河北医科大学学报 ›› 2024, Vol. 45 ›› Issue (12): 1374-1379.doi: 10.3969/j.issn.1007-3205.2024.12.003

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HYOU1在糖尿病施万细胞凋亡中的作用及对JAK-STAT通路的影响研究

  

  1. 1.河北医科大学大型科研仪器设备共享服务平台,河北 石家庄 050017; 2.河北医科大学基础医学院神经生物研究室,
    河北 石家庄 050017; 3. 河北医科大学基础医学院病理学教研室,河北 石家庄 050017;
    4.河北医科大学教学实验中心,河北 石家庄 050017

  • 出版日期:2024-12-25 发布日期:2025-01-03
  • 作者简介:周晨明(1984-),男,河北保定人,河北医科大学实验师,医学硕士,从事细胞凋亡研究。
  • 基金资助:
    河北省自然科学基金资助项目(H2021206086);河北省医学科学研究课题计划(20220974)

Effect of HYOU1 on apoptosis of diabetic Schwann cells and its effect on JAK-STAT pathway

  1. 1.Core Facilities and Centers, Hebei Medical University, Shijiazhuang 050017, China; 2.Department of 
    Neurobiology, Basic Medical College, Hebei Medical University, Shijiazhuang 050017, China; 
    3.Department of Pathology, Basic Medical College, Hebei Medical University, Shijiazhuang 
    050017, China; 4.Department of Teaching  and Experiment Center, Hebei Medical 
    University, Shijiazhuang 050017, China

  • Online:2024-12-25 Published:2025-01-03

摘要: 目的 探讨内质网分子伴侣复合物缺氧上调基因1(hypoxia up-regulated 1,HYOU1)对糖尿病施万细胞凋亡的影响及其机制。
方法 将对数生长期的施万细胞RSC96使用高糖培养基构建糖尿病周围神经病变细胞模型,并转染过表达HYOU1的质粒。利用RT-qPCR、Western blot检测HYOU1在RSC96细胞株的表达情况;利用CCK8、荧光显微镜、透射电镜和流式细胞术等技术检测HYOU1对RSC96细胞凋亡的影响;利用Western blot检测HYOU1对凋亡相关蛋白的影响,同时检测JAK-STAT信号通路相关蛋白,判断其对该通路的影响。
结果 RT-qPCR和Western blot实验结果显示,过表达HYOU1的RSC96细胞株成功构建。CCK8、荧光显微镜、透射电镜和流式细胞术结果表明,过表达HYOU1后,细胞活力显著升高,细胞凋亡数量显著减少(P<0.05)。Western blot实验结果表明,过表达HYOU1后,抗凋亡因子显著增加,促凋亡因子显著减弱,同时抑制了JAK-STAT通路的激活(P<0.05)。
结论 HYOU1可通过抑制JAK-STAT通路从而影响高糖诱导的RSC96细胞的抑凋亡作用。


关键词: 糖尿病, 施万细胞, JAK-STAT信号通路

Abstract: Objective To explore the effects of the endoplasmic reticulum molecular chaperone complex hypoxia up-regulated gene 1 (HYOU1) on apoptosis in diabetic Schwann cells and its mechanisms. 
Methods A diabetic peripheral neuropathy cell model was constructed from logarithmic growth phase Schwann cells RSC96 using high sugar medium and transfected with a plasmid overexpressing HYOU1. The expression of HYOU1 in RSC96 cell line was detected by RT-qPCR and Western blot. The effect of HYOU1 on the apoptosis of RSC96 cells was detected by CCK8, fluorescence microscopy, transmission electron microscopy and flow cytometry. The effects of HYOU1 on apoptosis-related proteins and JAK-STAT signaling pathway were detected by Western blot. 
Results The results of RT-qPCR and Western blot assay indicated that the RSC96 cell line overexpressing HYOU1 had been successfully constructed. CCK8, fluorescence microscopy, transmission electron microscopy and flow cytometry results showed that overexpression of HYOU1 resulted in a significant increase in cell viability and a significant decrease in the number of apoptotic cells (P<0.05). Western blot results indicated that after overexpression of HYOU1, the anti-apoptotic factors increased significantly, the pro-apoptotic factors decreased significantly, and the activation of JAK-STAT pathway was inhibited (P<0.05). 
Conclusion HYOU1 may inhibit the apoptosis of RSC96 cells induced by high glucose by inhibiting JAK-STAT pathway. 


Key words: diabetes mellitus, Schwann cells, JAK-STAT signaling pathway