Figure 1
Combination of neuroprotectants and mild hypothermia had better protection effect on primary cultured neurons after OGD/R.
The cultured cortical neurons (DIV 6) were subjected to OGD for 3 hrs. While glucose recovery and reoxygenation initiated, cells were immediately treated with a drug (24 hrs), mild hypothermia (MH, 4.5 hrs at 34°C, then 19.5 hrs at 37°C), or combination of both of them. Cells without OGD/R were as normal control. CVP was measured with the CCK-8 Assay Kit (Dojindo Laboratories, Tokyo, Japan) before OGD (CVpre) and 24 hrs after reoxygenation (CVpost). CVP = CVpost/CVpre × 100%. The bars represent means ± s.d. (n = 6).*, P<0.05 compared with the other groups and **, P<0.05, compared with one or two groups. Among 26 drugs, BDNF, MK-801, GBC, HUK and Ngb, combined with MH presented more efficient protection against OGD/R-induced damage (indicated as b–f). But others had no additional protection with hypothermia, represented by albumin. (a). The CVP of combination of 5% albumin with MH was higher than albumin (t = −4.027, P = 0.002) but not MH alone (t = −0.926, P = 0.376). (b). The CVP of combination of 25 ug/ml BDNF with MH was higher than BDNF (t = −2.564, P = 0.028) or MH alone (t = −4.479, P = 0.001). (c). The CVP of combination of 1 μM GBC with MH was higher than GBC (t = −2.597, P = 0.027) or MH alone (t = −3.210, P = 0.009). (d). The CVP of combination of 0.0015 PNA/ml HUK with MH was higher than HUK (t = −2.484, P = 0.032) or MH alone (t = −3.927, P = 0.003). (e). The CVP of combination of 10 μM MK801 with MH was higher than MK801 (t = −8.064, P<0.001) or MH alone (t = −5.518, P<0.001). (f). The CVP of combination of 50 nM Ngb with MH was higher than Ngb (t = −9.400, P<0.001) or MH alone (t = −5.509, P<0.001). Abbrevation: BDNF: brain-derived neurotrophic factor; GBC: glibenclamide; MK-801: dizocilpine; HUK: human urinary kininogenase; Ngb: neuroglobin; OGD/R: oxygen glucose deprivation and reperfusion; DIV: days in vitro; s.d.: standard deviation; CV: cell viability; CVP: cell viability proportion.
