[Hypoxic preconditioning increases cPKCgamma membrane translocation in murine brain].

Cerebral hypoxic preconditioning (CHP), which was induced by repetitive sub-lethal hypoxic insult, is an endogenous protection of neuron against subsequent severe hypoxic injury. Although a number of possible induction pathways have been investigated, such as neuroactive cytokines, activation of glutamate receptors, the ATP-sensitive potassium channel, nitric oxide and oxidative stress, the exact mechanism underlying CHP-induced protection remains unclear. It is interesting that all the above-mentioned mechanisms are involved in the activation of protein kinases C (PKC). Recently we reported that the level of PKCs membrane translocation was significantly increased in the brain of hypoxic preconditioned mice. In order to explore the role of conventional protein kinases C (cPKC) in the development of cerebral hypoxic preconditioning, biochemical techniques of SDS-PAGE and Western bolt were applied to observe the effects of repetitive hypoxic exposure (H1-H4) on the level of cPKCalpha and gamma membrane translocation in the cortex and hippocampus of mice. Experiments were carried out in accordance with the National Institutes of Health guide for the care and use of laboratory animals. The hypoxic preconditioned mice model was adapted with minor modification from our previous report. In brief, healthy adult BALB/C mice weighing 18-20 g of either sex were randomly divided into 5 groups: control group (H0), hypoxic control group (H1, hypoxic exposure once ), hypoxic preconditioned group (H2-H4, repetitive hypoxic exposure for 2-4 times respectively). The first sign of gasping breath was taken as the end of each hypoxic exposure, and then the mice were kept in normal control condition for a 30-min interval to recover before the following hypoxic insult. We found that the level of cPKCgamma membrane translocation was increased significantly (*P<0.05, n=6) with the increase of the hypoxic exposure times in both hippocampus (H0: 100% vs H1 approximately H4: 119.2%+/-7.0% *, 139.3% +/-7.4%*, 134.2% +/-8.95%*, 184.0% +/-10.8%*) and cortex (H0: 100% vs H1-H4: 129.7% +/-13.8%, 143.3% +/-13.9%*, 204.0% +/-12.1%*, 229.5% +/-14.6%*) of mice. But there were no significant changes in cPKCalpha membrane translocation in cortex and hippocampi of hypoxic preconditioned mice. These results suggest that cPKCgamma plays an important role in the development of cerebral hypoxic preconditioning. The changes in some other forms of novel and atypical PKCs are still under investigation.

[Hypoxic preconditioning increase nPKCepsilon membrane translocation in the brain of mice].

AIM: To explore the role of novel protein kinases C (nPKCs) in the development of cerebral hypoxic preconditioning. METHODS: By using the mice model of hypoxic preconditioning, which was established before in our lab, the biochemistry techniques of SDS-PAGE and Western blot were applied to observe the effects of repetitive hypoxic exposure (H0-H4) on nPKCs (nPKCepsilon, delta, eta, mu and theta) membrane translocation in hippocampus and cortex. RESULTS: nPKCepsilon membrane translocation was increased in response to the hypoxic exposure times in the hippocampus (H0: 41.6% +/- 1.4% vs. H1-H4: 46.9% +/- 4.5%, 52.7% +/- 3.9%, 58.8% +/- 2.7% and 61.3% +/- 3.7%) and cortex (H0: 38.4% +/- 4.5% vs. 42.4% +/- 5.0%, 48.7% +/- 6.5%, 55.3% +/- 8.9% and 61.2% +/- 10.2%) of mice, and there were statistic significances among H2, H3 and H4 in hippocampus, and H3 and H4 in cortex respectively (P < 0.01). But for nPKCdelta, eta, mu and theta membrane translocation, there were no any significant changes in hippocampus and cortex of hypoxic preconditioned mice. CONCLUSION: nPKCepsilon may play an important role in the development of cerebral hypoxic preconditioning, but it need more evidence to prove.