Since the higher BMDMC pulmonary engraftment observed with intratracheal instillation compared to intravenous injection did not potentiate the beneficial effects of BMDMC therapy, these beneficial changes may be attributed to the ability of BMDMCs to modulate IL-4, IL-13, TGF-β and VEGF levels in lung tissue from a distant site. In the present study, we used a model of allergic inflammation previously described by our group in BALB/c

mice (Xisto et al., 2005, Burburan et al., 2007 and Antunes et al., 2009). Nevertheless, C57BL/6 mice were used, because they serve as a background selleck compound strain for GFP mice (Abreu et al., 2011a) and exhibit inflammatory (eosinophilia and Th2 pro-inflammatory cytokine increase) and ultrastructural changes in the airway and lung parenchyma which closely mirror human disease compared to other strains, even in the absence of alum adjuvant (Yu et al., 2006, Antunes et al., 2009 and Allen et al., 2012). A recent study demonstrated that NLRP3 inflammasome activation is essential in alum-free models of allergic asthma as it leads to IL-1 production,

a critical factor for the induction of Th2 inflammatory allergic response (Besnard et al., 2011). BMN 673 manufacturer Even though the use of alum adjuvant during the immunization phase of the OVA model has been demonstrated to enhance the cardinal

features of allergic airway disease, this practice has been called into question, since it is an artificial method of asthma induction with major differences in relation to the pathogenesis of allergic disease in humans. Several recent studies have investigated the intravenous administration of mesenchymal stem cells in experimental models of asthma, focusing on the beneficial effects of these cells on lung remodelling and inflammation (Bonfield et al., 2010, Firinci et al., 2011 and Goodwin et al., 2011). However, MSC pose a from series of disadvantages, such as culture conditions detrimental to cell transplantation and risk of contamination and immunologic reactions. In light of these limitations, our group evaluated the effects of intravenous BMDMC administration in a model of allergic asthma (Abreu et al., 2011a). BMDMCs can be administered easily and safely on the day of harvesting. They also express several genes involved in inflammatory response and chemotaxis (Ohnishi et al., 2007), and are less costly than MSCs. Additionally, further studies should investigate whether the nature of BMDMCs as an heterogeneous mix of progenitor and immune cells could induce beneficial effects, with each cellular type playing a specific role.

Fig. 3 and Table 1 depict that the IC50 values markedly decreased with the addition

of SG to epirubicin and paclitaxel. The IC50 value of epirubicin in the HeLa cells was 1.05 μg/mL, which decreased to 0.15 μg/mL with the addition of 80 μg/mL SG. This result indicates that a subtoxic concentration of SG significantly increases the cytotoxic efficacy of epirubicin. SG exhibited similar check details potentiating activities on paclitaxel in all three cancer cell lines. To examine whether the role of SG in the cytotoxic effect of epirubicin and paclitaxel was caused by the enhanced apoptosis, we assessed the resulting apoptosis in the HeLa cells after separate treatments with epirubicin and paclitaxel alone and after the treatment with the combination of SG and the two drugs. The stage of apoptosis was determined through annexin-V analysis. As shown in Fig. 4A and C, the percentage of apoptotic cells was considerably higher in the cotreated cells than in the epirubicin- and paclitaxel-treated cells. To determine the activation Tofacitinib of caspase in the cells, we detected the PARP cleavage through immunoblotting analysis.

Fig. 4B and D show that PARP was cleaved to yield an 85-kD fragment in the drug-treated cells and that the amount of the cleaved 85-kD fragment was more significant in the co-treated cells than in the epirubicin- and paclitaxel-treated 4-Aminobutyrate aminotransferase cells. On the basis of these results, we suggest that SG enhances the anticancer activities of epirubicin and paclitaxel through caspase-associated apoptosis. To elucidate the initiation event of apoptosis, we inspected the activation kinetics of the two initiator caspases, namely, caspase-8 and -9, and the effector caspases, caspase-3/-7. As shown in Fig. 5,

the activities of caspase-9 and -3/-7 greatly increased in the cotreated cells than in the epirubicin- and paclitaxel-treated cells. By contrast, the activity of caspase-8 did not show any change in all cells. We then determined the cleavage of caspase-9 and -8. Specifically, we examined the proteolytic activation of these caspases through immunoblotting analysis. Apparent cleavage was observed in caspase-9 but not in caspase-8. The amounts of the active form of the cleaved caspase-9 were higher in the cotreated cells than in the epirubicin- and paclitaxel-treated cells. The data suggest that epirubicin and paclitaxel-induced apoptosis might be potentiated by SG via the intrinsic apoptosis pathway in HeLa cells. The release of mitochondrial cytochrome c is the crucial event in caspase-9 activation [40]. The family members of the Bcl-2 family, namely, Bax and Bak, serve as an essential gateway for the release of cytochrome c [5] and [41]. Fig.