摘要
目的探讨D-阿洛糖处理对氧糖剥夺再灌注(OGD/R)诱导的HT22细胞损伤及半乳糖凝集素-3(Gal-3)表达的影响。方法正常培养的HT22细胞,传代贴壁后,随机分为对照组、OGD/R组、D-阿洛糖组与抑制剂组。构建OGD/R模型,设立OGD/R组。D-阿洛糖组:构建OGD/R模型,复氧复糖时于培养基中加入10 mmol/L D-阿洛糖,持续到复氧复糖结束。抑制剂组:构建OGD/R模型,复氧复糖时于培养基中同时加入D-阿洛糖和TD139,持续到复氧复糖结束。采用CCK8法检测细胞存活率;Western blot检测Gal-3以及凋亡相关蛋白Bax、Bcl-2、Caspase-3、Cleaved caspase-3的表达;采用流式细胞术检测细胞凋亡。结果与OGD/R组相比,D-阿洛糖组细胞存活率显著升高,细胞凋亡率显著降低,Bax、Cleaved caspase-3、Gal-3、LGALS3BP蛋白表达量显著降低,Bcl-2蛋白表达量显著升高,差异均有统计学意义(P<0.05);而Caspase-3蛋白表达量比较差异无统计学意义(P>0.05)。与D-阿洛糖组相比,抑制剂组的Bcl-2/Bax表达显著增加,Cleaved caspase-3蛋白表达量显著减少,差异均有统计学意义(P<0.05);Caspase-3蛋白表达量比较差异无统计学意义(P>0.05)。结论D-阿洛糖处理对OGD/R诱导的HT22细胞损伤有保护作用以及D-阿洛糖可能通过抑制Gal-3发挥保护作用。
Objective To investigate the effects of D-allose treatment on oxygen glucose deprivation/reperfusion(OGD/R)induced HT22 cell damage and galactin-3(Gal-3)expression.Methods HT22 cells in normal culture were randomly divided into control group,OGD/R group,D-allose group and inhibitor group after passage and adhesion.OGD/R model was constructed and set it as the OGD/R group.D-allose group:OGD/R model was constructed,10 mmol/L D-allose was added into the medium during the reoxygenation and reglucose and continued until the end of the reoxygenation and reglucose.Inhibitor group:OGD/R model was constructed,and D-allose and TD139 were added to the culture medium at the same time during reoxygenation and disaccharide,which lasted until the end of reoxygenation and disaccharide.CCK8 assay was used to detect cell survival rate.The expression of Gal-3 and apoptosis-related proteins Bax,Bcl-2,Caspase-3,Cleaved caspase-3 were detected by Western blot.Apoptosis was detected by flow cytometry.Results Compared with OGD/R group,the survival rate of cells in D-allose group was significantly increased,the apoptosis rate was significantly decreased,the protein expression levels of Bax,Cleaved caspase-3,Gal-3 and LGALS3BP were significantly decreased,and the expression level of Bcl-2 protein was significantly increased,with statistical significance(P<0.05);but there was no significant difference in Caspase-3 protein expression(P>0.05).Compared with the D-allose group,the expression of Bcl-2/Bax in inhibitor group was significantly increased,Cleaved caspase-3 protein expression was significantly decreased,with statistical significance(P<0.05);there was no significant difference in Caspase-3 protein expression(P>0.05).Conclusion D-allose treatment has a protective effect on OGD/R-induced HT22 cell damage,and D-allose may play a protective role by inhibiting Gal-3.
作者
张敏
费晓炜
罗耀文
付奕豪
张磊
高大宽
Zhang Min;Fei Xiaowei;Luo Yaowen;Fu Yihao;Zhang Lei;Gao Dakuan(Department of Neurosurgery,The First Affiliated Hospital of Air Force Medical University,Xi'an 710032,China)
出处
《中华神经创伤外科电子杂志》
2023年第3期135-141,共7页
Chinese Journal Of Neurotraumatic Surgery：Electronic Edition
基金
国家自然科学基金(81971227)。
