丝胶通过影响自噬和氧化应激发挥保护STZ致胰岛素瘤细胞损伤的作用  

Sericin Protects INS-1 Cells from STZ Injury by Affecting Autophagy and Oxidative Stress

作　　者：胡万祥 李金尧 李雨欣 韩思雨 程露阳 陈志宏 HU Wan-xiang;LI Jin-yao;LI Yu-xin;HAN Si-yu;CHENG Lu-yang;CHEN Zhi-hong(School of Basic Medicine,Chengde Medical University,Chengde,Hebei,067000,China)

机构地区：[1]承德医学院基础医学院,河北承德067000

出　　处：《承德医学院学报》2022年第3期186-192,共7页Journal of Chengde Medical University

基　　金：承德医学院国家自然科学基金项目培育基金(201912)。

摘　　要：目的探讨丝胶是否通过影响自噬和氧化应激发挥保护链脲佐菌素(STZ)致胰岛素瘤细胞(INS-1细胞)损伤的作用。方法INS-1细胞随机分为A、B、C、D、E五组。A组(正常组)细胞常规条件培养24h,不做其他处置;B组(STZ致损伤组)细胞中加入10mmol/L的STZ培养24h;C组、D组、E组细胞中分别加入10mmol/L的STZ和不同剂量的丝胶(150μg/mL、300μg/mL、600μg/mL)培养24h。分别采用免疫蛋白印迹法和实时荧光定量PCR法检测细胞自噬相关基因蛋白和mRNA表达,DCFH-DA活性氧(ROS)荧光探针检测细胞ROS的含量。结果C组、D组、E组LC3B-II/LC3B-I、SIRT1、ATG5、BECLIN1、PINK1蛋白和mRNA的表达均明显高于B组(P<0.05);3组细胞各指标蛋白和mRNA的表达水平依次为E组>D组>C组,差异有统计学意义(P<0.05)。C组、D组、E组INS-1细胞ROS含量明显低于B组(P<0.05);3组细胞ROS含量依次为E组<D组<C组,差异有统计学意义(P<0.05)。结论丝胶具有保护STZ致INS-1细胞损伤的作用,其保护机制与丝胶能减轻氧化应激和增强自噬作用有关。Objective To investigate whether sericin can protect streptozotocin(STZ)induced INS-1 cells injury by affecting autophagy and oxidative stress.Methods INS-1 cells were randomly divided into group A,B,C,D and E.In group A(normal group),INS-1 cells were cultured under conventional conditions for 24h without other treatments.In group B(STZ induced injury group),INS-1 cells were cultured with 10 mmol/L STZ for 24h.In group C,D and E,INS-1 cells were respectivly cultured with 10mmol/L STZ and different dose sericin(150μg/mL,300μg/mL,600μg/mL)for 24h.Western blot and real-time fluorescence quantitative PCR were used to detect the expressions of autophagy related gene protein and mRNA,and the reactive oxygen species(ROS)content in INS-1 cellswas detected by DCFH-DA ROS fluorescence probe.Results The expressions of LC3B-II/I,SIRT1,ATG5,BECLIN1,PINK1 proteins and mRNA in group C,D and E were all significantly higher than those in group B(P<0.05).The expression levels of each index in the three groups were group E>group D>group C,and the differences were statistically significant(P<0.05).The ROS content of INS-1 cells in group C,D and E were all obviously lower than that in group B(P<0.05).The ROS content of the three groups was in the order of group E<group D<group C,and the difference was statistically significant(P<0.05).Conclusion Sericin can protect STZ induced INS-1 cells injury,and the protective mechanism is related to sericin’s ability to reduce oxidative stress and enhance autophagy.

关 键 词：丝胶 2型糖尿病 INS-1细胞 氧化应激 自噬 

分 类 号：R587.1[医药卫生—内分泌]