Similarly, inhibitor order us Hcy induced p85 PI3K phosphorylation in a time dependent manner. Phosphorylation of p85 PI 3K significantly increased at 20 minutes Hcy pared with levels at the initiation of the study. At 30 min utes, p85 PI 3K phosphorylation decreased as compared with 20 minutes. Hcy induced p38MAPK andadhesion to mesangial cellsand by MIP 2 Modulates Leukocyte cell adhesion to mesangial cells Hcy induced leukocyte adhesion to MC was determined by cell adhesion assay following incubation of with Hcy. L Cys represented control condition. L Cys did not have a significant effect on leukocyte adhesion to MC whereas Hcy induced dose dependent increase in leukocyte adhesion to mesangial cells. Leuko cyte adhesion increased significantly up to 1. 8 fold at 50 M Hcy compared with control condition.

SB203580 and LY294002 treated MC was employed to determine the role of p38MAPK and PI 3K in MIP 2 medi ated leukocyte adhesion to these glomerular cells. As revealed, LY294002 and SB203580 blocked leukocyte adhesion induced by 50 M Hcy. Blocking anti body against MIP 2 confirmed the functional role of MIP 2 in Hcy induced leukocyte adhesion to MC. Hcy induced leukocyte adhesion to MC was sig nificantly blocked up to 3 fold by MIP 2 antibody. Discussion MIP 2 is a C C chemokine, known to recruit neu trophils and studies suggest that neutrophil recruit ment may bear relevance to the development and progression of glomerular diseases. The initial indication that MIP 2 may participate in glomerular disease arose from observations that isolated glomeruli and MC pro duced MIP 2 in response to immune comple es.

Sub sequently, in another in vivo rat model of mesangioproliferative glomerulonephritis , glomerular nitric o ide was shown to be capable of inducing MIP 2 e pression, which in turn lead to neu trophil recruitment. Kidney disease is associated with increases in plasma Hcy and Hcy induces MCP 1 pro duction by glomerular MC. In order to identify cytokines whose e pression may be increased by Hcy, we initially employed antibody array approach to evaluate cytokine production by MC e posed to pathophysiologic levels of Hcy. Our initial observation was that elevated e tra cellular Hcy increased the levels of cytokines, TIMP 1 and MIP 2. For another cytokine, MCP 1 there was a 20 percent increase in protein levels, but this was not statistically significant.

Other Dacomitinib studies have dem onstrated a 20 to 40 percent increase in MCP 1 by MC and hepatocytes e posed to comparable concentra tions of Hcy. Hence, our observations are similar to the aforementioned reports, but in the current study, Hcy induced MCP 1 changes were not significant. In contrast, the observations for TIMP 1 are consistent with earlier studies, while data relating to induction of MIP 2 by Hcy have not been previously reported.