METHODS: α-Mangostin (AM) was isolated from C. arborescens and its cell death mechanism was investigated. AM-induced cytotoxicity was observed with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Acridine orange/propidium iodide staining and annexin V were used to detect cells in early phases of apoptosis. High-content screening was used to observe the nuclear condensation, cell permeability, mitochondrial membrane potential, and cytochrome c release. The role of caspases-3/7, -8, and -9, reactive oxygen species, Bcl-2 and Bax expression, and cell cycle arrest were also investigated. To determine the role of the central apoptosis-related proteins, a protein array followed by immunoblot analysis was conducted. Moreover, the involvement of nuclear factor-kappa B (NF-κB) was also analyzed.
RESULTS: Apoptosis was confirmed by the apoptotic cells stained with annexin V and increase in chromatin condensation in nucleus. Treatment of cells with AM promoted cell death-transducing signals that reduced MMP by downregulation of Bcl-2 and upregulation of Bax, triggering cytochrome c release from the mitochondria to the cytosol. The released cytochrome c triggered the activation of caspase-9 followed by the executioner caspase-3/7 and then cleaved the PARP protein. Increase of caspase-8 showed the involvement of extrinsic pathway. AM treatment significantly arrested the cells at the S phase (P<0.05) concomitant with an increase in reactive oxygen species. The protein array and Western blotting demonstrated the expression of HSP70. Moreover, AM significantly blocked the induced translocation of NF-κB from cytoplasm to nucleus.
CONCLUSION: Together, the results demonstrate that the AM isolated from C. arborescens inhibited the proliferation of MDA-MB-231 cells, leading to cell cycle arrest and programmed cell death, which was suggested to occur through both the extrinsic and intrinsic apoptosis pathways with involvement of the NF-κB and HSP70 signaling pathways.
METHODS AND RESULTS: Five groups of rats: normal control, cancer control, TPHE low dose, TPHE high dose and positive control (tamoxifen) were used for the in vivo study. Histopathological examination showed that TPHE significantly suppressed the carcinogenic effect of LA7 tumour cells. The tumour sections from TPHE-treated rats demonstrated significantly reduced expression of Ki67 and PCNA compared to the cancer control group. Using a bioassay-guided approach, the cytotoxic compound of TPHE was identified as a tricyclic sesquiterpene lactone, namely, 8β- hydroxyl- 4β, 15- dihydrozaluzanin C (HDZC). Signs of early and late apoptosis were observed in MCF7 cells treated with HDZC and were attributed to the mitochondrial intrinsic pathway based on the up-regulation of Bax and the down-regulation of Bcl-2. HDZC induced cell cycle arrest in MCF7 cells and increased the expression of p21 and p27 at the mRNA and protein levels.
CONCLUSION: This results of this study substantiate the anticancer effect of TPHE and highlight the involvement of HDZC as one of the contributing compounds that act by initiating mitochondrial-mediated apoptosis.
METHODS: Young healthy volunteers of either sex aged between 19-24 years, participated in the sessions using URNB/ULNB (n = 52) and FURNB/FULNB (n = 28). The nostril dominance was calculated from signals recorded on the PowerLab equipment, representing pressure changes at the end of the nostrils during respiration. The IOP was measured with Tono-Pen. The subjects were divided into 4 groups viz. right nostril dominant (RND), left nostril dominant (LND), transitional right nostril dominant (TRND) and transitional left nostril dominant (TLND) groups. The IOP data 'before and after' URNB/ULNB or FURNB/FULNB were compared by using paired t-test. The baseline data of IOP between the groups were analysed by using independent samples t-test.
RESULTS: The URNB decreased the IOP in the LND and TLND (p < 0.01) and also in the RND (p < 0.05) groups but not significantly in the TRND group. The ULNB decreased the IOP in the RND group (p < 0.01) only. The FURNB significantly reduced the IOP (p < 0.05) only in the LND and RND groups. The FULNB decreased the IOP but not significantly. The baseline IOP did not differ significantly between the LND, RND, TLND and TRND groups.
CONCLUSION: The URNB/FURNB reduced the IOP, while ULNB/FULNB failed to increase the IOP significantly. It is suggested that the lowering of IOP by URNB indicated sympathetic stimulation.