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山东大学学报（医学版） ›› 2017, Vol. 55 ›› Issue (3): 6-11.doi: 10.6040/j.issn.1671-7554.0.2016.1591

• 基础医学 • 上一篇    下一篇

Nod样受体蛋白3炎性体在2型糖尿病脑微血管内皮细胞中的变化及变化机制

李帅1,王雅琳2,孙忠文3,朱梅佳4   

  1. 1. 山东大学医学院, 山东 济南 250012;2.首都医科大学肿瘤研究所, 北京 100069;3.烟台毓璜顶医院神经内科, 山东 烟台 264000;4.山东大学附属千佛山医院神经内科, 山东 济南 250014
  • 收稿日期:2016-11-30 出版日期:2017-03-10 发布日期:2017-03-10
  • 通讯作者: 朱梅佳. E-mail: zhumeijia2014@163.com E-mail:zhumeijia2014@163.com
  • 基金资助:
    山东省自然科学基金(ZR2010L1M101)

Expression changes and mechanism of Nod-like receptor protein 3 inflammasome in the cerebral microvascular endothelial cells in type 2 diabetes mellitus

LI Shuai1, WANG Yalin2, SUN Zhongwen3, ZHU Meijia4   

  1. 1. School of Medicine, Shandong University, Jinan 250012, Shandong, China;
    2. Tumor Research Institute, Capital Medical University, Beijing 100069, China;
    3. Department of Neurology, Yantai Yuhuangding Hospital, Yantai 264000, Shandong, China;
    4. Department of Neurology, Qianfoshan Hospital of Shandong University, Jinan 250012, Shandong, China
  • Received:2016-11-30 Online:2017-03-10 Published:2017-03-10

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摘要： 目的 探讨Nod样受体蛋白3(NLRP3)炎性体在2型糖尿病脑微血管内皮中的变化及机制。 方法 3月龄Wistar大鼠和自发性2型糖尿病(GK)大鼠各5只,采用免疫组织化学法观察NLRP3在脑组织中的表达;体外培养小鼠脑微血管内皮细胞(CMEC)。不同糖浓度培养基模拟2型糖尿病内环境,活性氧(ROS)阻断剂N-乙酰半胱氨酸(NAC)作为抑制剂,将细胞分为对照组(糖浓度5.6 mmol/L)、高糖1组(HG1组,培养基糖浓度10 mmol/L)、高糖2组(HG2组,培养基糖浓度20 mmol/L)、高糖3组(HG3,培养基糖浓度30 mmol/L)及高糖+NAC组(HG+NAC组,目的蛋白表达较明显组的糖浓度)。采用Western blotting法检测各组硫氧还蛋白交互蛋白(TXNIP)和NLRP3的表达,并对TXNIP和NLRP3是否存在相关性进行分析;采用ELISA法检测白介素1β(IL-1β)的含量;采用流式细胞术检测对照组、HG3组、HG+NAC组ROS的含量。 结果 NLRP3在脑微血管壁聚集,与Wistar大鼠相比,GK大鼠的阳染强度,阳染血管数均有增加(P<0.05);与对照组相比,HG1组、HG2组、HG3组TXNIP、NLRP3的表达增加(P<0.01),HG3组最明显,细胞内IL-1β水平增高(P<0.01),ROS的含量增加(P<0.01);细胞内TXNIP和NLRP3的表达呈正相关性(r=0.993;P<0.05);给予ROS清除剂后,与HG3组相比,HG+NAC组细胞内ROS含量下降(P<0.01),TXNIP、NLRP3表达水平下降(P<0.01),细胞内IL-1β水平下降(P<0.01)。 结论 NLRP3在2型糖尿病脑微血管内皮细胞中经ROS-TXNIP途径被激活,并且促进炎性因子IL-β的释放,在2型糖尿病脑微血管损伤中发挥重要作用。

关键词: Nod样受体蛋白3炎性体, 2型糖尿病, 脑微血管内皮细胞, 硫氧还蛋白交互蛋白, 活性氧

Abstract: Objective To investigate the expression changes and mechanism of Nod-like receptor protein 3(NLRP3)inflammasome in cerebral microvascular endothelial cells of type2 diabetes mellitus. Methods A total of 3-month old Wistar rats(n=5)and spontaneous type 2 diabetic rats(GK, n=5)were enrolled. The expression of NLRP3 in the brain tissues was observed with immunohistochemical method. After cerebral microvascular endothelial cells(CMECs)were cultivated in vitro, they were divided into 5 groups: normal control group, high glucose group 1(HG1 group, glucose concentration=10 mmol/L), high glucose group 2(HG2 group, glucose concentration=20 mmol/L), high 山 东 大 学 学 报 (医 学 版)55卷3期 -李帅,等.Nod样受体蛋白3炎性体在2型糖尿病脑微血管内皮细胞中的变化及变化机制 \=-glucose group 3(HG3 group, glucose concentration=30 mmol/L), and HG+NAC group. The protein expressions of thioredoxin interacting protein(TXNIP)and NLRP3 were detected with Western blotting. The relationship between NLRP3 and TXNIP was analyzed. The content of interleukin-1β(IL-1β)was evaluated with ELISA kits. The content of intracellular reactive oxygen species(ROS)was measured with flow cytometry. Results NLRP3 gathered in microvascular endothelial cells. Compared with Wistar rats, GK rats had increased strength and number of vascular with positive staining(P<0.05). Compared with normal control group, HG1, HG2 and HG3 groups had higher expressions of TXNIP and NLRP3(P<0.01), especially in the HG3 group, in which the level of IL-1β and ROS also increased (P<0.01). TXNIP and NLRP3 was positively correlated(r=0.993; P<0.05). Lompared with HG3 group, in the HG+NAC group, the level of intracellular ROS, TXNIP, NLRP3 and IL-1β decreased(P<0.01). Conclusion NLRP3 inflammasome is activated in type 2 diabetes mellitus by ROS-TXNIP pathway. It promotes the release of inflammatory factor IL-1β and plays an important role in the microvascular injury.

Key words: Type 2 diabetes mellitus, Reactive oxygen species, Nod-like receptor protein 3 inflammasome, Cerebral microvascular endothelial cells, Thioredoxin interacting protein

中图分类号: 

  • R58
[1] Prasad S, Naik P, Cucullo L, et al. Diabetes mellitus and blood-brain barrier dysfunction: an overview[J]. J Pharmacovigil, 2014, 2(2): 125.
[2] Sun YN, Liu LB, Xue YX, et al. Effects of insulin combined with idebenone on blood-brain barrier permeability in diabetic rats [J]. J Neurosci Res, 2015, 93(4): 666-677.
[3] Umegaki H, Hayashi T, Nomura H, et al. Cognitive dysfunction: an emerging concept of a new diabetic complication in the elderly [J]. Geriatr Gerontol Int, 2013, 13(1): 28-34.
[4] Saedi E, Gheini MR, Faiz F, et al. Diabetes mellitus and cognitive impairments [J]. World J Diabetes, 2016, 7(17): 412-422.
[5] Price TO, Eranki V, Banks WA, et al. Topiramate treatment protects blood-brain barrier pericytes from hyperglycemia-induced oxidative damage in diabetic mice [J]. Endocrinology, 2012, 153(1): 362-372.
[6] Brownlee M. Biochemistry and molecular cell biology of diabetic complications [J]. Nature, 2001, 13(414): 813-840.
[7] Soares E, Prediger RD, Nunes S, et al. Spatial memory impairments in a prediabetic rat model [J]. Neuroscience, 2013, 250(10): 565-577.
[8] Shah GN, Morofuji Y, Banks WA, et al. High glucose-induced mitochondrial respiration and reactive oxygen species in mouse cerebral pericytes is reversed by pharmacological inhibition of mitochondrial carbonic anhydrases: Implications for cerebral microvascular disease in diabetes [J]. Biochem Biophys Res Commun, 2013, 440(2): 354-358.
[9] Yunpeng D, Casey MM, Kern T, et al. Hyperglycemia increases mitochondrial superoxide in retina and retinal cells [J]. Free Radic Biol Med, 2003, 35(11): 1491-1499.
[10] Wang P, Huang R, Lu S, et al. RAGE and AGEs in mild cognitive impairment of diabetic patients: a cross-sectional study [J]. PLoS One, 2016, 11(1): 0145521. doi:10.1371.
[11] Bauernfeind F, Hornung V. Of inflammasomes and pathogens - sensing of microbes by the inflammasome [J]. EMBO Mol Med, 2013, 5(6): 814-826.
[12] Gombault A, Baron L, Couillin I, et al. ATP release and purinergic signaling in NLRP3 inflammasome activation [J]. Front Immunol, 2012, 3(11): 414.
[13] Lima HJr, Jacobson LS, Goldberg MF, et al. Role of lysosome rupture in controlling Nlrp3 signaling and necrotic cell death [J]. Cell Cycle, 2013, 12(12): 1868-1878.
[14] Lane T, Flam B, Lockey R, et al. TXNIP shuttling: missing link between oxidative stress and inflammasome activation [J]. Front Physiol, 2013, 4: 50.
[15] Chen KH, Zhang JG, Zhang WW, et al. ATP-P2X4 signaling mediates NLRP3 inflammasome activation: a novel pathway of diabetic nephropathy [J]. Int J Biochem Cell Biol, 2013, 45(5): 932-943.
[16] Zhou R, Tardivel A, Thorens B, et al. Thioredoxin-interacting protein links oxidative stress to inflammasome activation [J]. Nat Immunol, 2010, 11(2): 136-140.
[17] Latz E, Xiao TS, Stutz A, et al. Activation and regulation of the inflammasomes [J]. Nat Rev Immunol, 2013, 13(6): 397-411.
[18] Shaked M, Kaiser N, Leibowitz G, et al. Insulin counteracts glucotoxic effects by suppressing thioredoxin-interacting protein production in INS-1E beta cells and in Psammomys obesus pancreatic islets [J]. Diabetologia, 2009, 52(4): 636-644.
[19] Oslowski CM, Hara T O, Sullivan-Murphy B, et al. Thioredoxin-interacting protein mediates ER stress-induced beta cell death through initiation of the inflammasome [J]. Cell Metab, 2012, 16(2): 265-273.
[20] 谢荣辉, 周师洁, 殷明. N-乙酰半胱氨酸对过氧化氢诱导的骨髓间充质干细胞凋亡的保护及作用机制研究[J]. 中国药理学通报, 2014, 30(1): 54-59.
[21] Yang F, Wang Z, Wei X, et al. NLRP3 deficiency ameliorates neurovascular damage in experimental ischemic stroke [J]. J Cereb Blood Flow Metab, 2014, 34(4): 660-667.
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