Rabbit Anti-BCL2 monoclonal antibody, clone TA21-14

Introduction: With the increasing level of resistance to chemotherapy and radiotherapy and the associated toxicity of these therapies, the uses of plant-based compounds are gaining importance. This study was done to prove the apoptotic potential of Enicostemma axillare in breast cancer cells. Materials and Methods: 3-(4,5-dimethylthiazol-2-yl)-2.5-diphenyl tenazoliinn bromide assay was performed involving the treatment of Michigan Cancer Foundation-7 cells for 24 h with ethanolic extract of E. axillare (EEEA) and doxorubicin to assess the cell viability, lactate dehydrogenase (LDH) was done to assess cytotoxicity, ethidium bromide (EtBr) and acridine orange (AO) assay was done to assess nuclear morphology of apoptotic cells and the expression of proteins associated with apoptosis - Bad, Bcl2, Bax, CytoC, Caspase 3, 8, and 9 were analyzed by western blotting. The data are expressed as mean +/- SEM. Results and Discussion: IC50 value for EEEA is 12.5 mu g/ml, whereas for doxorubicin is 1 mu g/ml, A significant increased level of LDH release is seen in treated groups. EEEA and doxorubicin decrease cell viability. In AO/EtBr staining, the live cells of the EEEA treatment were similar to that of the doxorubicin. Cells stained green represent viable cells, whereas bright red staining represents late apoptotic cells. EEEA down-regulated the expression of Bcl2 (antiapoptotic protein) and up-regulated the expression of Bad and Bax (pioapoptotic proteins) in MCF-7 cell line compared to control cells. In this study, EEEA treatment significantly increased the protein expression of caspase-3, 8, and 9 compared to control. Conclusions: Our finding showed that EEEA induces extrinsic and intrinsic pathway mediated apoptosis. Thus, E. axillare raises new hope for its use in breast cancer therapy.

In this study, we hypothesized that sepsis induction impairs memory retrieval in rats while transplanted mesenchymal stem cells (MSCs) and MSC-conditioned medium (MSC-CM) application are capable of attenuating those complications. MSCs were obtained from adipose tissue of rats and at the second culture passage; MSCs and MSC-CM were collected. Rats were randomly divided into four experimental groups: sham, CLP, MSC, and MSC-CM. Sepsis was induced by cecal ligation and puncture (CLP) model in the CLP, MSC, and MSC-CM groups. The MSC group received 1 x 10(6)MSCs/rat (i.p., 2 h after CLP surgery); the MSC-CM rats received the conditioned medium (CM) from 1 x 10(6)MSCs intraperitoneally 2 h after sepsis induction. Novel object recognition test, sepsis score, and blood pressure measurement were performed 24 h after the treatments. The right hippocampus was taken for western blot analysis. CLP rats showed a significantly higher sepsis score and systolic blood pressure. They also had a significant increase in the phosphorylated form of CAMKII-alpha, cleaved caspase 3 and Bax/Bcl2 ratio, and a reduction in c-fos protein in the hippocampus tissue samples compared with the sham group. MSC transplantation and MSC-CM administration significantly decreased the mean sepsis score and prevented sepsis-induced attenuation of blood pressure compared with the CLP rats. Animals in the MSC and MSC-CM groups showed a better memory retrieval, attenuation in phosphorylated form of CAMKII-alpha, cleaved caspase 3 and Bax/Bcl(2)ratio, and an increase in c-fos protein expression compared with the CLP group. It seems that CAMKII and c-fos are inversely involved in regulating memory processes in hippocampus. Phosphorylated form of CaMKII-alpha overexpression may impair the ability of object recognition. Our findings confirmed that MSC-CM application has more advantages compared with transplanted MSCs and may be offered as a promising therapy for inflammatory diseases such as severe sepsis.