丹酚酸B通过下调细胞凋亡及炎性损伤减轻高糖诱导的RSC96细胞损伤

Salvianolic Acid B Alleviates RSC96 Cell Injury Induced by High Glucose Via Suppressing Cellular Apoptosis and Inflammatory Damage

目的:观察丹酚酸B(salvianolic acid B,Sal B)对高糖(high glucose,HG)诱导的雪旺细胞增殖的影响及其对凋亡和炎性相关基因表达的影响。方法:将体外培养的大鼠雪旺细胞(RSC96细胞)分为对照组(5.6 mmol/L葡萄糖)、HG组(125 mmol/L葡萄糖)、HG+Sal B组(125 mmol/L葡萄糖+1μmol/L Sal B)。作用72 h后,MTT法检测细胞增殖;流式细胞仪检测ROS(reactive oxygen species,ROS)含量;Western blot法检测凋亡及炎性因子相关蛋白Bax(BCL2-Associated X,Bax)、B细胞淋巴瘤-2基因(B-cell lymphoma-2,Bcl-2)、核因子κB P65(nuclear transcription factor kappaB-P65,NF-κB-P65)、白细胞介素1β(Interleukin-1β,IL-1β)的表达。结果:1)MTT结果显示:与对照组比较,HG组OD值明显降低(P<0.05),而HG+Sal B组OD值明显高于HG组(P<0.05),提示HG显著抑制雪旺细胞增殖活性,Sal B可以减轻HG导致的雪旺细胞增殖活性的降低。2)流式结果显示:与对照组相比,HG组ROS含量明显增高(P<0.05),HG+Sal B组ROS含量明显低于HG组(P<0.05),提示HG可显著上调ROS含量,丹酚酸B可下调HG导致的氧化应激过度激活。3)与对照组比较,HG组Bax,P65,IL-1β的蛋白表达水平明显增高,Bcl-2表达水平下调(P<0.05);与HG组比较,HG+Sal B组Bax,P65,IL-1β的蛋白表达降低,Bcl-2表达水平上调(P<0.05),这提示Sal B可抑制细胞凋亡及炎性相关蛋白的表达。结论:Sal B可能通过下调细胞凋亡及炎性损伤减轻HG对RSC96细胞的损伤。

Objective:To observe the influence of Sal B on the proliferation of HG-induced Schwann cells,and its impacts on the expressions of apoptosis and inflammation related genes.Methods:Rat Schwann cells(RSC 96)cultured in vitro were allocated to the control group(5.6mmol/L glucose),HG group(125mmol/L glucose),HG+Sal B group(125mmol/L glucose+1μmol/L Sal B),After acting for 72 hours,MTT method was used to detect cellular proliferation;flow cytometry to measure the contents of ROS,Western blot method to detect the expressions of Bax,Bcl-2,NF-κB-P65 and IL-1β.Results:1)MTT results showed that:compared with the control group,OD value decreased in HG group apparently(P<0.05),while OD value of HG+Sal B group was higher than that of HG group(P<0.05),which suggested that HG could notably inhibit the proliferation activity of Schwann cells,Sal B could relieve the decrease of Schwann cells proliferation activity induced by HG.2)The results of flow cytometry displayed that:compared with the control group,ROS contents increased notably in HG group(P<0.05),ROS contents of HG+Sal B group were lower than these of HG group remarkably(P<0.05),it suggested that HG could significantly up-regulate ROS contents,Sal B could down-regulate over activation of oxidative stress induced by HG.3)compared with the control group,the expressions of Bax,P65 and IL-1βprotein lifted in HG group the expressions of Bcl-2 lowered(P<0.05);compared with HG group,the expressions of Bax,P65 and IL-1βprotein reduced in HG+Sal B group the expressions of Bcl-2 lifted(P<0.05),it showed that Sal B could inhibit the expressions of cellular apoptosis and inflammation-related proteins.Conclusion:Sal B could alleviate the injury of RSC96 cells induced by HG which might be through down regulating cellular apoptosis and inflammatory injury.