly decreased as early as 2 weeks, reaching the same constantly low levels of cKD-ILK from time 0 to the 6th week. The eWAT lipolysis pattern of cKD-ILK and WT at time 0 was unaltered. However HSL expression after 2 weeks of HFD significantly increased in WT and cKD- ILK, and the last reached the highest values. However the 6th week HSL expression decreased to basal levels in both groups. On parallel, HFD exposure resulted in increased blood glycerol levels as early as 2 weeks, where the cKD-ILK reached the highest levels. At the 6th week, both WT and cKD-ILK maintained high levels. No changes in FFA blood levels were observed between groups. CONCLUSIONS: We suggest ILK as a modulator during the obesity establishment. ILK expression may be a potential predictive value and taking in consideration the relation between ILK and the lipolysis pattern observed here, the upregulation of ILK expression may be a potential target to prevent dyslipidemia. SP448 ROLE OF ACUTE-PHASE PROTEINS (ORSOMUCOID 2 AND HS-CRP) IN LEAN TYPE 2 DIABETIC PATIENTS WITH AND WITHOUT SUBCLINICAL ATHEROSCLEROSIS Maha Salama 1 1 Alzhraa hosiptal- Alazhr universty, cario, Egypt INTRODUCTION: Background: Type 2 Diabetes Mellitus (T2DM), is a global public health crisis that threatens the economies of all nations, particularly developing countries. Cardiovascular complications are the main cause of mortality in T2DM patients. Objective: The aim of this study is to explore the role of acute-phase proteins (Orsomucoid 2 and hs-CRP) as predictors for subclinical atherosclerosis and their relation with dyslipidaemia and Carotid intima media thicknes (CIMT) in lean T2DM patients. METHODS: 50 diabetic normal weight patients (22 males and 28 females) were recruited from internal medicine department, AL-Zahraa hospital and divided into two groups: Group 1; twenty five type 2 diabetics with increased CIMT ( 0.9 mm) and Group 2; twenty five diabetic patients with normal CIMT (<0.9 mm). All the participants were subjected to full medical history, full clinical examination, and routine Laboratory investigations: Serum fasting glucose, urea, creatinine, uric acid, doi:10.1093/ndt/gfz103 | i509 Nephrology Dialysis Transplantation Abstracts Downloaded from https://academic.oup.com/ndt/article/34/Supplement_1/gfz103.SP447/5515139 by guest on 29 April 2022