西格列汀对高糖环境下胎鼠海马神经元的影响  

Effects of sitagliptin on hippocampal neuronal apoptosis in high glucose environment

作　　者：熊丽娇[1,2] 陈丽莉 蔡霞[3] 邹玲婷 万绍媛 王萍 许荣[1] 曾治平[1,2] Xiong Lijiao;Chen Lili;Cai Xia;Zou Lingting;Wan Shaoyuan;Wang Ping;Xu Rong;Zeng Zhiping(The First Affiliated Hospital of Gannan Medical University,Ganzhou 341000,China;Jiangxi Branch Center of National Geriatric Disease Clinical Medical Research Center,Ganzhou 341000,China;Gannan Healthcare Vocational College,Ganzhou 341000,China)

机构地区：[1]赣南医学院第一附属医院,赣州341000 [2]国家老年疾病临床医学研究中心江西分中心,赣州341000 [3]赣南卫生健康职业学院,赣州341000

出　　处：《国际内分泌代谢杂志》2021年第6期617-622,共6页International Journal of Endocrinology and Metabolism

基　　金：江西省教育厅科技项目(GJJ201545);赣南医学院科研课题(YB201808);赣州市指导性科技计划(GZ2018ZSF010);赣南医学院第一附属医院院级科技计划(YJZD202011)。

摘　　要：目的观察西格列汀对高糖环境下胎鼠海马神经元的影响。方法原代胎鼠海马神经元细胞培养7 d,分为对照组(Con组)、高糖模型组(Mod组)、西格列汀组(Sita组)和甘露醇组(MT组)。Con组不处理,Sita组及Mod组使用高糖环境(100 mmol/L),Sita组加入100μmol/L西格列汀,MT组加入4.5%甘露醇,处理24 h。应用噻唑蓝(MTT)法测细胞活力,原位末端转移酶标记技术(TUNEL)染色及膜连蛋白V/碘化丙啶(AnnexinV-FITC/PI)流式细胞术测细胞凋亡,蛋白质免疫印迹技术测定B细胞淋巴瘤/白血病-2基因(Bcl-2)、Bcl-2相关X蛋白基因蛋白(Bax)、含半胱氨酸的天冬氨酸蛋白水解酶-3(Caspase-3)的表达,测定丙二醛(MDA)含量及超氧化物歧化酶(SOD)活性。结果MT组存活率、凋亡率、MDA、SOD及各蛋白表达与对照组比差异无统计学意义(P>0.05)。Mod组较Con组海马神经元细胞存活率下降(P<0.05),凋亡率升高(P<0.05),Bax、Caspase-3表达和MDA含量增加(P均<0.05),Bcl-2表达和SOD活力降低(P均<0.05)。Sita组较Mod组存活率上升(P<0.05),凋亡率下降(P<0.05),Caspase-3、Bax表达和MDA含量降低(P<0.05),Bcl-2表达和SOD活力增加(P均<0.05)。结论在细胞水平西格列汀可抑制高糖环境下海马神经元细胞凋亡和氧化应激,具有神经保护作用。Objective To observe the effects of sitagliptin on hippocampal neurons in fetal rats under a high glucose environment.Methods Primary fetal hippocampal neurons were cultured for 7 days and divided into control group(Con group),high glucose model group(Mod group),sitagliptin group(Sita group)and mannitol group(MT group).The Con group was not treated.The Sita group and the Mod group were treated with high glucose(100 mmol/L).The Sita group was treated with 100μmol/L sitagliptin.MT group was treated with 4.5%mannitol.All of them were treated for 24 h.Cell viability was measured by methyl thiazolyl tetrazolium(MTT)method,and apoptosis rate was measured by Terminal-deoxynucleoitidyl Transferase Mediated Nick End Labeling(TUNEL)staining and AnnexinV-FITC/PI flow cytometry.B-cell lymphocyte/leukemia-2(Bcl-2),Bcl-2 associated X protein gene protein(Bax),and Caspase-3 protein expression were measured by Western-blot method.Malondialdehyde(MDA)content and superoxide dismutase(SOD)activity were measured.Results There were no significant differences in apoptosis rate,MDA,SOD and protein expression between MT group and Con group(all P>0.05).Compared with Con group,the cell viability of hippocampal neurons in Mod group was decreased(P<0.05)and the apoptosis rate was increased(P<0.05).Caspase-3,Bax expression and MDA content in Mod group were increased compared with Con group(all P<0.05),and the expressions of Bcl-2 and SOD activity were decreased(all P<0.05).The survival rate of Sita group was higher than that of Mod group(all P<0.05),and the apoptosis rate was lower(P<0.05).Caspase-3,Bax expression and MDA content in Sita group were significantly lower than those in Mod group(P<0.05).In contrast,Bcl-2 expression and SOD activity expressions were substantially higher than those in Mod group(all P<0.05).Conclusion Sitagliptin could play a neuroprotective role by inhibiting apoptosis and oxidative stress in hippocampal neurons under high glucose environment at the cellular level.

关 键 词：西格列汀 糖尿病 海马神经元 细胞凋亡 氧化应激 

分 类 号：R587.2[医药卫生—内分泌]