S14G-humanin对氧糖剥夺/复氧神经细胞损伤的保护作用

目的：评估S14G-humanin(HNG)在SH-SY5Y神经母细胞瘤细胞体外氧糖剥夺/复氧(oxygen glucose deprivation/reoxygenation,OGD/R)模型中潜在的神经保护作用的相关分子机制。方法：将SH-SY5Y细胞分为对照组、OGD/R模型组、HNG干预组、HNG联合FLLL32抑制剂组。三气培养箱建立SH-SY5Y细胞OGD/R损伤模型；抑制剂组给予不同剂量HNG、FLLL32(JAK2/STAT3通路抑制剂)。CCK8细胞检测试剂盒检测细胞活性，流式细胞仪检测细胞凋亡率，Western Blot分析JAK2、p-STAT3(Y705)、p-STAT3(S727)的表达。结果：OGD/R导致SH-SY5Y细胞存活率显著降低(P<0.01)，凋亡率显著增加(P<0.01)。给予0.1、1、5、10μg·L^-1 HNG干预的SH-SY5Y细胞与未行HNG干预的OGD/R细胞相比，存活率更高(P<0.01)，凋亡明显减少(P<0.01),1μg·L^-1HNG干预组效果最好。与对照组...

Neuroprotective Effect of S14G-Humanin on SH-SY5Y Cells induced by Oxygen Glucose Deprivation/Reoxygenation

Objective：To establish an in vitro oxygen glucose deprivation/reoxygenation（OGD/R） model using SH-SY5Y neuroblastoma cells to mimic the in vitro ischemia/reperfusioninjury in stroke. To evaluate the potential neuroprotective effect of S14G-humanin （HNG） fromischemia/reperfusion in vitro，and further to identify the molecular mechanism underlying theprotection，so as to provide a theoretical basis for the treatment of cerebral ischemia/reperfusioninjury with HNG. Methods：SH-SY5Y cells were divided into control group，oxygen-glucose deprivation reperfusion OGD/R model group and inhibitor group. The present study establishedan in vitro OGD/R model using SH-SY5Y neuroblastoma cells by three gas incubator. HNG andFLLL32，an inhibitor of JAK2/STAT3 signal，were added to the cells in indicated experiments.CCK8 kit was used to detect the cell viability，and flow cytometry was used to detect the cellapoptosis rate. The expression of JAK2，p-STAT3（Y705） and p-STAT3（S727） were analyzedby Western Blot. Results：The results showed that OGD/R resulted in a significant decrease inthe survival rate of SH-SY5Y cells （P<0.01） and a significant increase in the apoptosis rate ofSH-SY5Y cells （P<0.01）. SH-SY5Y cells incubated 0.1，1，5 or 10 μg·L^-1 HNG showed highercell survival （P<0.01） and significantly decreased apoptosis （P<0.01） compared with the cellswithout HNG treatment in OGD/R processes，with the most evident effects at 1 μg·L^-1 HNG.OGD/R lowered protein levels of JAK2 （P<0.01） and p-STAT3 at Y705 site （P<0.01），comparedto control. Adding HNG to the cells increased protein levels of JAK2 （P<0.01） and p-STAT3（Y705）（ P<0.01） in OGD/R conditions. Moreover，protein levels of JAK2（ P<0.01），p-STAT3（Y705）（ P<0.01） and p-STAT3（ S727）（ P<0.05） had notable increase at values of 1 and 5 μg·L^-1 HNG in OGD/R conditions. Co-treatment with HNG and FLLL32 was unable to restore the cellviability and abolishing the inhibition of apoptosis by HNG（P >0.05）. HNG inhibited OGD/R-induced reduction of JAK2，p-STAT3（Y705） and p-STAT3（S727） protein expression，whereas the inhibitory effect was not observed with co-treatment with HNG and FLLL32. Conclusion：These data collectively indicate that HNG has neuroprotective effects against OGD/R by activating JAK2/STAT3 signaling and suggest that HNG may be a promising drug for thetreatment of stroke.