ISSN 1004-6879

CN 13-1154/R

 

承德医学院学报 ›› 2022, Vol. 39 ›› Issue (3): 186-192.

• 基础医学 • 上一篇    下一篇

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

胡万祥, 李金尧, 李雨欣, 韩思雨, 程露阳*, 陈志宏*   

  1. 承德医学院基础医学院,河北承德 067000
  • 收稿日期:2021-10-25 出版日期:2022-06-10 发布日期:2022-08-02
  • 通讯作者: *
  • 基金资助:
    承德医学院国家自然科学基金项目培育基金(201912)

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

HU Wan-xiang, LI Jin-yao, LI Yu-xin, HAN Si-yu, CHENG Lu-yang*, CHEN Zhi-hong*   

  1. School of Basic Medicine, Chengde Medical University, Chengde, Hebei, 067000, China
  • Received:2021-10-25 Online:2022-06-10 Published:2022-08-02

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。

关键词: 丝胶, 2型糖尿病, INS-1细胞, 氧化应激, 自噬

Abstract: 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). Conclusions ericin 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.

Key words: sericin, type 2 dibaetes mellitus, INS-1 cells, autophagy, oxidative stress

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