Lipid deposits can injury the kidney of diabetic patients and models. Sterol regulatory element binding protein-1 (SREBP-1) is transcription factor regulating the synthesis of fatty acid and triglyceride. At present whether the expression of SREBP-1 makes some effects on the lipid accumulation in diabetic kidney is not still clear completely. The purpose of our in vivo and in vitro study is to investigate the relationship between the expression of SREBP-1 and lipid abnormal metabolism in the type 1 diabetic rats and explore to inhibit SREBP-1 gene expression by RNA interfere in human renal proximal tubular epithelial cells line (HKC cells). The animal experiment showed that triglyceride and SREBP-1 were up-regulated in proximal tubule of diabetic rats' kidney, which may result in increase of transforming growth factor-beta1 (TGF-beta1) and accumulation of extracellular matrix (ECM). The further HKC cells experiment confirmed SREBP-1 increasing resulted into lipid droplet formation. The expression of fatty acid synthase (FAS) in HKC cells transfected with specific plasmid for SREBP-1 gene was significantly more than that of the cells transfected with the control plasmid pcDNA3.1 and that of the untransfected cells. Simultaneously, up-regulation of TGF-beta1 and fibronectin, an ECM glycoprotein, was evident in HKC cells transfected by specific SREBP-1 plasmid. Furthermore, we found that high glucose was a positive factor on the expression of SREBP-1 at protein and mRNA levels in HKC cells. High glucose makes effects on SREBP-1 in time-dependent manner, and the greatest effect was at 48 h. In addition, two effective eukaryotic expression plasmid vectors of shRNA aimed at SREBP-1 were designed and constructed successfully. Compared with the negative control plasmid group, the levels of the expression of SREBP-1 were inhibited by 24.11 and 36.15%, respectively, at mRNA level, 20.80 and 37.59%, respectively, at precursor segment of protein level, and 38.12 and 52.24%, respectively, at mature segment of protein level at 48 h after transfection. In vivo and in vitro study suggested that high glucose caused increasing SREBP-1 mRNA and protein in renal proximal tubule epithelial cells of type 1 diabetic rats. Increasing SREBP-1 plays an important role in the pathogenesis of renal lipid accumulation by up-regulation of FAS and ECM accumulation by inducing TGF-beta1 expression. The application of vector-mediated RNAi could markedly inhibit the expression of SREBP-1 in HKC cells, which is a promising tool for future research into the mechanisms of renal lipid accumulation in vivo.

BackgroundNephronophthisis-related ciliopathies (NPHP-RC) account for the majority of cases of monogenetically caused end-stage renal disease (ESRD) in children. Exploring the correlation between the phenotype and genotype of NPHP-RC is helpful for early diagnosis and management. We investigated the phenotype and genotype spectra of NPHP-RC in a Chinese multicentre cohort.MethodsCrosss-ectional and longitudinal data of 60 patients from 57 families with pathogenic NPHP-RC gene mutations distributed in 22 regions of China were collected into a unified, anonymous database. The mean observation time of this cohort was 3.5±3.1 years.ResultsMutations in NPHP1 and NPHP3 were the most common genetic defects. Overall, 45% of patients presented with isolated nephronophthisis (NPH), and 55% exhibited the extrarenal phenotype, which frequently involved the liver (41.7%, n=25), central nervous system (26.7%, n=16), eyes (26.7%, n=16) and skeletal system (11.7%, n=7). Accidental detection of elevated serum creatinine and non-specific symptoms caused by chronic kidney disease occurred in 65% of patients. Patients carrying NPHP1 mutations mainly presented with isolated NPH (90%, 18/20) and progressed to ESRD at a mean age of 12.9±0.5 years. The mean age of ESRD onset in the non-NPHP1 group was lower than that in the NPHP1 group (6.2±1.4 years, p<0.001), especially for patients carrying NPHP3 mutations (3.1±1.2 years), showing a heterogeneous phenotype characterised by Bardet-Biedl syndrome (12.5%, n=5), Joubert syndrome (7.5%, n=3), COACH syndrome (2.5%, n=1), Mainzer-Saldino syndrome (2.5%, n=1), short-rib thoracic dysplasia (2.5%, n=1) and unclassified symptoms (32.5%, n=13).ConclusionsThe Chinese Children Genetic Kidney Disease Database registry characterised the spectrum of the phenotype and genotype of NPHP-RC in the Chinese population. NPHP1 and NPHP3 were the most common pathogenic genes. Rapid progression to ESRD and liver involvement were noted in patients with NPHP3 mutations.

In the present study, the effect and mechanism of periostin on renal proliferation and extracellular matrix accumulation of lupus mice were investigated. MRL /lpr mice, known as lupus mice, were revealed to show enhanced periostin, proliferating cell nuclear antigen (PCNA), and extracellular matrix accumulation in the kidney accompanied by increased serum platelet-derived growth factor (PDGF). Again, cultured mouse mesangial cells (MMCs) were treated with PDGF, then periostin, and PCNA and secreted fibronectin were detected. The results showed that intracellular periostin and PCNA were respectively enhanced by 2.691 and 2.308 times in PDGF-treated MMC cells at 6 h after stimulation. In addition, secreted fibronectin was increased by 1.442 times. Next, the transfection of periostin shRNA vector in PDGF-stimulated MMC cells effectively suppressed periostin, PCNA and secreted fibronectin by 45.27%, 47.75%, and 39.95%, compared with PDGF-stimulated cells transfected with control vector. Furthermore, it was found that PDGF increased the expression of phospho-Akt (Ser 473) from 30 min to 6 h in MMCs. LY294002 effectively inhibited phospho-Akt (Ser 473) expression caused by PDGF stimulation. Then, periostin, PCNA, and fibronectin were respectively decreased by 69.61%, 46.00%, and 46.20%. In the end, phosphoinositide 3-kinase/protein kinase B/periostin was suggested to mediate PDGF-induced cell proliferation and extracellular matrix production in lupus nephritis.

Background: To observe the changes of autophagy-related protein levels in peripheral blood lymphocytes before and after sirolimus treatment in children with systemic lupus erythematosus (SLE).Methods: Children with SLE were randomly divided into two groups, 28 in the traditional treatment group and 28 in the sirolimus group. Fifteen healthy children who were in the same period were collected as the normal control group. Clinical laboratory indexes, the percentage of routine lymphocytes, complement C3, complement C4, serum Anti-dsDNA and SLEDAI were detected.Results: At 3 and 6 months after treatment, compared with the traditional treatment group, the percentage of routine lymphocytes in the sirolimus group increased (P = 0.03), SLEDAI score and positive rate of Anti-dsDNA decreased (P = 0.01). Compared with normal children, the expression of microtubule-associated protein 1 light chain 3 (LC3) protein in peripheral blood lymphocytes was significantly higher (P = 0.006); peripheral blood expression of P62/SQSTM1 (sequestosome 1) protein in lymphocytes decreased (P = 0.02).Conclusion: Sirolimus can play a role in the treatment of systemic lupus erythematosus by regulating the level of autophagy.

Objective: To identify the gene mutation of Stormorken syndrome and review the published Stromal Interaction Molecule 1 (STIM1) mutation phenotype.Methods: We described the clinical and molecular aspects of a Chinese female with Stormorken syndrome by laboratory tests, muscle biopsies, and genetic analysis. We used this information to summarize all the mutation sites reported in the literature. We also reviewed the clinical features of published cases with a gain of function mutations of STIM1.Results: A 12-year-old Chinese female presented with skin purpura in the lower limbs and stroke-like episodes. Muscle biopsy and microscopic examination revealed atrophy in her skeletal muscle. Genetic analysis identified a novel heterozygous missense mutation, a c.1095G&gt;C transition (NM_003156.3), which caused a p.K365N amino acid substitution in the protein and affected a STIM1-orai1-activation region (SOAR).Conclusions: The novel variant c.1095G&gt;C transition (NM_003156.3) was located in the SOAR, which expands the phenotypic spectrum of STIM1 variants in human disorders and may define the molecular basis of Stormorken syndrome.