[Primary Mechanisms of CD34+CD38---KG1a Leukemia Stem Cells Apoptosis Induced by FA-2-b-ß Extracted from Qinba Selenium- Mushroo].

OBJECTIVE: To investigate the apoptosis of CD34+CD38--KG1a leukemia stem cells induced by Qinba selenium-mushroom extract(FA-2-b-ß), and its related mechanism. METHODS: CD34+CD38---KG1a cells were isolated from KG1a cell line by magnetic activated cell sorting. The proliferation ability of KG1a stem cells treatd by various concentration of FA-2-b-ß(1.2-2.4 mg/ml) in vitro for 24 and 48 hours were tested by cell counting Kit-8(CCK8). Flow cytometry was used to detect the apoptosis rate of KG1a stem cells in each group after treated by FA-2-b-ß in vitro. Expression of BAX,BCL-2,Casepase-3 and Cyclin D1 protein were detected by Western blot. RESULTS: The proportion of CD34+CD38---KG1a stem cells was (95.35±2.63）% after immunomagnetic isolation. The proliferation of KG1a stem cells was inhibited significantly by FA-2-b-ß, which shows a time- and dose-dependent manner (24 h,r=0.943; 48 h,r=0.976). Flow cytometry shows that with the increasing of drug concentration, the apoptosis was also increased, when KG1a stem cells was treated by FA-2-b-ß for 24 h. Western blot indicated that the expression of apoptosis-related protein BAX and Casepase-3 were up-regulated, the expression of BCL-2 and Cyclin D1 were down-regulated. CONCLUSION: FA-2-b-ß can regulate proliferation and apoptosis KG1a stem cells, the involved mechanism may be related with the activation of mitochondrial-mediated apoptotic pathway.

The antidepressant-like effect of human opiorphin via opioid-dependent pathways in mice.

In the present study, we investigated the antidepressive activity of opiorphin with central administration in the forced swim test in mice. Intracerebroventricular (i.c.v.) administration of opiorphin (1-6 µg/mouse) dose-dependently decreased the immobility time, which was reversed by nonselective opioid receptor antagonist naloxone, δ-selective naltrindole and µ-selective ß-FNA. The data suggested that central administration of opiorphin produced an antidepressant-like effect by activating both µ and δ opioid receptors indirectly. In order to eliminate the possibility of a false-positive result in the forced swim test, locomotor activity was checked in both non-habituated and habituated mice. Opiorphin had no influence on non-habituated mice, though had weak effect on habituated mice. In addition, mice treated with opiorphin did not display any convulsive behaviors.

Central administration of neuronostatin induces antinociception in mice.

Neuronostatin, a recently discovered endogenous bioactive peptide, was encoded by pro-mRNA of somatostatin that contributes to modulation of nociception. However, nociceptive effect of neuronostatin is still not fully known. The aim of this study was to evaluate effect of neuronostatin on nociception and elucidate its possible mechanism of action. Intracerebroventricular (i.c.v.) administration of neuronostatin (0.3, 3, 6, 12nmol/mouse) produced a dose- and time-related antinociceptive effect in the tail immersion assay in mice, an acute pain model. The antinociceptive effect of neuronostatin was significantly antagonized by naloxone, and was strongly inhibited by co-injection with ß-funaltrexamine or nor-binaltorphimine, but not by naltrindole. Also, melanocortin 3/4 receptor antagonist, SHU9119, completely blocked the effect of neuronostatin. These data indicated the involvement of both µ- and κ-opioid receptors and central melanocortin system in the analgesic response induced by neuronostatin. In addition, neuronostatin (6nmol, i.c.v.) increased c-Fos protein expression in the periaqueductal gray (PAG) and the nucleus raphe magnus (NRM) that have a pivotal role in regulating descending pain pathways. Taken together, this study is the first to reveal that neuronostatin produces antinociceptive effect via opioid and central melanocortin systems, which is associated with an increase in neuronal activity the PAG and NRM.