Brabykinin B1 Receptor Antagonism Is Beneficial in Renal Ischemia-Reperfusion Injury

Previously we have demonstrated that bradykinin B1 receptor deficient mice (B1KO) were protected against renal ischemia and reperfusion injury (IRI). Here, we aimed to analyze the effect of B1 antagonism on renal IRI and to study whether B1R knockout or antagonism could modulate the renal expression of pro and anti-inflammatory molecules. To this end, mice were subjected to 45 minutes ischemia and reperfused at 4, 24, 48 and 120 hours. Wild-type mice were treated intra-peritoneally with antagonists of either B1 (R-954, 200 µg/kg) or B2 receptor (HOE140, 200 µg/kg) 30 minutes prior to ischemia. Blood samples were collected to ascertain serum creatinine level, and kidneys were harvested for gene transcript analyses by real-time PCR. Herein, B1R antagonism (R-954) was able to decrease serum creatinine levels, whereas B2R antagonism had no effect. The protection seen under B1R deletion or antagonism was associated with an increased expression of GATA-3, IL-4 and IL-10 and a decreased T-bet and IL-1β transcription. Moreover, treatment with R-954 resulted in lower MCP-1, and higher HO-1 expression. Our results demonstrated that bradykinin B1R antagonism is beneficial in renal IRI

Correction: Bradykinin B1 Receptor Antagonism Is Beneficial in Renal Ischemia-Reperfusion Injury

Het doel van dit proefschrift was om te komen tot een plaatsbepaling van de calcium antagonist diltiazem bij de behandeling van patiënten met stabiele angineuze klachten en andere cardiale ischaemische syndromen. Diltiazem kwam 6 jaar geleden op de Nederlandse markt: een late (re)-evaluatie van dit antiangineuze middel leek nuttig om de volgende redenen. ... Zie: Samenvatting

Early modulation of inflammation by mesenchymal stem cell after acute kidney injury

Therapy with stem cells has showed to be promising for acute kidney injury (AKI), although how it works is still controversial. Modulation of the inflammatory response is one possible mechanism. Most of published data relies on early time and whether the protection is still maintained after that is not known. Here, we analyzed whether immune modulation continues after 24 h of reperfusion. MSC were obtained from male Wistar rats. After 3-5 passages, cells were screened for CD73, CD90, CD44, CD45, CD29 and CD 31. In addition, MSC were submitted to differentiation in adipocyte and in osteocyte. AKI was induced by bilaterally clamping of renal pedicles for 60 min. Six hours after injury, MSC (2 x 105 cells) were administered intravenously. MSC-treated animals presented the lowest serum creatinine compared to non-treated animals (24 h: 1.3 +/- 0.21 vs. 3.23 +/- 0.89 mg/dl, p<0.05). The improvement in renal function was followed by a lower expression of IL-1b, IL-6 and TNF-alpha and higher expression of IL-4 and IL-10. However, 48 h after reperfusion, this cytokine profile has changed. The decrease in Th1 cytokines was less evident and IL-6 was markedly up regulated. PCNA analysis showed that regeneration occurs faster in kidney tissues of MSC-treated animals than in controls at 24 h. And also ratio of Bcl-2/Bad was higher at treated animals after 24 and 48 h. Our data demonstrated that the immunomodulatory effects of MSC occur at very early time point, changing the inflammation profile toward a Th2 profile. (C) 2009 Elsevier B.V. All rights reserved

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Pro and anti-inflammatory molecule expression in B1KO and wild type animals.

<p>All molecule expressions were measured by real-time PCR at 24 hours of reperfusion. B1KO group had lower pro-inflammatory molecule expression (T-bet and IL-1β) (A and B) and higher anti-inflammatory response (GATA-3, IL-4 and IL-10) (C, D and E). Statistical analyses were performed using the t-test.* B1KO <i>versus</i> B1B2WT, p<0.05.</p

Renal IRI and bradykinin receptors expression.

<p>Bradykinin receptors were analyzed by real-time PCR. In B1B2WT, receptors expressions were cross-modulated (A). In B2KO, B1R expression was increased at 4 and 24 hours (B). Statistical analyses were performed by ANOVA. *B1R <i>versus</i> B2R, p<0.05. # B2KO <i>versus</i> B1B2WT, p<0.05.</p

Cell death modulation under B1R-knockout.

<p>Apoptosis was estimated by Bcl-2 and Bad expression and caspase-3 activity, at 24 hours of reperfusion. Bcl-2 and Bad expression were measured by real-time PCR, and caspase-3 activity by fluorimetric assay. B1KO animals presented higher Bcl-2 expression (A) along with lower Bad expression (B) and caspase-3 activity (C), indicating a lower degree of apoptosis. Statistical analyses were performed using ANOVA.* B1KO <i>versus</i> B1B2WT and B2KO, p<0.05.</p

Pro and anti-inflammatory molecules expression after B1R antagonist and agonist treatment.

<p>All molecule expressions were estimated by real-time PCR at 24 hours of reperfusion. B1R antagonism (R-954) resulted in lower pro-inflammatory molecule expression (T-bet, IL-1β and MCP-1) (A, B and C) and higher anti-inflammatory response (GATA-3, IL-4, IL-10 and HO-1) (D, E, F and G). Molecule expression after B1R agonism (DABK) were similar to non-treated mice. Statistical analyses were performed using ANOVA.*IR+HOE-140 <i>versus</i> IR, p<0.05.</p