糖尿病诱导硫氧还蛋白相互作用蛋白(TXNIP)对小鼠胰岛β细胞衰老的影响*

doi:10.12047/j.cjap.5878.2020.027

摘要/Abstract

摘要： 目的: 本研究旨在观察糖尿病中硫氧还蛋白相互作用蛋白(TXNIP)的表达是否影响胰岛β细胞衰老。方法: 正常小鼠(db/m)、糖尿病小鼠(db/db)随机各取6只,血糖仪检测其空腹血糖值,Western blot检测胰腺组织TXNIP蛋白表达、免疫化学染色检测胰腺组织中衰老相关β-半乳糖苷酶活性,Western blot检测胰腺组织衰老相关指标p16、p21、Rb表达的变化。INS-1胰岛β细胞随机分7组(n=6),用各组慢病毒(30 μl)转染4~6 h后,嘌呤霉素(PM, 3 μg/m)筛选7 d,构建Normal组(正常组)、Scramble ShRNA组(干扰空病毒组)、TXNIP-ShRNA-1(TXNIP沉默一组)组、TXNIP-ShRNA-2(TXNIP沉默二组)组、TXNIP-ShRNA-3组(TXNIP沉默三组)、Ad-GFP组(过表达空病毒组)、Ad-TXNIP-GFP组(TXNIP过表达组)稳转INS-1胰岛β细胞株,检测其TXNIP蛋白表达、衰老相关β -半乳糖苷酶活性、衰老相关指标。结果: 与正常小鼠相比,db/db组小鼠空腹血糖显著上升(P＜0.01),胰腺组织TXNIP蛋白表达显著升高(P＜0.05),胰腺组织β -半乳糖苷酶阳性染色率增加,p16、p21、Rb蛋白表达显著升高(P＜ 0.05)。与Ad-GFP组相比,Ad-TXNIP-GFP组β -半乳糖苷酶阳性染色率增加,p16、p21、Rb蛋白表达均显著增加(P＜0.01)。与Scramble ShRNA组相比,TXNIP-ShRNA组β -半乳糖苷酶阳性染色率降低,p16、p21以及Rb蛋白表达均降低(P＜0.05)。结论: 糖尿病可通过上调TXNIP表达,诱导胰岛β细胞衰老。

关键词: 糖尿病, 硫氧还蛋白相互作用蛋白, INS-1胰岛β细胞, 细胞衰老, 小鼠

Abstract: Objective: To investigate whether the increased expression of thioredoxin interacting protein (TXNIP) in diabetes affects the senescence of islet β cells. Methods: Six normal mice (db/m) and six diabetic mice (db/db) were randomly selected. Fasting blood glucose was measured by blood sugar meter, the expression levels of TXNIP protein, p16, p21 and Rb in pancreatic tissues were detected by Western blot, senescence-associated beta-galactosidase activity in pancreatic tissue was determined by immunochemical staining. INS-1 islet beta cells were randomly divided into 7 groups (n=6), and transfected with lentiviruses (30 μl) for 4 to 6 hours, then was screened with puromycin (PM, 3 μg/m) for 7 days to construct normal group, scramble ShRNA group (interference with airborne poison group), TXNIP-ShRNA-1 group (TXNIP silence group-1), TXNIP-ShRNA-2 group (TXNIP silence group 2), TXNIP-ShRNA-3 group (TXNIP silence group 3), Ad-GFP group (overexpression of the air virus group), Ad-TXNIP-GFP group (TXNIP overexpression group) stably transferred INS-1 islet beta cell line. TXNIP protein expression was detected by Western blot, aging-related beta-galactosidase activity was detected by immunochemical staining, the changes of expression of p16, p21 and Rb was determined by Western blot. Results: Compared with normal mice, the fasting blood glucose of db/db group was increased significantly (P＜0. 01), the expression of TXNIP protein was increased significantly in pancreatic tissues(P＜0. 05), positive staining rate of β- galactosidase was increased significantly in pancreatic tissues, p16/p21/Rb protein expression levels were increased significantly (P＜0. 05). Compared with Ad-GFP group, the positive staining rate of β- galactosidase in Ad-TXNIP-GFP group was increased significantly, p16/p21/Rb protein expression levels were increased significantly (P＜0. 01). Compared to the scramble ShRNA group, the positive staining rate of β- galactosidase in TXNIP-ShRNA group was decreased, p16/p21/Rb protein expression levels were decreased significantly (P＜0. 05). Conclusion: Diabetes can induce islet β-cell senescence by up-regulating TXNIP expression.

Key words: diabetes, thioredoxin interacting protein, INS-1 cell, cell senescence

中图分类号:

R338

霍海燕, 王瑾, 张许梅, 张文婷, 岳继萍, 焦向英. 糖尿病诱导硫氧还蛋白相互作用蛋白(TXNIP)对小鼠胰岛β细胞衰老的影响^*[J]. 中国应用生理学杂志, 2020, 36(2): 119-123.

HUO Hai-yan, WANG Jin, ZHANG Xu-mei, ZHANG Wen-ting, YUE Ji-ping, JIAO Xiang-ying. Effect of diabetic induced thioredoxin interacting protein (TXNIP) on islet cell senescence[J]. CJAP, 2020, 36(2): 119-123.

参考文献

[1] Ogurtsova K, da Rocha Fernandes JD, Huang Y, et al. IDF diabetes atlas: global estimates for the prevalence of diabetes for 2015 and 2040[J]. Diabetes Res Clin Pract, 2017, 128: 40-50.
[2] 王雪, 杨静, 雷燕. 衰老相关分泌表型与血管衰老的研究进展[J]. 中华老年医学杂志, 2017, 36(10)∶1151-1155.
[3] Yosef R, Pilpel N, Tokarsky-Amiel R, et al. Directed elimination of senescent cells by inhibition of BCL-W and BCL-XL[J]. Nat Commun, 2016, 7∶11190.
[4] Palmer AK, Tamara T, Lebrasseur NK, et al. Cellular senescence in type 2 diabetes∶ A therapeutic opportunity[J]. Diabetes, 2015, 64(7)∶ 2289-2298.
[5] Höhn A, Weber D, Jung T, et al. Happily (n)ever after∶ Aging in the context of oxidative stress, proteostasis loss and cellular senescence[J]. Redox Biol, 2017, 11 (C)∶ 482-501.
[6] Junn E, Han SH, Im JY, et al. Vitamin D3 up-regulated protein 1 mediates oxidative stress via suppressing the thioredoxin function[J]. J Immunol, 2000, 164(12)∶ 6287-6295.
[7] 李杨. TXNIP在衰老导致的胰岛β细胞功能障碍中的作用及其机制研究 [D]. 重庆医科大学, 2017.
[8] 项伟玲, 金丽琴, 高峰, 等. 黑苦荞茎叶对2型糖尿病小鼠的治疗作用及其对其胰腺、脾脏的影响[J]. 中国应用生理学杂志, 2019, 35(2)∶ 140-144.
[9] Ji L, Wang Q, Huang F, et al. FOXO1 overexpression attenuates tubulointerstitial fibrosis and apoptosis in diabetic kidneys by ameliorating oxidative injury via TXNIP-TRX[J]. Oxid Med Cell Longev, 2019: 6431359.
[10]曹竹杰, 冯艳金, 李丹, 等. 腺病毒过表达TXNIP抑制INS-1胰岛β细胞增殖[J]. 生理学报, 2018, 70(2)∶ 158-166.
[11]Riahi Y, Kaiser N, Cohen G, et al. Foam cell-derived 4-hydroxynonenal induces endothelial cell senescence in a TXNIP-dependent manner[J]. J Cell Mol Med, 2015, 19(8)∶1887-1899.
[12]赵传祥, 李宏慧, 汤思熠, 等. 间充质干细胞培养上清液抑制成纤维细胞的衰老[J]. 江苏大学学报(医学版), 2019, 29(1)∶ 17-21.
[13]Alsayegh KN, Gadepalli VS, Iyer S, et al. Knockdown of CDK2AP1 in primary human fibroblasts induces p53 dependent senescence[J]. Plos One, 2015, 10(3)∶ e0120782.
[14]邹美圣, 刘凌, 刘泽, 等. 灵芝多糖对血管紧张素Ⅱ诱导的血管内皮细胞衰老的干预研究[J]. 现代中西医结合杂志, 2012, 21(13)∶ 1386-1387.
[15]孙伟铭, 张源洲, 温馨, 等. 外源性H₂S恢复缺氧后适应对衰老H9C2细胞的保护作用及机制[J]. 中国应用生理学杂志, 2018, 34(4)∶ 3-7.

糖尿病诱导硫氧还蛋白相互作用蛋白(TXNIP)对小鼠胰岛β细胞衰老的影响^*

Effect of diabetic induced thioredoxin interacting protein (TXNIP) on islet cell senescence

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