DOI: 10.5061/dryad.k0p2ngfhx
title: Gut alterations in a chronic kidney disease rat model with diet-induced vascular calcification

description: Intestinal disorders and vascular calcification (VC) are often associated
 with chronic kidney disease (CKD). While gut barrier alterations have been
 reported in CKD (abnormal intestinal permeability, bacterial overgrowth
 and inflammation), it is not clear if vascular calcification influences
 such alterations. Bidirectional relationships between VC and gut
 dysfunction could be mediated by increased inflammation and uremic toxin
 generation. We used the SNx-VC model of uremic vascular calcification
 (rats undergoing subtotal 5/6th nephrectomy and fed a pro-calcifying
 high-phosphate and vitamin D diet) to investigate gut alterations in vivo.
 We confirmed the presence of CKD and VC (von Kossa), and observed
 increased gut-origin uremic toxin, indoxyl sulfate (IS), in SNx-VC animals
 compared to controls. In SNx-VC, we observed a decreased mucus production
 (Alcian blue, Mucin 2 staining) in colon and ileum that correlated with
 calcification severity. There was no change in inflammation markers or
 tight junction proteins expression. We assessed intestinal levels in the
 NOD‐like receptor family pyrin domain containing 6 (NLRP6) protein, known
 to regulate mucus secretion, and found no change in colon or ileum. NLRP6
 mRNA was however decreased in SNx-VC in colon, along with other mRNA
 (Ly96, Sod1), while Tlr2 was increased. Our observations of low mucus, low
 Nlrp6 mRNA, and high IS in SNx-VC confirm a link between gut barrier
 alterations and uremic VC. This suggests that alterations in the mucus
 layer could favour the generation of gut-origin uremic toxins and promote
 VC in CKD.
description: # Gut alterations in a chronic kidney disease rat model with diet-induced
 vascular calcification ### DOI Links This dataset contains all the data
 analysed in the related manuscript. For methods and statistical analyses,
 please refer to the manuscript. * The dataset:
 [https://doi.org/10.5061/dryad.k0p2ngfhx](https://doi.org/10.5061/dryad.k0p2ngfhx) * The article: [https://doi.org/10.1002/2211-5463.70043](https://doi.org/10.1002/2211-5463.70043) ## Description of the data ### File * Gut_alterations_in_SNXVC_rat.csv ### Structure * Each line represents one observational unit (one animal). * The columns refer to the experimental groups and individual values observed at the end of protocol. * Individual values correspond to blood, urine, kidney, aorta, colon, and ileum analysis (biological assays, histology, RTqPCR). * Values are reported in original scale and, if applicable, after Transformation. * Missing values are marked as "." ### Variables | Name | Unit | Explanation/Method | | :---------------------------- | :---------------------------- | :----------------------------------------------------------- | | rat\_id | n/a | Unique rat identifier | | group | n/a | Experimental group | | snx | 1=Yes=SNx; 0=No=Sham | SNx surgery | | VC\_diet | 1=yes; 0=No | Vascular Calcification (VC)-inducing diet | | VD\_dose | µg/kg/day | Vitamin D dose (given 5 days a week) in the VC-inducing diet | | calcium\_content\_aorta | nmol/100mg of aorta | O-cresolphthalein complexone assay | | calcium\_content\_aorta\_rk | rank | Transformation | | aorta\_vkossa | positive tissue area in % | Von Kossa staining | | Creatininemia\_micM | µM | Blood test | | Creatininemia\_micM\_log | ln(µM) | Transformation | | Kidney\_siriusred | positive tissue area in % | Sirius Red staining | | Kidney\_siriusred\_log | ln(positive tissue area in %) | Transformation | | Uremia\_mM | mM | Blood test | | Uremia\_mM\_log | ln(mM) | Transformation | | Phosphatemia\_mM | mM | Blood test | | Phosphatemia\_mM\_rk | rank | Transformation | | PxCa\_mM2 | mM² | Blood test | | PxCa\_mM2\_log | ln(mM²) | Transformation | | calcemia\_mM | mM | Blood test | | proteinuria\_g\_L | g/L | Urine test | | proteinuria\_g\_L\_log | ln(mM) | Transformation | | indoxylsulfate\_plasma | µg/L | HPLC | | indoxylsulfate\_plasma\_log | ln(µg/L) | Transformation | | pcresylsulfate\_plasma | µg/L | HPLC | | pcresylsulfate\_plasma\_log | ln(µg/L) | Transformation | | mucus\_alcian\_colon | positive tissue area in % | Alcian Blue staining | | mucus\_alcian\_ileum | positive tissue area in % | Alcian Blue staining | | mucus\_muc2\_colon | positive tissue area in % | Alcian Blue staining | | mucus\_muc2\_ileum | positive tissue area in % | Alcian Blue staining | | mucus\_muc2\_ileum\_log | ln(positive tissue area in %) | Transformation | | inflammation\_HE\_colon | score | Hematoxylin Eosin staining | | inflammation\_HE\_ileum | score | Hematoxylin Eosin staining | | inflammation\_HE\_ileum\_log | ln(score) | Transformation | | macrophages\_colon | positive area in % | CD68 immunostaining | | macrophages\_ileum | positive area in % | CD68 immunostaining | | macrophages\_ileum\_log | ln(positive tissue area in %) | Transformation | | NLRP6\_staining\_colon | positive tissue area in % | NLRP6 immunostaining | | NLRP6\_staining\_colon\_rk | rank | Transformation | | NLRP6\_staining\_ileum | positive tissue area in % | NLRP6 immunostaining | | NLRP6\_WB\_colon | AU | Western Blot arbitrary unit | | NLRP6\_WB\_colon\_log | ln(AU) | Transformation | | claudin1\_WB\_colon | AU | Western Blot arbitrary unit | | claudin1\_WB\_colon\_log | ln(AU) | Transformation | | occludin\_WB\_colon | AU | Western Blot arbitrary unit | | aoah\_gene\_colon | mRNA expression ratio | RT-qPCR | | aoah\_gene\_colon\_scaled | ratio/mean control ratio | Transformation | | catalase\_gene\_colon | mRNA expression ratio | RT-qPCR | | catalase\_gene\_colon\_scaled | ratio/mean control ratio | Transformation | | claudin1\_gene\_colon | mRNA expression ratio | RT-qPCR | | claudin1\_gene\_colon\_scaled | ratio/mean control ratio | Transformation | | claudin2\_gene\_colon | mRNA expression ratio | RT-qPCR | | claudin2\_gene\_colon\_scaled | ratio/mean control ratio | Transformation | | claudin4\_gene\_colon | mRNA expression ratio | RT-qPCR | | claudin4\_gene\_colon\_scaled | ratio/mean control ratio | Transformation | | il10\_gene\_colon | mRNA expression ratio | RT-qPCR | | il10\_gene\_colon\_scaled | ratio/mean control ratio | Transformation | | il18\_gene\_colon | mRNA expression ratio | RT-qPCR | | il18\_gene\_colon\_scaled | ratio/mean control ratio | Transformation | | il6\_gene\_colon | mRNA expression ratio | RT-qPCR | | il6\_gene\_colon\_scaled | ratio/mean control ratio | Transformation | | mcp1\_gene\_colon | mRNA expression ratio | RT-qPCR | | mcp1\_gene\_colon\_scaled | ratio/mean control ratio | Transformation | | md2\_gene\_colon | mRNA expression ratio | RT-qPCR | | md2\_gene\_colon\_scaled | ratio/mean control ratio | Transformation | | mucin2\_gene\_colon | mRNA expression ratio | RT-qPCR | | mucin2\_gene\_colon\_scaled | ratio/mean control ratio | Transformation | | nfkb\_gene\_colon | mRNA expression ratio | RT-qPCR | | nfkb\_gene\_colon\_scaled | ratio/mean control ratio | Transformation | | nlrp6\_gene\_colon | mRNA expression ratio | RT-qPCR | | nlrp6\_gene\_colon\_scaled | ratio/mean control ratio | Transformation | | occludin\_gene\_colon | mRNA expression ratio | RT-qPCR | | occludin\_gene\_colon\_scaled | ratio/mean control ratio | Transformation | | sod1\_gene\_colon | mRNA expression ratio | RT-qPCR | | sod1\_gene\_colon\_scaledd | ratio/mean control ratio | Transformation | | tlr2\_gene\_colon | mRNA expression ratio | RT-qPCR | | tlr2\_gene\_colon\_scaled | ratio/mean control ratio | Transformation | | tlr4\_gene\_colon | mRNA expression ratio | RT-qPCR | | tlr4\_gene\_colon\_scaled | ratio/mean control ratio | Transformation | | zo1\_gene\_colon | mRNA expression ratio | RT-qPCR | | zo1\_gene\_colon\_scaled | ratio/mean control ratio | Transformation |