[1]焦军杰,姚文艳,张前辉,等.转甲状腺素蛋白介导STAT4/miR-223-3p/FBXW7通路对高糖诱导的人视网膜内皮细胞新生血管生成的影响[J].眼科新进展,2023,43(2):099-104.[doi:10.13389/j.cnki.rao.2023.0020]
　JIAO Junjie,YAO Wenyan,ZHANG Qianhui,et al.Effect of the transthyretin on high glucose-induced angiogenesis of human retinal endothelial cells via mediating the signal transducer and activator of transcription 4/miR-223-3p/F-box and WD repeat domain containing 7 pathway[J].Recent Advances in Ophthalmology,2023,43(2):099-104.[doi:10.13389/j.cnki.rao.2023.0020]
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转甲状腺素蛋白介导STAT4/miR-223-3p/FBXW7通路对高糖诱导的人视网膜内皮细胞新生血管生成的影响/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
43卷
期数:
2023年2期
页码:
099-104
栏目:
实验研究
出版日期:
2023-02-05

文章信息/Info

Title:
Effect of the transthyretin on high glucose-induced angiogenesis of human retinal endothelial cells via mediating the signal transducer and activator of transcription 4/miR-223-3p/F-box and WD repeat domain containing 7 pathway
作者:
焦军杰; 姚文艳; 张前辉; 李秀娇; 常昆; 马萧萧; 李晓鹏
453003　河南省新乡市，新乡医学院第三附属医院
Author(s):
JIAO Junjie; YAO Wenyan; ZHANG Qianhui; LI Xiujiao; CHANG Kun; MA Xiaoxiao; LI Xiaopeng
Department of Ophthalmology,the Third Affiliated Hospital of Xinxiang Medical University,Xinxiang 453003,Henan Province,China
关键词:
转甲状腺素蛋白; 人视网膜内皮细胞; 糖尿病视网膜病变; 信号转导和转录激活因子4; miR-223-3p; F-box WD 重复蛋白 7
Keywords:
transthyretin;  human retinal endothelial cells;  diabetic retinopathy;  signal transducer and activator of transcription 4;  miR-223-3p;  F-box and WD repeat domain containing 7
分类号:
R774.1
DOI:
10.13389/j.cnki.rao.2023.0020
文献标志码:
A
摘要:
目的　分析转甲状腺素蛋白（TTR）介导信号转导和转录激活因子4(STAT4）/miR-223-3p/ F-box WD重复蛋白7（FBXW7）通路对高糖（HG）诱导的人视网膜内皮细胞（hRECs）新生血管生成的影响。方法　将hRECs分为对照组、HG组、HG+TTR组、HG+NC mimic组、HG+miR-223-3p mimic组和HG+TTR+miR-223-3p mimic组。生物信息学分析STAT4与miR-223-3p的靶向关系，并采用双荧光素酶报告实验进行验证。ELISA检测细胞中TTR水平，RT-PCR检测miR-223-3p水平，CCK-8法检测细胞增殖，血管生成实验分析血管生成率，Western blot检测血管内皮生长因子（VEGF）、TTR、STAT4和FBXW7蛋白表达。结果　与对照组相比，HG组hRECs中TTR水平升高（P<0.05）。培养24 h时，与HG组相比，HG+TTR组细胞活力明显下降（P<0.05）。与HG组血管生成率（38.12±4.91）%相比，HG+TTR组hRECs血管生成率［（19.46±2.12）%］明显较小（P<0.05）。 与HG+NC mimic组相比，HG+miR-223-3p mimic组hRECs中miR-223-3p表达上调，细胞活力增加（均为P<0.05);与HG+miR-223-3p mimic组相比，HG+TTR+miR-223-3p mimic组hRECs中miR-223-3p表达下调，细胞活力下降（均为P<0.05）。与HG+NC mimic组血管生成率（40.11±4.10）%相比，HG+ miR-223-3p mimic组hRECs血管生成率［（61.52±6.25）%］增多（P<0.05）；与HG+ miR-223-3p mimic组相比，HG+TTR+miR-223-3p mimic组hRECs血管生成率［（42.24±4.33）%］减少（P<0.05），且与HG+NC mimic组比较差异无统计学意义（P>0.05）。生物信息学分析结果显示，STAT4与miR-223-3p启动子区域相互作用。与HG组相比，HG +TTR组hRECs中miR-223-3p、STAT4蛋白表达均降低，FBXW7和VEGF蛋白表达均升高（均为P<0.05）；与HG+TTR组相比，HG+miR-223-3p mimic组hRECs中miR-223-3p、STAT4蛋白表达均升高，FBXW7和VEGF蛋白表达均降低（均为P<0.05）；与HG+miR-223-3p mimic组相比，HG+TTR+miR-223-3p mimic组hRECs中miR-223-3p、STAT4蛋白表达均降低，FBXW7和VEGF蛋白表达均升高（均为P<0.05）；与HG+TTR组相比，HG+TTR+miR-223-3p mimic组hRECs中miR-223-3p、VEGF、STAT4和FBXW7蛋白表达差异均无统计学意义（均为P>0.05）。结论　TTR可能通过调节STAT4/miR-223-3p/FBXW7通路抑制HG诱导的hRECs的血管生成。
Abstract:
Objective　To analyze the effects of the transthyretin (TTR) on high glucose (HG)-induced angiogenesis of human retinal endothelial cells (hRECs) by mediating the pathway of the signal transducer and activator of transcription 4 (STAT4)/miR-223-3p/F-box and WD repeat domain containing 7 (FBXW7). Methods　The hRECs were divided into the control group, HG group, HG+TTR group, HG+ negative control (NC) mimic group, HG+miR-223-3p mimic group and HG+TTR+miR-223-3p mimic group. The targeted relationship between STAT4 and miR-223-3p was analyzed via bioinformatics, which was verified by dual-luciferase reporter assay. The TTR level in cells was detected by the enzyme-linked immunosorbent assay. The miR-223-3p level was detected by real-time fluorescence quantitative PCR. Cell proliferation was detected through the Cell Counting Kit-8. The rate of angiogenesis was detected via the angiogenesis experiment. The expression of vascular endothelial growth factor (VEGF), TTR, STAT4 and FBXW7 was detected via Western blot. Results　Compared with the control group, the TTR level in hRECs increased in the HG group (P<0.05). After 24-hour incubation, compared with the HG group, cell activity significantly decreased in the HG+TTR group (P<0.05). Compared with the HG group ［(38.12±4.91)%］, the rate of angiogenesis was significantly lower in the HG+TTR group ［(19.46±2.12)%］ (P<0.05). Compared with the HG+NC mimic group, expression of miR-223-3p was up-regulated, and the cell activity increased in the HG+miR-223-3p mimic group (both P<0.05). Compared with the HG+miR-223-3p mimic group, expression of miR-223-3p was down-regulated, and cell activity decreased in the HG+TTR+miR-223-3p mimic group (both P<0.05). Compared with the HG+NC mimic group ［(40.11±4.10)%］, the rate of angiogenesis increased in the HG+miR-223-3p mimic group ［(61.52±6.25)%］ (P<0.05). The rate of angiogenesis decreased in the HG+TTR+miR-223-3p mimic group ［(42.24±4.33)%］ (P<0.05) compared with the HG+miR-223-3p mimic group, and had no significant difference from that in the HG+NC mimic group (P>0.05). Bioinformatics analysis results showed an interaction between STAT4 and miR-223-3p promoter region. Compared with the HG group, the expression of miR-223-3p and STAT4 decreased, while the expression of FBXW7 and VEGF increased in the HG+TTR group (all P<0.05). Compared with the HG+TTR group, the expression of miR-223-3p and STAT4 increased, while the expression of FBXW7 and VEGF decreased in the HG+miR-223-3p mimic group (all P<0.05). Compared with the HG+miR-223-3p mimic group, the expression of miR-223-3p and STAT4 decreased, while the expression of FBXW7 and VEGF increased in the HG+TTR+miR-223-3p mimic group (all P<0.05). There was no significant difference in the expression of miR-223-3p, VEGF, STAT4 or FBXW7 between the HG+TTR group and the HG+TTR+miR-223-3p mimic group (all P>0.05). Conclusion　TTR may inhibit HG-induced angiogenesis in hRECs via regulating the STAT4/miR 223 3p/FBXW7 pathway.

参考文献/References:

［1］　AZAD R,SINHA S,NISHANT P.Asymmetric diabetic retino-pathy［J］.Indian J Ophthalmol,2021,69(11):3026-3034.
［2］　JAMPOL L M,GLASSMAN A R,SUN J.Evaluation and care of patients with diabetic retinopathy［J］.N Engl J Med,2020,382(17):1629-1637.
［3］　LIN K Y,HSIH W H,LIN Y B,WEN C Y,CHANG T J.Update in the epidemiology,risk factors,screening,and treatment of diabetic retinopathy［J］.J Diabetes Investig,2021,12(8):1322-1325.
［4］　BEHERA U C,BRAR A S.Socioeconomic status and diabetic retinopathy in India［J］.Indian J Ophthalmol,2021,69(11):2939-2943.
［5］　GILLMORE J D,GANE E,TAUBEL J,KAO J,FONTANA M,MAITLAND M L,et al.CRISPR-Cas9 in vivo gene editing for transthyretin amyloidosis［J］.N Engl J Med,2021,385(6):493-502.
［6］　SUN W,SHI Y,YANG J,SONG X,ZHANG Y,ZHANG W,et al.Transthyretin and retinol-binding protein as discriminators of diabetic retinopathy in type 1 diabetes mellitus［J］.Int Ophthalmol,2022,42(4):1041-1049.
［7］　齐新刚,顾永欣,肖云,杜雷.转甲状腺素蛋白对人视网膜色素上皮细胞增殖、迁移和血管生成的影响机制研究［J］.华南国防医学杂志,2020,34(11):757-761,776.
QI X G,GU Y X,XIAO Y,DU L.Study on the effect mechanism of transthyretin on the proliferation,migration and angiogenesis of human retinal pigment epithelial cells 19 ［J］.Mil Med J South Chin,2020,34(11):757-761,776.
［8］　UEKI K,SASAKO T,OKAZAKI Y,MIYAKE K,NANGAKU M,OHASHI Y,et al.Multifactorial intervention has a significant effect on diabetic kidney disease in patients with type 2 diabetes［J］.Kidney Int,2021,99(1):256-266.
［9］　YADAV J D,OTHEE H,CHAN K A,MAN D C,BELLIVEAU P P,TOWLE J.Transthyretin amyloid cardiomyopathy-current and future therapies［J］.Ann Pharmacother,2021,55(12):1502-1514.
［10］　RAPEZZI C,ELLIOTT P,DAMY T,NATIVI-NICOLAU J,BERK J L,VELAZQUEZ E J,et al.Efficacy of tafamidis in patients with hereditary and wild-type transthyretin amyloid cardiomyopathy: further analyses from attract［J］.JACC Heart Fail,2021,9(2):115-123.
［11］　秦莹,孙筱璐,王东.野生型转甲状腺素蛋白心脏淀粉样变一例［J］.中华心血管病杂志,2021,49(10):1023-1026.
QIN Y,SUN X L,WANG D.A case of wild-type transthyretin cardiac amyloidosis ［J］.Chin J Cardiol,2021,49(10):1023-1026.
［12］　胡晓丽.转甲状腺素蛋白与2型糖尿病、糖调节受损的关联性研究［D］.华中科技大学,2019.
HU X L.Correlation between transthyretin and type 2 diabetes mellitus,impaired glucose regulation ［D］.Huazhong Univ Sci Technol,2019.
［13］　JUCHNICKA I,KUZMICKI M.Influence of miRNAs in gestational diabetes mellitus development［J］.Ginekol Pol,2021,92(8):579-582.
［14］　POURHANIFEH M H,MEHRZADI S,HOSSEINZADEH A.Melatonin and regulation of miRNAs: novel targeted therapy for cancerous and noncancerous disease［J］.Epigenomics,2021,13(1):65-81.
［15］　JIANG Q,LIU C,LI C P,XU S S,YAO M D,GE H M,et al.Circular RNA-ZNF532 regulates diabetes-induced retinal pericyte degeneration and vascular dysfunction［J］.J Clin Invest,2020,130(7):3833-3847.
［16］　林莹,黎绮雯,林小媚.miR-15和miR-29a在糖尿病视网膜病变患者血清中的表达及临床意义［J］.中国医师杂志,2020,22(3):398-402.
LIN Y,LI Q W,LIN X M.Expressions and clinical significances of miR-15 and miR-29a in serum of patients with diabetic retinopathy ［J］.J Chin Physician,2020,22(3):398-402.
［17］　IZUMIYA Y,HAYASHI H,ISHIKAWA H,SHIBATA A,YOSHIYAMA M.How to identify transthyretin cardiac amyloidosis at an early stage［J］.Intern Med,2021,60(1):1-7.
［18］　YIN Z,TAN R,YUAN T,CHEN S,QUAN Y.Berberine prevents diabetic retinopathy through inhibiting HIF-1α /VEGF/ NF-κB pathway in db/db mice［J］.Pharmazie,2021,76(4):165-171.
［19］　WANG L,YANG J,XIAO X,ZHENG C,MING D.VEGF modulates the neural dynamics of hippocampal subregions in chronic global cerebral ischemia rats［J］.Neuromolecular Med,2021,23(3):416-427.
［20］　NIE X,JIA W,LI X,PAN X,YIN R,LIU N,et al.FBXW7 induces apoptosis in glioblastoma cells by regulating HDAC7［J］.Cell Biol Int,2021,45(10):2150-2158.
［21］　WANG Y,SHI S,WANG Y,ZHANG X,LIU X,LI J,et al.miR-223-3p targets FBXW7 to promote epithelial-mesenchymal transition and metastasis in breast cancer［J］.Thorac Cancer,2022,13(3):474-482.

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备注/Memo

备注/Memo:
河南省医学科技攻关计划（联合共建）项目（编号：LHGJ20190492）
更新日期/Last Update: 2023-02-05