荔枝核总皂苷对脂多糖诱导的神经细胞损伤的影响研究

Effect of Lychee seed saponins on nerve cell damage induced by LPS

目的:探讨荔枝核总皂苷(LSS)对脂多糖(LPS)诱导的PC12细胞活力、细胞凋亡率及免疫因子的影响及机制。方法:3. 75、7. 5、15、30、60 mg/L的LSS处理PC12细胞24 h,MTT检测细胞存活率;随机将PC12细胞分为对照组(细胞不做任何处理)、LPS处理组(1 mg/ml的LPS处理PC12细胞24 h)、LSS处理组(3. 75 mg/L的LSS处理细胞24 h后,再用1 mg/ml的LPS处理PC12细胞24 h)。MTT及流式细胞术分别检测细胞存活率及凋亡率; H2DCFHDA荧光探针检测ROS含量;Western blot检测PI3K/AKT信号通路PI3K、p-AKT和下游cycyin D1、Bax及免疫抑制因子HIF-1α和VEGF的蛋白表达。结果:3. 75 mg/L和7. 5 mg/L的LSS对PC12细胞存活率的影响与0 mg/L LSS比较差异无统计学意义(P>0. 05),而15 mg/L、30 mg/L和60 mg/L的LSS均能显著降低PC12细胞存活率,与0 mg/L LSS比较差异具有统计学意义(P <0. 05)。选择3. 75 mg/L的LSS为研究对象;与对照组比较,LPS组细胞存活率显著降低,细胞凋亡率显著升高,ROS含量显著升高,PI3K、p-AKT和cycyin D1的蛋白表达显著降低,Bax、HIF-1α和VEGF的蛋白表达显著升高(P<0. 05)。与LPS组比较,LSS组细胞存活率显著升高,细胞凋亡率显著降低,ROS含量显著降低,PI3K、p-AKT和cycyin D1的蛋白表达显著升高,Bax、HIF-1α和VEGF的蛋白表达显著降低(P<0. 05)。结论:LSS可提高PC12细胞存活率,降低细胞凋亡,提高免疫,其机制与激活PI3K/AKT信号、提高细胞ROS水平及抑制HIF-1α和VEGF的表达有关。

Objective: To investigate the effect of Lychee seed saponins( LSS) on the cell activity,cell apoptosis rate and immune factors of PC12 cells induced by lipopolysaccharide( LPS). Methods: 3. 75,7. 5,15,30 and 60 mg/L LSS treated with PC12 cells for 24 h,and MTT was used to detect cell survival. PC12 cells were randomly divided into control group( cells without any treatment),LPS treatment group( 1 mg/ml LPS treated with PC12 cell for 24 h),LSS treatment group( After 3. 75 mg/L LSS treated cell for 24 h,the PC12 cell was treated with 1 mg/ml LPS for 24 h). MTT and flow cytometry were used to detect cell survival rate and apoptosis rate respectively. H2 DCFHDA fluorescence probe was used to detect ROS content;Western blot was used to detect protein expression of PI3 K,p-AKT,cycyin D1,Bax,HIF-1α and Bax. Results: The effect of 3. 75 mg/L and 7. 5 mg/L LSS on the survival rate of PC12 cells was not statistically different compared with 0 mg/L LSS( P>0. 05),while the 15 mg/L,30 mg/L and 60 mg/L LSS could significantly reduce the survival rate of the PC12 cells,and there was statistically significant difference compared with 0 mg/L( P >0. 05). 3. 75 mg/L’s LSS was chosen as the research object. Compared with the control group,cell survival rate decreased significantly in LPS group,the apoptosis rate was significantly increased,ROS content increased significantly,the expression of PI3 K,p-AKT and cycyin D1 protein was significantly decreased,the expression of Bax,HIF-1α and VEGF protein increased significantly( P < 0. 05).Compared with LPS group,the survival rate was significantly increased in LSS group,cell apoptosis rate was significantly decreased,ROS content was significantly reduced,PI3 K,p-AKT and cycyin D1 protein expression increased significantly,Bax,HIF-1α and VEGF protein expression decreased significantly( P < 0. 05). Conclusion: LSS can increase the survival rate of PC12 cells,decrease cell apoptosis and enhance immunity. Its mechanism is related to activation of PI3 K/AKT signal,increase of ROS level and inhibition of HIF-1α and VEGF expression.