基础医学与临床 ›› 2023, Vol. 43 ›› Issue (4): 615-620.doi: 10.16352/j.issn.1001-6325.2023.04.0615

• 研究论文 • 上一篇    下一篇

miR-140-5p靶向HMGB1/IκB-α轴对糖尿病大鼠视网膜病变的影响

张梦瑶1, 牛姝1, 蔡静1, 赵志刚1*, 周艳2   

  1. 1.石家庄市人民医院 内分泌二科,河北 石家庄 050000;
    2.河北医科大学附属第二医院 检验科,河北 石家庄 050000
  • 收稿日期:2022-04-29 修回日期:2022-11-13 出版日期:2023-04-05 发布日期:2023-04-03
  • 通讯作者: *zhigang_zhao8@126.com
  • 基金资助:
    石家庄市卫生健康委员会资助项目(20190161)

Effect of miR-140-5p targeting HMGB1/IκB-α axis on retinopathy of diabetic rats

ZHANG Mengyao1, NIU Shu1, CAI Jing1, ZHAO Zhigang1*, ZHOU Yan2   

  1. 1. the Second Department of Endocrinology, Shijiazhuang People's Hospital, Shijiazhuang 050000;
    2. Department of Clinical Laboratory, the Second Affiliated Hospital of Hebei Medical University, Shijiazhuang 050000, China
  • Received:2022-04-29 Revised:2022-11-13 Online:2023-04-05 Published:2023-04-03
  • Contact: *zhigang_zhao8@126.com

摘要: 目的 探讨微小RNA(miRNA)miR-140-5p通过靶向调节高迁移率族蛋白1(HMGB1)/核因子κB抑制蛋白α(IκB-α)轴对糖尿病大鼠视网膜病变的影响。方法 将糖尿病大鼠随机分为模型组、miR-140-5p激动剂(miR-140-5p agomir)组、激动剂阴性对照(agomir-NC)组,10只/组,另取10只正常大鼠作为对照组。分离血清及视网膜组织,RT-qPCR检测血清和视网膜组织miR-140-5p、HMGB1 mRNA表达水平;HE染色检测病理学变化;ELISA检测血清单核细胞趋化因子-1(MCP-1)、肿瘤坏死因子-ɑ(TNF-ɑ)、细胞间黏附分子-1(ICAM-1)含量;TUNEL染色检测视网膜细胞凋亡;双荧光素酶报告基因实验检测miR-140-5p、HMGB1的靶向关系;Western blot检测视网膜组织HMGB1、IκB-α、NF-κB p65蛋白表达。结果 HMGB1为miR-140-5p的靶基因。模型组、agomir-NC组MCP-1、TNF-ɑ、ICAM-1含量、HMGB1、NF-κB p65表达、凋亡细胞及组织损伤较对照组增加,miR-140-5p agomir组较模型组降低(P<0.05);模型组、agomir-NC组miR-140-5p、IκB-α表达较对照组降低,miR-140-5p agomir组较模型组增加(P<0.05)。结论 上调miR-140-5p可能通过靶向负调控HMGB1,提高IκB-α表达缓解糖尿病大鼠视网膜病变。

关键词: miR-140-5p, 糖尿病视网膜病变, HMGB1/IκB-α, 细胞凋亡

Abstract: Objective To investigate the effect of miR-140-5p on retinopathy in diabetic rats through targeted regulation of high mobility group protein 1(HMGB1)/nuclear factor κB inhibitory protein-α(IκB-α) axis. Methods Diabetic rats were randomly divided into model group, miR-140-5p agomir group, agomir-NC group, and control group with 10 in each. Serum and retinal tissue were isolated, and the expression of miR-140-5p and HMGB1 mRNA in serum and retinal tissue was detected by RT-qPCR. HE staining was used to evaluate pathological changes. The level of serum monocyte chemoattractant protein-1(MCP-1), tumor necrosis factor-ɑ(TNF-ɑ) and intercellular adhesion molecule-1(ICAM-1) was detected by ELISA. Retinal cell apoptosis was examined by TUNEL staining. Dual luciferase reporter gene assay was used to detect the targeting relationship between miR-140-5p and HMGB1. The protein expressions of HMGB1, IκB-α and NF-κb p65 in retinal tissue were detected by Western blot. Results HMGB1 was the target gene of miR-140-5p. The content of MCP-1, TNF-ɑ, and ICAM-1, the expression of HMGB1 and NF-κB p65 , apoptotic cells counting and tissue damage in the model group and agomir-NC group were found to be all higher than those in the control group. The miR-140-5p agomir group was lower than that in the model group(P<0.05). The expression of miR-140-5p and IκB-α in the model group and agomir-NC group were lower than those in the control group and the miR-140-5p agomir group was higher than that in the model group(P<0.05). Conclusions Up-regulation of miR-140-5p may negatively regulate HMGB1 and improve the expression of IκB-α to alleviate retinopathy of diabetic rats.

Key words: miR-140-5p, diabetic retinopathy, HMGB1/IκB-α, apoptosis

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