目的： 转化生长因子－1（TGF-β1）通过与II型受体结合而引起I型受体活化，进一步激活其下游信号分子蛋白Smad2 和Smad3，它们与Smad4（Co-Smad）结合后形成Smad复合体并发生核转移，从而发挥广泛的生物学效应。同时，整个TGF-β信号通路又受到其抑制因子Smad7的负反馈调节。研究结果显示Smad3是肾脏炎症和纤维化中重要的致病分子，相反，Smad7在多种肾脏疾病中起保护作用。然而，由于转化生长因子II型受体（TβRII），Smad2 或Smad4基因敲除的小鼠无法存活，这些分子在TGF-β1介导的肾脏炎症和纤维化中的功能尚未见报道。因此，本研究旨在剖析TβRII、Smad2 和Smad4 在肾脏疾病发生发展中的作用及机制。 / 方法：本研究利用Cre/LoxP系统分别靶向敲除小鼠肾小管上皮细胞的TβRII、Smad2 或者Smad4，通过结扎小鼠单侧输尿管建立梗阻性肾病模型，观察这些分子对肾脏炎症和纤维化的影响，并用体外实验进行验证。具体实验结果请参见本论文第III,IV, V章。 / 结果：通过分析，本论文取得以下新的发现： / (1) TβRII在TGF-β1介导的肾脏炎症和纤维化的双向调节中起到了决定性的作用：研究结果显示条件性敲除TβRII明显抑制TGF-β/Smad3介导的肾脏纤维化，同时增强NF-κB引起的肾脏炎症反应。由此可见，TRII不仅仅是TGF-β/Smad信号通路的启动因子，更决定了TGF-β1对肾脏炎症和纤维化的双向性调节。(参见第III章) / (2)尽管Smad2和Smad3结构相似并共同介导了TGF-β1的生物学效应，本研究意外发现Smad2可反向调节Smad3引起的纤维化。体内和体外实验共同证实，敲除Smad2基因增强了Smad3的磷酸化，核转位及其转录子活性，并能促进Smad3与I型胶原转录子的结合，进而加重肾脏纤维化（参见第IV章）。 / (3)我们还发现Smad4不仅作为TGF-β/Smad信号通路的共有蛋白，它在TGF-β1介导肾脏炎症和纤维化中起到了重要的双向性调节作用：条件敲除Smad4显著降低了Smad7对NF-κB介导肾脏炎症的抑制作用，同时在转录水平（而非磷酸化水平）抑制Smad3的功能，从而减轻纤维化。（参见第V章） / 结论：TβRII和Smad4 在TGF-β1介导肾脏炎症和纤维化中起到了重要的双向性作用；Smad2通过抑制Smad3信号传导和功能，在肾脏纤维化中起保护作用。 / Objectives: TGF-β1 binds its receptor II (TβRII) and then activates receptor I to initiate the downstream Smad signaling, called Smad2 and Smad3 which bind a common Smad4 to form the Smad complex and then translocate to nucleus to exert its biological activities. This process is negatively regulated by an inhibitory Smad7. While the pathogenic role of Smad3 and the protective role of Smad7 in renal fibrosis and inflammation are clearly understood, the functional role of TβRII, Smad2 and Smad4 in kidney diseases remains largely unexplored due to the lethality of these knockout mice. Therefore, the aim of present study is to dissect the functional role of these TGF-β/Smad signaling molecules in renal inflammation and fibrosis. / Methods: Kidney conditional knockout (KO) mice for TβRII, Smad2 and Smad4 were generated by crossing the FloxFlox mice with the kidney specific promoter driven Cre (KspCre) mice, in which TβRII, Smad2 or Smad4 were specifically deleted from the kidney tubular epithelial cells (TEC) respectively. Then, a well-characterized progressive renal inflammation and fibrosis mouse model of Unilateral ureteral obstructive (UUO) nephropathy was induced in these conditional KO mice and the specific roles for TβRII, Smad2, and Smad4 in renal inflammation and fibrosis were investigated in vivo and in vitro as described in the Chapter III, IV and V of this thesis. / Results: There were several novel findings through this thesis: / 1. TGF-β1 signals through its TβRII to diversely regulate renal fibrosis and inflammation. We found that disrupted TRII suppressed Smad3-dependent renal fibrosis while enhancing NF-κB-driven renal inflammation. Thus, TβRII not only acts as a binding receptor for initiating the TGF-β signaling, but also determines the diverse role of TGF-β1 in inflammation and fibrosis, which was described in the Chapter III. / 2. As shown in the Chapter IV, an unexpected finding from this thesis was that although Smad2 and Smad3 were homologically similar and bound together in response to TGF-β1 stimulation, Smad2 counter-regulated Smad3-mediated renal fibrosis. This was evidenced by the findings that conditional deletion of Smad2 enhanced Smad3 signaling including phosphorylation, nuclear translocation, the Smad3 responsive promoter activity, and the binding of Smad3 to Col1A2 promoter. Thus, disrupted Smad2 from the kidney significantly enhanced Smad3-mediated renal fibrosis in the UUO kidney and in cultured TEC. / 3. Finally, we also showed that that Smad4 acted not only as a common Smad in TGF-β signaling, but exerted its regulatory role in determining the diverse role of TGF-β1 in renal inflammation and fibrosis. Disruption of Smad4 significantly enhanced renal inflammation by impairing inhibitory effect of Smad7 on NF-κB-driven renal inflammation. In contrast, disrupted Smad4 inhibited renal fibrosis by blocking Smad3 functional activity without influencing Smad3 signaling. Because deletion of Smad4 inhibited TGF-β1-induced Smad3 responsive promoter activity and the binding of Smad3 to the Col1A2 promoter without altering the phosphorylation and nuclear translocation of Smad3 (Chapter V). / Conclusions: TβRII and Smad4 may function as key regulators of TGF-β signaling and diversely regulate the renal inflammation and fibrosis. Smad2 plays a protective role in renal fibrosis by counter-regulating Smad3 signaling. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Meng, Xiaoming. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 202-231). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / Abstract --- p.i / Declaration --- p.viii / Acknowledgement --- p.ix / Table of Contents --- p.xii / List of Abbreviations --- p.xxvii / List of Figures/Tables --- p.xxix / Chapter CHAPTER I --- INTRODUCTION --- p.1 / Chapter 1.1 --- TGF-β signaling pathway --- p.2 / Chapter 1.1.1 --- TGF-β superfamily --- p.2 / Chapter 1.1.2 --- TGF-β signaling transduction --- p.3 / Chapter 1.1.2.1 --- Smad-dependent TGF-β signaling --- p.4 / Chapter 1.1.2.2 --- Smad-independent TGF-β signaling --- p.10 / Chapter 1.2 --- Chronic Kideny disease (CKD) --- p.12 / Chapter 1.2.1 --- Epidemiology of CKD --- p.12 / Chapter 1.2.2 --- Pathophysiology of CKD --- p.12 / Chapter 1.3 --- TGF-β signaling in renal diseases --- p.13 / Chapter 1.3.1 --- Role of TGF-β1 in renal diseases --- p.13 / Chapter 1.3.2 --- Potential role of TβRII in renal diseases --- p.15 / Chapter 1.3.3 --- Potential role of Smad2 in renal diseases --- p.17 / Chapter 1.3.4 --- Potential role of Smad4 in renal diseases --- p.20 / Chapter 1.3.5 --- Role of Smad7 in renal diseases --- p.23 / Chapter 1.3.6 --- Role of Smad-independent TGF-β signaling in renal disease --- p.24 / Chapter CHAPTER II --- MATERIALS AND METHODS --- p.26 / Chapter 2.1 --- MATERIALS --- p.27 / Chapter 2.1.1 --- Reagents and Equipments --- p.27 / Chapter 2.1.1.1 --- General reagents and equipments for cell culture --- p.27 / Chapter 2.1.1.2 --- General reagents and equipments for real-time RT-PCR --- p.28 / Chapter 2.1.1.3 --- General reagents and equipments for Masson Trichrome Staining --- p.28 / Chapter 2.1.1.4 --- General reagents and equipments for Immunohistochemistry --- p.29 / Chapter 2.1.1.5 --- General reagents and equipments for Immunofluorescence --- p.29 / Chapter 2.1.1.6 --- General reagents and equipments for Western Blot --- p.29 / Chapter 2.1.1.7 --- General reagents and equipments for Promoter assay --- p.31 / Chapter 2.1.1.8 --- General reagents and equipments for ChIP assay --- p.32 / Chapter 2.1.2 --- Buffers --- p.32 / Chapter 2.1.2.1 --- Buffers for Immunohistochemistry --- p.32 / Chapter 2.1.2.2 --- Buffers for Western blot --- p.35 / Chapter 2.1.3 --- Sequences of Primers and siRNAs --- p.40 / Chapter 2.1.4 --- Antibodies --- p.42 / Chapter 2.2 --- METHODS --- p.44 / Chapter 2.2.1 --- Animal model of Unilateral Ureteral Obstruction (UUO) --- p.44 / Chapter 2.2.2 --- Cell culture --- p.44 / Chapter 2.2.2.1 --- NRK52E cell line --- p.44 / Chapter 2.2.2.2 --- Smad2 WT/KO mouse embryonic fibroblasts (MEFs) --- p.45 / Chapter 2.2.2.3 --- Primary culture of kidney fibroblasts --- p.45 / Chapter 2.2.2.4 --- Primary culture of peritoneal macrophages --- p.46 / Chapter 2.2.3 --- PAS staining --- p.47 / Chapter 2.2.3.1 --- Tissue Handling and Fixation --- p.47 / Chapter 2.2.3.2 --- Tissue embedding and sectioning --- p.47 / Chapter 2.2.3.3 --- Preparation of Paraffin Tissue Sections for PAS staining --- p.48 / Chapter 2.2.3.4 --- PAS staining --- p.48 / Chapter 2.2.4 --- Real-time RT-PCR --- p.48 / Chapter 2.2.4.1 --- Total RNA isolation --- p.48 / Chapter 2.2.4.2 --- Reverse Transcription --- p.49 / Chapter 2.2.4.3 --- Real-time PCR --- p.50 / Chapter 2.2.4.4 --- Analysis of Real-time PCR --- p.50 / Chapter 2.2.5 --- Masson Trichrome Staining --- p.51 / Chapter 2.2.6 --- Immunohistochemistry --- p.52 / Chapter 2.2.6.1 --- Preparation of Paraffin Tissue Sections for IHC --- p.52 / Chapter 2.2.6.2 --- Antigen-Antibody Reaction --- p.52 / Chapter 2.2.6.3 --- Signal Detection --- p.53 / Chapter 2.2.6.4 --- Semi-quantification of Immunohistochemistry --- p.53 / Chapter 2.2.7 --- Immunofluorescence --- p.54 / Chapter 2.2.8 --- Western blot analysis --- p.54 / Chapter 2.2.8.1 --- Protein preparation --- p.55 / Chapter 2.2.8.2 --- SDS-PAGE --- p.56 / Chapter 2.2.8.3 --- Transmembrane of protein --- p.56 / Chapter 2.2.8.4 --- Incubation of first and second antibody --- p.57 / Chapter 2.2.8.5 --- Signal capture and analysis --- p.57 / Chapter 2.2.8.6 --- Stripping --- p.57 / Chapter 2.2.9 --- Promoter assay --- p.58 / Chapter 2.2.10 --- ChIP assay --- p.61 / Chapter 2.2.11 --- Statistical analysis --- p.62 / Chapter CHAPTER III --- THE DIVERSE ROLE OF TGF-BETA RECEPTOR II IN RENAL INFLAMMATION AND FIBROSIS --- p.63 / Chapter 3.1 --- INTRODUCTION --- p.64 / Chapter 3.2 --- AIMS --- p.64 / Chapter 3.3 --- MATERIALS AND METHODS --- p.66 / Chapter 3.3.1 --- Generation and characterization of TβRII conditional Knockout mice --- p.66 / Chapter 3.3.2 --- Generation and characterization of TβRII disrupted tubular epithelial cell line (NRK52E) and kidney interstitial fibroblasts --- p.67 / Chapter 3.3.3 --- Animal model of Unilateral Ureteral Obstruction --- p.67 / Chapter 3.3.4 --- Cell culture --- p.67 / Chapter 3.3.5 --- Real-time RT-PCR --- p.68 / Chapter 3.3.6 --- Masson Trichrome Staining --- p.68 / Chapter 3.3.7 --- Immunohistochemistry --- p.68 / Chapter 3.3.8 --- PAS staining --- p.69 / Chapter 3.3.9 --- Immunofluorescence --- p.69 / Chapter 3.3.10 --- Western blot analysis --- p.70 / Chapter 3.3.11 --- Promoter assay --- p.70 / Chapter 3.3.12 --- Statistical analysis --- p.70 / Chapter 3.4 --- RESULTS --- p.71 / Chapter 3.4.1 --- Characterization of TβRII conditional Knockout mice and TβRII disrupted cells --- p.71 / Chapter 3.4.2 --- Disruption of TβRII suppresses renal interstitial damage in the UUO kidney --- p.72 / Chapter 3.4.3 --- Disruption of TβRII suppresses renal fibrosis in UUO kidney and TGF-β1-induced fibrotic response in vitro --- p.76 / Chapter 3.4.3.1 --- Conditional knockout of TβRII from the kidney decreases the collagen I level in UUO kidney --- p.76 / Chapter 3.4.3.2 --- Disruption of TβRII inhibits TGF-β1 induced collagen I level in vitro --- p.79 / Chapter 3.4.3.3 --- Conditional knockout of TβRII from the kidney decreases the α-SMA positive cells infiltration in vivo --- p.81 / Chapter 3.4.3.4 --- Disruption of TβRII inhibits TGF-β1-induced α-SMA expression in vitro --- p.83 / Chapter 3.4.3.5 --- Conditional knockout of TβRII from the kidney decreases the FN level in UUO nephropathy --- p.85 / Chapter 3.4.3.6 --- Disruption of TβRII decreases TGF-β1-induced FN expression in vitro --- p.87 / Chapter 3.4.4 --- Disruption of TβRII impairs the TGF-β/Smad signaling in vivo in the UUO kidney and in vitro in TGF-β1 treated tubular epithelial cells and kidney fibroblasts --- p.89 / Chapter 3.4.4.1 --- Conditional knockout of TβRII decreases the UUO induced TGF-β1 expression in vivo and the TGF-β1 auto-induction in vitro --- p.89 / Chapter 3.4.4.2 --- Disrupted TβRII decreases CTGF level in the UUO nephropathy in vivo and the TGF-β1 induced CTGF mRNA level in vitro --- p.91 / Chapter 3.4.4.3 --- Conditional knockout of TβRII impairs the Smad3 signaling in the injured kidney --- p.93 / Chapter 3.4.4.4 --- Disrupted TβRII inhibits TGF-β1-induced Smad3 phosphorylation, P-Smad3 nuclear translocation and Smad3 responsive promoter activity in vitro --- p.95 / Chapter 3.4.4.5 --- Conditional knockout of TβRII doesn’t alter the activation of ERK and P38 signaling in the UUO kidney --- p.97 / Chapter 3.4.4.6 --- Disrupted TβRII inhibits TGF-β1-induced ERK and P38 phosphorylation in vitro --- p.99 / Chapter 3.4.5 --- Disruption of TβRII enhances inflammatory cytokines expression in the UUO kidney and impairs the anti-inflammatory effect of TGF-β1 in response to IL-1β triggered inflammatory response in the TEC cells --- p.101 / Chapter 3.4.5.1 --- Conditional knockout of TβRII increases the TNF-α expression in the UUO nephropathy --- p.101 / Chapter 3.4.5.2 --- Conditional knockout of TβRII increases the IL-1β expression in the UUO nephropathy --- p.103 / Chapter 3.4.5.3 --- Conditional knockout of TβRII doesn’t enhance the MCP-1 expression and macrophages infiltration in the UUO nephropathy --- p.104 / Chapter 3.4.5.4 --- Disruption of TβRII in TECs decreases the anti-inflammatory effect of TGF-β1 in response to IL-1β --- p.106 / Chapter 3.4.6 --- Disruption of TβRII enhances NFκB activation in vivo and in vitro --- p.108 / Chapter 3.5 --- DISCUSSION --- p.110 / Chapter 3.6 --- CONCLUSION --- p.114 / Chapter CHAPTER IV --- Smad2 protects against TGF-β/Smad3 mediated renal fibrosis --- p.115 / Chapter 4.1 --- INTRODUCTION --- p.116 / Chapter 4.2 --- AIMS --- p.117 / Chapter 4.3 --- MATERIALS AND METHODS --- p.117 / Chapter 4.3.1 --- Generation and characterization of Smad2 conditional Knockout mice --- p.117 / Chapter 4.3.2 --- Generation and characterization of Smad2 KO MEFs and Smad2 knockdown/overexpression tubular epithelial cell line (NRK52E) --- p.118 / Chapter 4.3.3 --- Animal model of Unilateral Ureteral Obstruction --- p.118 / Chapter 4.3.4 --- Cell culture --- p.118 / Chapter 4.3.5 --- Real-time RT-PCR --- p.119 / Chapter 4.3.6 --- Western blot analysis --- p.119 / Chapter 4.3.7 --- Immunohistochemistry --- p.119 / Chapter 4.3.8 --- Masson Trichrome Staining --- p.119 / Chapter 4.3.9 --- Immunofluorescence --- p.120 / Chapter 4.3.10 --- Promoter assay --- p.120 / Chapter 4.3.11 --- ChIP assay --- p.120 / Chapter 4.3.12 --- Statistical analysis --- p.120 / Chapter 4.4 --- RESULTS --- p.121 / Chapter 4.4.1 --- Characterization of Smad2 disrupted mice and cells --- p.121 / Chapter 4.4.1.1 --- Characterization of Smad2 conditional Knockout mice --- p.121 / Chapter 4.4.1.2 --- Characterization of Smad2 knockout MEFs, Smad2 knockdown/overexpression TECs --- p.123 / Chapter 4.4.2 --- Disruption of Smad2 further enhances renal fibrosis in vivo and in vitro --- p.124 / Chapter 4.4.2.1 --- Conditional knockout of Smad2 increases total collagen deposition and Col.I level in the UUO kidney --- p.124 / Chapter 4.4.2.2 --- Disruption of Smad2 in MEFs and TECs increases Col.I production in a time- and dosage-dependent manner in response to TGF-β1 --- p.126 / Chapter 4.4.2.3 --- Conditional knockout of Smad2 increases Col.III level in the UUO kidney --- p.128 / Chapter 4.4.2.4 --- Disruption of Smad2 in MEFs and TECs increases Col.III production in a time- and dosage-dependent manner in response to TGF-β1 --- p.130 / Chapter 4.4.3 --- Disruption of Smad2 further enhances renal fibrosis by suppressing the collagen degradation system in vivo and in vitro --- p.132 / Chapter 4.4.3.1 --- Conditional knockout of Smad2 inhibits the MMP2 mRNA while enhances TIMP-1 production in UUO kidney --- p.132 / Chapter 4.4.3.2 --- Disruption of Smad2 in MEFs and TECs decreases the MMP2 level while enhances TIMP-1 production in response to TGF-β1 --- p.133 / Chapter 4.4.4 --- Disruption of Smad2 further increases renal fibrosis by increasing TGF-β1 auto-induction and CTGF level in vivo and in vitro --- p.135 / Chapter 4.4.4.1 --- Disruption of Smad2 increases TGF-β1 auto-induction in vivo and in vitro --- p.135 / Chapter 4.4.4.2 --- Disruption of Smad2 increases CTGF synthesis in vivo and in vitro --- p.137 / Chapter 4.4.5 --- Disruption of Smad2 further increases renal fibrosis by enhancing Smad3 signaling in vivo and in vitro --- p.139 / Chapter 4.4.5.1 --- Conditional knockout of Smad2 further enhances Smad3 phosphorylation and nuclear translocation --- p.139 / Chapter 4.4.5.2 --- Disruption of Smad2 in MEFs and TECs further enhances Smad3 phosphorylation, nuclear translocation, Smad3 responsive promoter activity and the binding to the Col1A2 promoter --- p.141 / Chapter 4.4.6 --- Overexpression of Smad2 suppresses Smad3 signaling therefore ameliorates the TGF-β1-induced fibrotic response in TECs --- p.144 / Chapter 4.4.6.1 --- Overexpression of Smad2 ameliorates the TGF-β1- induced fibrotic response in TECs --- p.144 / Chapter 4.4.6.2 --- Overexpression of Smad2 suppresses Smad3 phosphorylation --- p.146 / Chapter 4.5 --- DISCUSSION --- p.147 / Chapter 4.6 --- CONCLUSION --- p.150 / Chapter CHAPTER V --- THE DISTINCT ROLE OF SMAD4 IN RENAL INFLAMMATION AND FIBROSIS --- p.151 / Chapter 5.1 --- INTRODUCTION --- p.152 / Chapter 5.2 --- AIMS --- p.152 / Chapter 5.3 --- MATERIALS AND METHODS --- p.153 / Chapter 5.3.1 --- Generation and characterization of Smad4 conditional Knockout mice --- p.153 / Chapter 5.3.2 --- Generation and characterization of Smad4 disrupted kidney interstitial fibroblasts and peritoneal macrophages --- p.153 / Chapter 5.3.3 --- Animal model of Unilateral Ureteral Obstruction (UUO) --- p.154 / Chapter 5.3.4 --- Cell culture --- p.154 / Chapter 5.3.5 --- Real-time RT-PCR --- p.155 / Chapter 5.3.6 --- Western blot analysis --- p.155 / Chapter 5.3.7 --- Immunohistochemistry --- p.155 / Chapter 5.3.8 --- Masson Trichrome Staining --- p.155 / Chapter 5.3.9 --- Promoter assay --- p.156 / Chapter 5.3.10 --- ChIP assay --- p.156 / Chapter 5.3.11 --- Statistical analysis --- p.156 / Chapter 5.4 --- RESULTS --- p.157 / Chapter 5.4.1 --- Characterization of Smad4 conditional Knockout mice and Smad4 disrupted cells --- p.157 / Chapter 5.4.2 --- Disruption of Smad4 suppresses renal fibrosis in the UUO nephropathy in vivo and TGF-β1-induced fibrotic response in vitro --- p.160 / Chapter 5.4.2.1 --- Conditional knockout of Smad4 from the kidney decreases the total collagen deposition in the UUO nephropathy --- p.160 / Chapter 5.4.2.2 --- Conditional knockout of Smad4 from the kidney decreases the Col.I production in the UUO nephropathy --- p.161 / Chapter 5.4.2.3 --- Disruption of Smad4 inhibits TGF-β1-induced Col.I production in vitro --- p.163 / Chapter 5.4.3 --- Disruption of Smad4 impairs the Smad3 function in vivo and in vitro --- p.164 / Chapter 5.4.3.1 --- Conditional knockout of Smad4 doesn’t decrease Smad3 phosphorylation and P-Smad3 nuclear translocation in vivo and in vitro --- p.164 / Chapter 5.4.3.2 --- Disruption of Smad4 inhibits TGF-β1 induced Smad3 promoter activity and the Smad3 binding to Col1A2 promoter --- p.166 / Chapter 5.4.3.3 --- Disruption of Smad4 has minimal effect on the activation of ERK signaling in vivo and in vitro --- p.167 / Chapter 5.4.4 --- Disruption of Smad4 enhances renal inflammation and impairs the anti-inflammatory effect of TGF-β1 in response to IL-1β triggered inflammatory response in vitro --- p.169 / Chapter 5.4.4.1 --- Conditional knockout of Smad4 increases the inflammatory cells infiltration --- p.169 / Chapter 5.4.4.2 --- Conditional knockout of Smad4 increases the TNFα expression in the UUO nephropathy --- p.171 / Chapter 5.4.4.3 --- Conditional knockout of Smad4 increases the IL-1β expression in the UUO nephropathy --- p.172 / Chapter 5.4.4.4 --- Conditional knockout of Smad4 increases the MCP-1 expression in the UUO nephropathy --- p.173 / Chapter 5.4.4.5 --- Conditional knockout of Smad4 increases the ICAM-1 level in the UUO nephropathy --- p.174 / Chapter 5.4.4.6 --- Time and dosage dependent experiments in response to IL-1β in macrophages --- p.175 / Chapter 5.4.4.7 --- Disruption of Smad4 in macrophages decreases the anti-inflammatory effect of TGF-β1 in response to IL-1β --- p.176 / Chapter 5.4.5 --- Disruption of Smad4 impairs the inhibitory effect of Smad7 on NFκB activation in vivo and in vitro --- p.178 / Chapter 5.4.5.1 --- Conditional knockout of Smad4 largely inhibits Smad7 level in UUO kidney --- p.178 / Chapter 5.4.5.2 --- Conditional knockout of Smad4 suppresses IκBα and further increases NF-κB p65 activation in UUO kidney --- p.180 / Chapter 5.4.5.3 --- Disruption of Smad4 inhibits Smad7 synthesis in macrophages --- p.182 / Chapter 5.4.5.4 --- Conditional knockout of Smad4 impair the inhibition effect of TGF-β1 on the activation of NFκB p65 in macrophages --- p.184 / Chapter 5.5 --- DISCUSSION --- p.186 / Chapter 5.6 --- CONCLUSION --- p.189 / Chapter CHAPTER VI --- SUMMARY AND DISCUSSION OF THE MAJOR FINDINGS --- p.190 / Chapter 6.1 --- SUMMARY AND DISCUSSION --- p.192 / Chapter 6.1.1 --- The diverse role of TβRII in renal inflammation and fibrosis both in vivo and in vitro --- p.192 / Chapter 6.1.2 --- Smad2 protects renal fibrosis by counter-regulating Smad3 signaling --- p.192 / Chapter 6.1.3 --- Disruption of Smad4 increased renal inflammation while suppressed the renal fibrosis in vivo and in vitro --- p.194 / Chapter 6.1.4 --- Comparative analysis of functions and related mechanisms between TβRII and Smad4 in renal disease --- p.195 / Chapter 6.1.5 --- Inadequacies of current work and future plan --- p.197 / Chapter 6.1.6 --- Perspectives (1) : The balance within the TGF-b/Smad signaling may determine the fate of renal diseases --- p.197 / Chapter 6.1.7 --- Perspectives(2)：The balance within the TGF-β/Smad signaling may determine the fate of renal diseases --- p.198 / Chapter 6.2 --- CONCLUSION --- p.201 / REFERENCES --- p.202 / PUBLICATION LIST --- p.232 / HONORS AND AWARDS --- p.237

<p>Psykisk ohälsa ökar i samhället, 20 % av Sveriges befolkning riskerar att någon gång i livet drabbas av depression. 600 personer varje år drabbas av kronisk njursvikt vilket innebär att dialysbehandling kan bli aktuellt. Att leva med en livslång sjukdom, som det innebär när man behandlas med dialys, och samtidigt drabbas av psykisk ohälsa innebär ett stort lidande för den enskilda individen. Genom att sjuksköterskan bemöter patienterna med empati och medmänsklighet så skapar denne goda förutsättningar för att relationen ska präglas av tillit och ärlighet. Syftet med denna litteraturstudie var att belysa hur sjuksköterskan kan identifiera och förebygga depression hos människor som behandlas med dialys. Resultatet visar att depression är vanligt förekommande bland dialyspatienter. Depression hos dessa människor kan leda till sömnproblem, nutritionsproblem, känsla av förlust och social isolering. Identifiering av depression kan ske genom användning av bedömningsformulär, detta kan även användas som en åtgärd för att förebygga depression. Mer kvalitativ forskning inom ämnet hade varit ett bra sätt för att öka förståelsen för denna patientgrupps situation. Genom att öka kunskapen ökar också förståelsen för depression och vad det innebär att leva med en livslång sjukdom och denna komplexa situation.</p> / <p>Mental illness is increasing in society, 20% of Sweden's population is at risk to suffer some from depression at some point in their lives. 600 people each year suffer from chronic kidney failure, which can lead to the need for dialysis treatment. To live with a lifelong disease and suffer from mental illness at the same time exposes the individual for a great suffering. By responding to the patients with empathy and compassion the nurse creates good conditions for the relationship, to be able to be characterized by trust and honesty. The purpose of this literature study was to elucidate how the nurse can identify and prevent depression for people treated with dialysis. The results show that depression is common among dialysis patients. For these people, depression can lead to sleeping problems, nutritional problems, feelings of loss, and social isolation. Identification of depression can be done through the use of evaluation forms; this can also be used as a measure to prevent depression. More qualitative research on the subject would be a good method to increase the understanding of the situation these patients are in. Raising awareness also increases the understanding of depression and what it means to live with a lifelong disease, and the complex situation it means.</p>

Diabetic long-term complications, despite intensive treatment, cause serious handicaps at relatively young age in diabetic patients. Diabetic nephropathy (DN) develops in up to 30% of patients with type 1 diabetes (T1D). Besides the eventual loss of kidney function, with need for dialysis treatment and transplantation, this complication also increases the risk of early death from cardiovascular disease. In addition to hyperglycaemia, the risk of developing DN is influenced by a number of life-style related factors, such as smoking and diet, but the mechanisms of action of these factors are largely unknown. The incidence of DN is not linearly related to diabetes duration. There is a peak incidence of DN at 15-20 years and this, together with results from family studies, shows that genetic factors are important contributors. Possible candidate genes are those involved in regulation of intraglomerular pressure and blood pressure, oxidative stress and inflammation. The main aims of this thesis were: ● To investigate the risk of DN associated with polymorphisms in; A) the endothelial NO-synthase gene (NOS3) and genes in the renin-angiotensin-system (RAAS) (all involved in the regulation of intraglomerular pressure). B) the manganese superoxide dismutase gene (SOD2) (involved in the regulation of oxidative stress). C) the ICAM1 gene (involved in activation and migration of lymphocytes) ● To investigate gene-smoking interactions ● To investigate the influence of normal diet on risk of microalbuminuria. The aims were addressed in different case-control settings, including 347 T1D patients from Sweden and 1163 patients from Finland, with or without DN, defined as; overt DN – having albumin excretion rate (AER) ≥200 μg/min, incipient DN – AER between 20 and 200 μg/min, non-DN controls – having AER &lt;20 μg/min and at least 20 years of diabetes duration. In one study also non-diabetic healthy individuals were included to asses the risk of T1D associated with the ICAM1 gene. Results: The RAAS genes were investigated in the Swedish sample set and there was an association between a polymorphism in the angiotensin II type 1 receptor (AGTR1) gene and overt DN, when adjusting for age, duration of diabetes, HbA1c, sex and smoking (adjusted OR=3.04, 99% CI=1.02-9.06). Also a synergistic interaction with smoking was indicated. The ICAM1 gene was investigated in the Swedish sample set, but no association with DN was found. There were, however, associations between T1D and two polymorphisms in this gene, rs281432 (OR=1.64, 95% CI=1.14-2.38) and rs5498 (OR=2.46, 95% CI=1.59-3.80). In the combined Swedish/Finnish sample set, the Glu/Glu genotype of the Glu298Asp polymorphism in the NOS3 gene was associated with DN when age at diabetes onset, duration of diabetes, HbA1c, blood pressure, sex and smoking were taken into account (adjusted OR=1.46, 95% CI=1.12-1.91). There was also association between a polymorphism in the MnSOD gene and DN in this sample set. Homozygosity for the valine-allele of the Val16Ala polymorphism was associated with increased risk of DN in a model including age at diabetes onset, duration of diabetes, HbA1c, sex and smoking (adjusted OR=1.32, 95% CI=1.00-1.74). Smoking was associated with DN (OR=2.00, 95% CI=1.60-2.50) and in the Swedish sample set there were indications of interactions between smoking and the NOS3 and SOD2 genes, but these results could not be confirmed in the Finnish sample set. A high protein intake can enhance glomerular filtration rate and accelerate progression to DN, also other dietary components such as fat, fibres, vitamins and the ratio red/white meat have been discussed as important for DN development. In a nested case-control study including young T1D patients, the normal dietary intakes of protein and other nutrients were assessed using a semiquantitative questionnaire. The results showed that T1D patients consuming more than 6.5 g fish protein (&gt;75th percentile) per day were at slightly lower risk to have microalbuminuria in both crude (OR=0.49, 95% CI=0.25-0.97) and adjusted analyses (OR=0.26, 95% CI=0.09-0.76, adjusted for age, duration of diabetes, sex, HbA1c, mean arterial pressure, BMI, region, smoking, energy intake and fish fat intake). Conclusions: The risk of having diabetic nephropathy is influenced by at least two genes controlling blood pressure and one gene protecting against oxidative stress. Smoking also increases the risk of DN and our findings indicate that smoking may accentuate the effect of the AGTR1, NOS3 and SOD2 genes. Normal dietary intake of protein was not associated with risk of having microalbuminuria in young T1D patients, on the other hand, an intake of fish protein above the 75th percentile decreased the risk of microalbuminuria.

Psykisk ohälsa ökar i samhället, 20 % av Sveriges befolkning riskerar att någon gång i livet drabbas av depression. 600 personer varje år drabbas av kronisk njursvikt vilket innebär att dialysbehandling kan bli aktuellt. Att leva med en livslång sjukdom, som det innebär när man behandlas med dialys, och samtidigt drabbas av psykisk ohälsa innebär ett stort lidande för den enskilda individen. Genom att sjuksköterskan bemöter patienterna med empati och medmänsklighet så skapar denne goda förutsättningar för att relationen ska präglas av tillit och ärlighet. Syftet med denna litteraturstudie var att belysa hur sjuksköterskan kan identifiera och förebygga depression hos människor som behandlas med dialys. Resultatet visar att depression är vanligt förekommande bland dialyspatienter. Depression hos dessa människor kan leda till sömnproblem, nutritionsproblem, känsla av förlust och social isolering. Identifiering av depression kan ske genom användning av bedömningsformulär, detta kan även användas som en åtgärd för att förebygga depression. Mer kvalitativ forskning inom ämnet hade varit ett bra sätt för att öka förståelsen för denna patientgrupps situation. Genom att öka kunskapen ökar också förståelsen för depression och vad det innebär att leva med en livslång sjukdom och denna komplexa situation. / Mental illness is increasing in society, 20% of Sweden's population is at risk to suffer some from depression at some point in their lives. 600 people each year suffer from chronic kidney failure, which can lead to the need for dialysis treatment. To live with a lifelong disease and suffer from mental illness at the same time exposes the individual for a great suffering. By responding to the patients with empathy and compassion the nurse creates good conditions for the relationship, to be able to be characterized by trust and honesty. The purpose of this literature study was to elucidate how the nurse can identify and prevent depression for people treated with dialysis. The results show that depression is common among dialysis patients. For these people, depression can lead to sleeping problems, nutritional problems, feelings of loss, and social isolation. Identification of depression can be done through the use of evaluation forms; this can also be used as a measure to prevent depression. More qualitative research on the subject would be a good method to increase the understanding of the situation these patients are in. Raising awareness also increases the understanding of depression and what it means to live with a lifelong disease, and the complex situation it means.

Thesis (Masters) -- The College of Saint Elizabeth, 2009. / Typescript. Available at The College of Saint Elizabeth - Office of Graduate Programs. "December 2009"

Introdução: Nefropatia induzida por contraste (NIC) está associada a piores desfechos intra-hospitalar e a longo prazo. Estudos recentes sugerem que hidratação com bicarbonato de sódio possa ser útil na sua prevenção; no entanto, essa medida de prevenção de NIC não foi, ainda, avaliada em pacientes diabéticos. Método: Subanálise de um estudo multicêntrico envolvendo 301 pacientes com creatinina sérica ≥ 1,2 mg/dl ou depuração de creatinina endógena (DCE) < 50 ml/min submetidos a cineangiocoronariografia ou angioplastia coronária, randomizados para receber hidratação com bicarbonato de sódio ou solução salina (soro fisiológico – SF) a 0,9%. Todos os procedimentos foram realizados com contraste iônico de baixa osmolaridade. Os desfechos avaliados foram incidência de NIC (definida como aumento de 0,5 mg/dl) e variação da creatinina e da DCE em 48 horas após o procedimento. Da totalidade de pacientes, 87 tinham diabetes melito e foram incluídos nesse subestudo. Resultados: Não houve diferença entre os grupos em relação a características demográficas, volume de contraste e níveis basais de creatinina e DCE. Entre os pacientes avaliados, 8 desenvolveram NIC: 4 (9,8%) pacientes no grupo bicarbonato e 4 (8,9%) no grupo SF 0,9% (p = 0,9). A variação da creatinina sérica e da DCE foi similar entre os grupos. Conclusão: Hidratação com bicarbonato de sódio não demonstrou benefício em reduzir a incidência de NIC em pacientes diabéticos submetidos a cateterismo cardíaco ou angioplastia coronária em comparação à hidratação com SF 0,9%. / Background: Contrast-induced nephropathy (CIN) is associated with worse clinical outcomes both at short and long-term follow-up. Recent evidence indicates that intravenous hydration with sodium-bicarbonate may reduce the incidence of CIN. However, this strategy has not been reported in diabetic patients. Methods: Sub-analysis of a multicenter study involving 301 patients with serum creatinine ≥ 1,2 mg/dL or creatinine clearance < 50 mL/min submitted to coronary angiography or percutaneous coronary intervention and randomized to intravenous hydration with sodium-bicarbonate or normal saline. All patients received low-osmolar contrast media. We assessed the incidence of CIN (defined as creatinine increase ≥ 0,5 mg/dL), and the average change in creatinine and creatinine clearance 48 hours after the procedure. A total of 87 diabetic patients were analyzed. Results: There was no difference between groups regarding baseline characteristics, contrast volume used, baseline creatinine levels and creatinine clearance. Eight patients presented CIN: 4 (9.8%) in the bicarbonate group and 4 (8.9%) in the saline group (p = 0.9). The average change in serum creatinine and creatinine clearance were similar between groups. Conclusion: Intravenous hydration with sodium-bicarbonate did not reduce the incidence of CIN in diabetic patients undergoing coronary angiography or percutaneous coronary intervention as compared to hydration with normal saline.

Introdução: Nefropatia induzida por contraste (NIC) está associada a piores desfechos intra-hospitalar e a longo prazo. Estudos recentes sugerem que hidratação com bicarbonato de sódio possa ser útil na sua prevenção; no entanto, essa medida de prevenção de NIC não foi, ainda, avaliada em pacientes diabéticos. Método: Subanálise de um estudo multicêntrico envolvendo 301 pacientes com creatinina sérica ≥ 1,2 mg/dl ou depuração de creatinina endógena (DCE) < 50 ml/min submetidos a cineangiocoronariografia ou angioplastia coronária, randomizados para receber hidratação com bicarbonato de sódio ou solução salina (soro fisiológico – SF) a 0,9%. Todos os procedimentos foram realizados com contraste iônico de baixa osmolaridade. Os desfechos avaliados foram incidência de NIC (definida como aumento de 0,5 mg/dl) e variação da creatinina e da DCE em 48 horas após o procedimento. Da totalidade de pacientes, 87 tinham diabetes melito e foram incluídos nesse subestudo. Resultados: Não houve diferença entre os grupos em relação a características demográficas, volume de contraste e níveis basais de creatinina e DCE. Entre os pacientes avaliados, 8 desenvolveram NIC: 4 (9,8%) pacientes no grupo bicarbonato e 4 (8,9%) no grupo SF 0,9% (p = 0,9). A variação da creatinina sérica e da DCE foi similar entre os grupos. Conclusão: Hidratação com bicarbonato de sódio não demonstrou benefício em reduzir a incidência de NIC em pacientes diabéticos submetidos a cateterismo cardíaco ou angioplastia coronária em comparação à hidratação com SF 0,9%. / Background: Contrast-induced nephropathy (CIN) is associated with worse clinical outcomes both at short and long-term follow-up. Recent evidence indicates that intravenous hydration with sodium-bicarbonate may reduce the incidence of CIN. However, this strategy has not been reported in diabetic patients. Methods: Sub-analysis of a multicenter study involving 301 patients with serum creatinine ≥ 1,2 mg/dL or creatinine clearance < 50 mL/min submitted to coronary angiography or percutaneous coronary intervention and randomized to intravenous hydration with sodium-bicarbonate or normal saline. All patients received low-osmolar contrast media. We assessed the incidence of CIN (defined as creatinine increase ≥ 0,5 mg/dL), and the average change in creatinine and creatinine clearance 48 hours after the procedure. A total of 87 diabetic patients were analyzed. Results: There was no difference between groups regarding baseline characteristics, contrast volume used, baseline creatinine levels and creatinine clearance. Eight patients presented CIN: 4 (9.8%) in the bicarbonate group and 4 (8.9%) in the saline group (p = 0.9). The average change in serum creatinine and creatinine clearance were similar between groups. Conclusion: Intravenous hydration with sodium-bicarbonate did not reduce the incidence of CIN in diabetic patients undergoing coronary angiography or percutaneous coronary intervention as compared to hydration with normal saline.

Introdução: Nefropatia induzida por contraste (NIC) está associada a piores desfechos intra-hospitalar e a longo prazo. Estudos recentes sugerem que hidratação com bicarbonato de sódio possa ser útil na sua prevenção; no entanto, essa medida de prevenção de NIC não foi, ainda, avaliada em pacientes diabéticos. Método: Subanálise de um estudo multicêntrico envolvendo 301 pacientes com creatinina sérica ≥ 1,2 mg/dl ou depuração de creatinina endógena (DCE) < 50 ml/min submetidos a cineangiocoronariografia ou angioplastia coronária, randomizados para receber hidratação com bicarbonato de sódio ou solução salina (soro fisiológico – SF) a 0,9%. Todos os procedimentos foram realizados com contraste iônico de baixa osmolaridade. Os desfechos avaliados foram incidência de NIC (definida como aumento de 0,5 mg/dl) e variação da creatinina e da DCE em 48 horas após o procedimento. Da totalidade de pacientes, 87 tinham diabetes melito e foram incluídos nesse subestudo. Resultados: Não houve diferença entre os grupos em relação a características demográficas, volume de contraste e níveis basais de creatinina e DCE. Entre os pacientes avaliados, 8 desenvolveram NIC: 4 (9,8%) pacientes no grupo bicarbonato e 4 (8,9%) no grupo SF 0,9% (p = 0,9). A variação da creatinina sérica e da DCE foi similar entre os grupos. Conclusão: Hidratação com bicarbonato de sódio não demonstrou benefício em reduzir a incidência de NIC em pacientes diabéticos submetidos a cateterismo cardíaco ou angioplastia coronária em comparação à hidratação com SF 0,9%. / Background: Contrast-induced nephropathy (CIN) is associated with worse clinical outcomes both at short and long-term follow-up. Recent evidence indicates that intravenous hydration with sodium-bicarbonate may reduce the incidence of CIN. However, this strategy has not been reported in diabetic patients. Methods: Sub-analysis of a multicenter study involving 301 patients with serum creatinine ≥ 1,2 mg/dL or creatinine clearance < 50 mL/min submitted to coronary angiography or percutaneous coronary intervention and randomized to intravenous hydration with sodium-bicarbonate or normal saline. All patients received low-osmolar contrast media. We assessed the incidence of CIN (defined as creatinine increase ≥ 0,5 mg/dL), and the average change in creatinine and creatinine clearance 48 hours after the procedure. A total of 87 diabetic patients were analyzed. Results: There was no difference between groups regarding baseline characteristics, contrast volume used, baseline creatinine levels and creatinine clearance. Eight patients presented CIN: 4 (9.8%) in the bicarbonate group and 4 (8.9%) in the saline group (p = 0.9). The average change in serum creatinine and creatinine clearance were similar between groups. Conclusion: Intravenous hydration with sodium-bicarbonate did not reduce the incidence of CIN in diabetic patients undergoing coronary angiography or percutaneous coronary intervention as compared to hydration with normal saline.

A doença renal policística autossômica dominante (DRPAD) constitui-se na enfermidade humana monogênica com risco de óbito mais frequente, responsabilizando-se por 4,4 a 10,0% dos casos de doença renal terminal em diferentes populações. Na quase totalidade dos pacientes, a doença é causada por mutação em um de dois genes: PKD1 (polycystic kidney disease 1) ou PKD2 (polycystic kidney disease 2). Tais genes codificam, respectivamente, as proteínas policistina-1 (PC1) e policistina-2 (PC2). Mutações em PKD1, por sua vez, respondem pela ampla maioria dos casos de DRPAD. Camundongos haploinsuficientes para Pkd1 (Pkd1+/-), o gene ortólogo a PKD1 neste animal, consistem num modelo não cístico de deficiência de atividade deste gene. Em um estudo anterior, mostramos que animais Pkd1+/- apresentam lesão renal mais severa que camundongos selvagens (Pkd1+/+) quando submetidos a isquemia/reperfusão (I/R) renal. Esse estudo sugeriu, portanto, que a capacidade de regeneração renal pós-I/R esteja prejudicada em camundongos Pkd1+/- e em pacientes com DRPAD. O insulto por I/R constitui-se em uma causa importante de indução de estresse de retículo endoplasmático (ER), podendo ativar as vias UPR (unfolded protein response) e ERAD (ER-associated degradation). Além disso, a ativação de vias envolvidas no ER determinado por I/R exerce um efeito de agravamento da lesão decorrente deste insulto. O ER pode, ainda, ativar e ser induzido pela resposta inflamatória. Estudos prévios revelaram que as policistinas também se relacionam com este processo. A expressão de PC2 pode ser superregulada pela ativação de um dos braços da via UPR, enquanto a ativação da via ERAD estimula sua degradação. A superexpressão de XBP1, por sua vez, atenua o fenótipo cístico em camundongos deficientes em Pkd1, revelando que a ativação da via UPR pode mitigar a formação cística. Para analisar a relação entre ER e suscetibilidade aumentada a I/R na deficiência de Pkd1, avaliamos diferentes marcadores de ER em camundongos Pkd1+/- e Pkd1+/+ submetidos a um insulto leve por I/R renal associado a 32 min de isquemia. A razão de expressão renal dos mRNAs Xbp1s/Xbp1u mostrou-se menor em camundongos Pkd1+/- que Pkd1 +/+ 48 h após I/R, enquanto a expressão proteica de XBP1s foi maior em rins Pkd1+/- comparados a Pkd1+/+ após o insulto. Não detectamos diferença na expressão renal do gene Hspa5 e de seu produto BIP/GRP78, assim como na expressão de Ddit3, gene que codifica CHOP, após intervenção sham e após I/R. Também não observamos diferenças entre os níveis renais e séricos de IL1beta, IL6, IL10, TNFalfa e RANTES entre camundongos Pkd1+/- e Pkd1+/+ pós-procedimento sham e pós-I/R, embora tendências não significantes de elevação de MCP1 tenham sido detectadas nos rins submetidos ao insulto para ambos os genótipos. As variações em sentidos opostos de XBP1s e Xbp1s/Xbp1u determinadas por I/R em rins Pkd1+/- são consistentes com uma maior suscetibilidade destes animais à indução de ER. Esses achados sugerem que a indução de ER em resposta a um insulto leve por I/R possa aumentar a atividade de PC1 e exercer um efeito de atenuação sobre a maior suscetibilidade de camundongos deficientes em Pkd1 a I/R renal / Autosomal dominant polycystic kidney disease (ADPKD) is the most common life-threatening monogenic disease in humans, accounting for 4.4 to 10.0% of the end-stage kidney disease cases in different populations. In almost all patients, this disorder is caused by a mutation in one of two genes: PKD1 (polycystic kidney disease 1) or PKD2 (polycystic kidney disease 2). These genes encode, respectively, the proteins polycystin-1 (PC1) and polycystin-2 (PC2). Mutations in PKD1, in turn, are responsible for the large majority of ADPKD cases. Pkd1- haploinsufficient mice (Pkd1+/-), the gene orthologous to PKD1 in this animal, constitute a noncystic model of this gene\'s deficiency. In a previous study, we showed that Pkd1+/- animals develop a more severe renal injury than wild-type mice (Pkd1+/+) when submitted to renal ischemia/reperfusion (I/R). This study suggested, therefore, that the capacity of renal regeneration following I/R is impaired in Pkd1+/- mice and in ADPKD patients. The I/R insult is an important cause of endoplasmic reticulum stress (RS) induction, potentially leading to activation of the UPR (unfolded protein response) and ERAD (ER-associated degradation) pathways. The activation of pathways involved in RS determined by I/R exerts an aggravating effect on the injury resulting from the insult. In addition, RS can activate and be induced by the inflammatory response. Previous studies revealed that polycystins also relate to this process. PC2 expression can be upregulated by the activation of one of the UPR pathway branches, while activation of the ERAD pathway stimulates its degradation. XBP1 overexpression, in turn, attenuates the cystic phenotype in Pkd1-deficient mice, revealing that activation of UPR can mitigate cyst formation. To analyze the relationship between RS and the increased susceptibility to I/R associated to Pkd1 deficiency, we evaluated different RS markers in Pkd1+/- and Pkd1+/+ mice submitted to a mild I/R insult determined by 32-min ischemia. The renal expression ratio of mRNA Xbp1s/Xbp1u was lower in Pkd1+/- than Pkd1+/+ mice 48 h after I/R, while the XBP1s protein expression was higher in Pkd1+/- compared to Pkd1+/+ kidneys after the insult. We have not detected differences in renal expression of the Hspa5 gene and its product BIP/GRP78, as well as in Ddit3 expression, the gene that encodes CHOP, postsham intervention and post-I/R. We have also not observed differences in the renal and serum levels of IL1beta, IL6, IL10, TNFalfa and RANTES between Pkd1+/- e Pkd1+/+ mice post-sham procedure and post-I/R, although non-significant trends of MCP1 increase have been detected in kidneys submitted to the insult for both genotypes. The variations in different directions of XBP1s and Xbp1s/Xbp1u induced by I/R in Pkd1+/- kidneys are consistent with a higher susceptibility of these animals to RS induction. These findings suggest that the RS induction in response to a mild I/R insult can increase PC1 activity and exert an attenuating effect on the increased susceptibility of Pkd1-deficient mice to renal I/R

Background: Low protein diet is one of the treatments for patients with chronic kidney disease during the pre-dialysis period. Studies about this subject have been published for more than 20 years, but the conclusions about its effect and the most appropriate composition are not clear. A low-protein diet is recommended for some patients at the Nephrology Clinic of General University Hospital. These patients are educated and re- educated by a nutrition therapist who also controls their food intake through a food diary. Objectives: To evaluate the effect of low-protein diet on the progression of chronic kidney disease in our conditions and to compare the results with the control group. Furthermore, to evaluate the nutritional status of these patients and the influence of the diet on it. Last but not least, describe a set of patients coming to education about low-protein diet. Methods: The input data was retrospectively retrieved from the medical records of patients who attended medical checks at the Nephrology Clinic of General University Hospital between 10/2016 and 3/2018. A group of 15 patients with a low-protein diet was compared with a control group of 15 patients who did not receive low-protein diet education. The main data collected was year of birth, estimated GFR, urea, creatinine, albumin,...