Ges ttigt. Ang II increased Ht the intracellular Ren levels with increasing concentrations of Ang II in the proteomic ZD4054 Zibotentan juice mixture appearance 0.7-fold with 10 M Ang II also increased Ht times and 0 Times in intracellular Ren levels of angiotensin II were observed with 10 M and 10 M Ang II listed. In separate experiments, mesangial cells were incubated with NG or NG with 10 M Ang II and the mixture for 15 minutes proteotypic juice 2h. A 1.5-fold erh Increase of Ang II levels in cell lysates was observed after 15 min incubation. In addition, intracellular Re Ang II levels increased Ht times for 30 min, times after 60 minutes, and Hours after 120 minutes. These results showed a Erh Increase the concentration and time dependent Ngig of the intracellular Ren levels of Ang II in response to angiotensin II in mesangial cells transfection.
3.3. Intracellular Re Ang II increased Ht TGF b1, collagen IV, fibronectin and cell proliferation. Then, the effects of increased Hten intracellular Ren levels of Ang II TGF-b1 and matrix proteins Such as collagen IV and fibronectin, the cells with 5 mM glucose determined.Mesangial NG were incubated alone or with a BIX 02189 mixture of Ang II and for proteomic juice 24 h the cells and media were analyzed for TGF b1, collagen IV, fibronectin, and layers. In cells transfected Ang II TGF b1 levels were significantly elevated ht control In comparison to cells. After 24 h of treatment, attempts have been set, and total cell lysates were prepared. Protein expression of total Stat3 was determined byWestern blotting.
As shown in Figure 4, mesangial cells were treated with Ang II or transfected with exogenous Ang II protein by a increased Stat3 hte expression of the transcription factor. Treated Densitometric analysis of Western blots showed a significant Erh Increase of Stat3 protein expression in cells with exogenous Ang II was blocked by candesartan. These observations are consistent with previous reports that activation of Stat3 by exogenous Ang II via AT1 receptor. In mesangial cells transfected with Ang II significantly increased Ht the expression of Stat3 protein was increased in response Hte intracellular Observed re Ang II. Since cells of NG t Ang II group were treated with candesartan before transfection with Ang II, these results suggest, speculated that the intracellular effects of angiotensin II on the Is re Stat3 protein by the AT 1 receptor-associated extracellular path notmediated signaling.
3.4.2. Stat3 phosphorylation. Mesangial cells were cultured in 96-well plates, washed with 5 mM glucose or NG, containing 1 M Ang II or mixed with proteotypic juice NG with 1 M exogenous Ang II for 20 minutes. at the end of the incubation period the cells were fixed and were analyzed for levels of total Stat3 pStat3 or cells treated as described inMethods.Mesangial with exogenous Ang II, showed a significant increase in phosphorylation of STAT3 in serine 727 and tyrosine residues at a time 705 . In contrast, Ang II-transfected cells, a significant increase in the phosphorylation of tyrosine Stat3 at residue 705, but not serine 727 residue. 3.4.3. Stat3 Bindungsaktivit t. Since tyrosine 705 phosphorylation of Stat3 binding and translocation of nuclear DNA, the effect of angiotensin II on the intracellular Re Bindungsaktivit t is required Stat3DNA was investigated. Mesangial cells were incubated with 5 mM glucose, NG, containing 1 M incubated exogenous or transfected with a