Inflammation, Endothelial Dysfunction and Increased Left Ventricular Mass in Chronic Kidney Disease (CKD) Patients: A Longitudinal Study

<div><p>Introduction</p><p>Within this longitudinal study we investigated the association of inflammation markers C-reactive protein (CRP), interleukin-6 (IL-6) and tumor necrosis factor-α (TNFα) and endothelial dysfunction markers intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) with left ventricular mass indexed for height²·⁷¹ (LVMI) in hypertensive predialysis CKD patients.</p><p>Material and Methods</p><p>From 2004 to 2005, 182 incident consecutive adult patients from the outpatient CKD clinics of two hospitals in Greece with CKD and hypertension or using antihypertensive medication, were included. Of these, 107 patients underwent CRP (mg/l) and LVMI (g/height²·⁷¹) measurements annually for three years.</p><p>Results</p><p>In the longitudinal analyses, using linear mixed modeling, a higher IL-6 (ß = 1.9 (95%ci:0.38;3.5), inflammation score based on CRP, IL-6 and TNF-α (ß = 5.0 (95%ci:0.72; 9.4) and VCAM-1 (ß = 0.01 (95%ci:0.005;0.02) were associated with higher LVMI. These models were adjusted for age, gender and primary renal disease, and for confounders that on top changed the beta with ≥10%, i.e. diuretic use (for IL-6 and inflammation score).</p><p>Conclusion</p><p>The results suggest that in predialysis CKD patients, inflammation as well as endothelial dysfunction may play an important role towards the increase in LVMI.</p></div

Correlation between log TNF-α with left ventricular mass (g/m²·⁷¹) (n = 182).

<p>Correlation between log TNF-α with left ventricular mass (g/m²·⁷¹) (n = 182).</p

Correlation between VCAM-1 with left ventricular mass (g/m²·⁷¹) (n = 182).

<p>Correlation between VCAM-1 with left ventricular mass (g/m²·⁷¹) (n = 182).</p

The association between both inflammation markers and endothelial dysfunction markers with left ventricular mass using linear regression analysis in cross sectional data.

<p>The association between both inflammation markers and endothelial dysfunction markers with left ventricular mass using linear regression analysis in cross sectional data.</p

The association between inflammation markers and endothelial dysfunction markers and left ventricular mass per year using linear mixed modelling in longitudinal data.

<p>The association between inflammation markers and endothelial dysfunction markers and left ventricular mass per year using linear mixed modelling in longitudinal data.</p

Correlation between IL-6 with left ventricular mass (g/m²·⁷¹) (n = 182).

<p>Correlation between IL-6 with left ventricular mass (g/m²·⁷¹) (n = 182).</p

Mutant <i>COL4A3</i> chains expressed in AB8/13 cultured podocytes demonstrate a trend for increased intracellular retention.

<p>(a) AB8/13 cells were transiently transfected with expression vectors containing wild-type <i>COL4A3</i>-WT or the mutant <i>COL4A3</i> (p.G1334E, p.G871C, p.G484R, p.A587G) cDNAs that included a HA epitope at C-terminus. Single chain expression was measured via Western blot analysis of the cell lysate, 48 h after transfection. No HA antigen was detected in AB8/13 cells transfected with a construct expressing the empty vectors. Shown is a representative Western blot of proteins in cell lysates. (b) All mutant chains show a trend towards increased intracellular retention as compared to the wild type chain, although not reaching significance at the 48 h time point. Shown is densitometry analysis data normalized to tubulin expression. Data are represented as means ± SEM of n≥3 independent experiments.</p

Overexpression of wild type or mutant <i>COL4A3</i> chains induces XBP1 splicing in AB8/13 cells.

<p>(a) Representative experiment of reverse transcription-PCR using XBP1 mRNA as template, from AB8/13 cells transiently expressing <i>COL4A3</i>-WT (A3/WT) or the mutant chains G1334E, G871C, G484R (<i>COL4A3</i>). PCR products were run on 3% agarose gel. It is apparent that over-expression of all chains induces XBP1 splicing, as evidenced by the appearance of the spliced band (s) when the PCR product is cut with the restriction enzyme Pst1. (h) hybrid, (u) unspliced (b) RT-PCR is quantified via densitometric analysis of the bands after PstI digestion as follows: ratio of the spliced band and the sum of the two PstI digest bands (s/(u1+u2), with PstI digestion. Hybrid band (h) was considered as equally contributing to unspliced and spliced bands. There is statistically significant XBP1 splicing when either WT or any of the mutant chains is overexpressed in AB8/13 cells. L19 was used as an internal PCR control. Data are means ± SEM of three independent experiments.</p

Summary of pathogenic <i>COL4A3/A4</i> mutations found in Greek-Cypriot families studied here.

<p>One additional deletion mutation was detected in a Cypriot of Romanian origin.</p><p>Summary of pathogenic <i>COL4A3/A4</i> mutations found in Greek-Cypriot families studied here.</p

Τable 1. Information on mutations and reagents used for their identification.

<p>If no restriction enzyme is given, detection was performed by direct Sanger DNA sequencing.</p><p>Τable 1. Information on mutations and reagents used for their identification.</p