Finally, using rat primary cultured astrocytes, we also demonstrated that TGF scientific assays b1 induces MMP 9 expression in a time dependent manner. The condition media were immunoprecipitated with an anti MMP 9 antibody and analyzed by western Inhibitors,Modulators,Libraries blot. As shown in Figure 8A, TGF b1 induced expression of MMP 9 protein, but not MMP 2 protein, and release into medium, indicating that TGF b1 also induces MMP 9 protein expression and activation in rat primary cultured astrocytes. In addition, pretreatment of rat primary cultured astrocytes with various inhibitors used in RBA 1 cells also significant attenuated TGF b1 induced MMP 9 expression. These data demonstrate that, as in RBA 1 cells, TGF b1 induced MMP 9 expression is also mediated through the same signaling pathways in rat primary culture astrocytes.

Discussion Inhibitors,Modulators,Libraries MMPs contribute to a wide range of biological activities in several CNS diseases, such as stroke, Alzheimers dis ease, and malignant glioma. Among MMPs, MMP 9 expression and activation have been shown to be predo minantly elevated by various brain injuries, sug gesting that MMP 9 may be a critical Inhibitors,Modulators,Libraries molecule in the degradation of ECM and in the pathophysiology of many brain diseases. Another gelatinase, gelatinase A, is constitutively expressed and its expression is usually not inducible in several cell types including brain cells. Moreover, TGF b and related pep tides are simultaneously produced and released follow ing injury to the human CNS. Despite an obviously important role of TGF b in brain trauma and diseases, the processes by which TGF b is implicated in astrocytic functions are not completely understood.

A well established rat astroglial cell line is derived from dissociated cultures of normal neonatal rat brain tissues. According to various analyses in previous studies, the properties of RBA 1 cells are similar to those of normal astrocytes. Thus, we used a culture model of RBA 1 cells Inhibitors,Modulators,Libraries to investigate the mechanisms underlying TGF b1 induced MMP 9 expression and cel lular functional responses. These results suggest that in RBA 1 cells, activation of ROS dependent ERK12 and JNK12 linking to NFB, mediated through a TGF b receptor, is essential for TGF b1 induced MMP 9 gene expression and cell migration. However, previous studies have demonstrated that MMP 2 can be up regulated by some stimuli such as TGF b, but usually participates in development of cancer including growth, invasion, and metastasis.

Abnormal regulation of MAPKs might be implicated in several CNS disorders. Moreover, Inhibitors,Modulators,Libraries TGF b1 has been reported to act as a multifunctional factor through activation of MAPK cascades in different selleckbio cell types. In the present study, we found that ERK12 and JNK12 are required for MMP 9 expression, since RBA 1 cells transfected with dominant negative ERK1, ERK2 or JNK plasmid led to down regulation of MMP 9.