电针通过调节IκB激酶β表达抑制局灶性脑缺血再灌注大鼠脑内炎症损害的机制

doi:10.3969/j.issn.1006-9771.2019.04.008

《中国康复理论与实践》 ›› 2019, Vol. 25 ›› Issue (4): 407-415.doi: 10.3969/j.issn.1006-9771.2019.04.008

电针通过调节IκB激酶β表达抑制局灶性脑缺血再灌注大鼠脑内炎症损害的机制

秦文熠¹, 荣晓凤¹, 罗勇²

1.重庆医科大学附属第一医院中西医结合科；重庆市 400016
2.重庆医科大学附属第一医院神经内科，重庆市 400016

收稿日期:2018-08-25 修回日期:2019-01-11 出版日期:2019-04-20 发布日期:2019-04-24
通讯作者: 罗勇，E-mail: Luoyong1998@163.com
作者简介:秦文熠(1983-)，女，汉族，江苏海门市人，博士研究生，主治医师，主要研究方向：脑缺血再灌注损伤与修复机制。
基金资助:
1.国家自然科学基金青年基金项目(No. 81403243)；2.重庆市卫计委中医药科技项目(No. ZY201702073)

Effect of Electroacupuncture on Inflammatory Response of Rats after Focal Cerebral Ischemia-reperfusion via Regulating IκB Kinases β Expression

QIN Wen-yi¹, RONG Xiao-feng¹, LUO Yong²

1.the Integrated Traditional Chinese and Western Medicine Department， the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
2.Department of Neurology, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China

Received:2018-08-25 Revised:2019-01-11 Published:2019-04-20 Online:2019-04-24
Contact: LUO Yong, E-mail: Luoyong1998@163.com
Supported by:
National Natural Science Foundation of China (No. 81403243) and the Program of The Traditional Chinese Medicine Research of Chongqing Municipal Health and Planning Commission (No. ZY201702073)

摘要/Abstract

摘要： 目的观察局灶脑缺血再灌注后大鼠脑内炎性损害的变化，探讨炎症反应中激活核因子-κB (NF-κB)信号通路的关键蛋白IκB激酶(IKK)β的启动作用及电针抑制炎性损害的机制。方法将雄性Sprague-Dawley大鼠240只按随机数字表法随机分为假手术组、模型组、电针组、IKKβ沉默组、IKKβ过表达组和IKKβ过表达+电针组，每组设再灌注后6 h、12 h、24 h、48 h和72 h共5个时间点。利用改良线栓法制备右侧大脑中动脉闭塞再灌注模型。利用基因沉默及基因过表达技术对IKKβ基因进行干预。结果与模型组比较，IKKβ沉默组神经功能评分升高(P < 0.05)，脑梗死体积减小(P < 0.05)，NF-κB p65激活受抑制，促炎因子含量降低(P < 0.05)。与IKKβ沉默组比较，IKKβ过表达组上述结果均明显变差(P < 0.05)，且大脑缺血皮质区小胶质细胞明显活化。IKKβ过表达+电针组小胶质细胞活化及IKKβ激活明显受抑制。结论 IKKβ基因沉默能明显抑制NF-κB信号通路介导的大脑缺血皮质区的炎症反应，IKKβ过表达则缺血皮质区炎性损害程度较重。电针通过调节IKKβ活性从而抑制局灶脑缺血再灌注后的炎症损害。

关键词: 局灶脑缺血再灌注, 电针, 炎症反应, IκB激酶β, 基因干扰, 小胶质细胞, 大鼠

Abstract: Objective To observe the changes of inflammatory damage in the brain of rats after focal cerebral ischemia-reperfusion, and explore the effect of the initiation of IκB kinases β (IKKβ), which is the key protein of activating nuclear factor (NF)-kappa B signaling pathway in inflammatory response, and the mechanism of electroacupuncture inhibiting inflammatory damage. Methods A total of 240 male Sprague-Dawley rats were randomly divided into sham group, ischemia-reperfusion group, electroacupuncture group, IKKβ silencing group, IKKβ overexpression group and IKKβ overexpression + electroacupuncture group, each group was further divided into six hours, twelve hours, 24 hours, 48 hours and 72 hours subgroups. The right middle cerebral artery occlusion reperfusion model was established by modified thread embolization. The IKKβ gene was intervened by gene silencing and gene overexpression technology. Results Compared with the model group, the neurological function score increased (P < 0.05), the cerebral infarction volume decreased (P < 0.05), the activation of NF-κB p65 was inhibited, and the content of proinflammatory factors decreased (P < 0.05) in IKKβ silencing group. Compared with IKKβ silencing group, the above results were significantly worse in IKKβ overexpression group (P < 0.05), and microglia in cerebral ischemic cortex were significantly activated. The activation of microglia and activation of IKKβ were significantly inhibited in IKKβ overexpression + electroacupuncture group. Conclusion IKKβ gene silencing could inhibit the inflammatory response of cerebral ischemic cortex mediated by NF-κB signaling pathway, and over-expression of IKKβ could lead to severe inflammatory damage in ischemic cortex. Electroacupuncture could inhibit the inflammatory damage after focal cerebral ischemia-reperfusion by regulating the activity of IKKβ.

Key words: focal cerebral ischemia-reperfusion, electroacupuncture, inflammation, IκB kinases β, gene silence, microglial cell, rats

中图分类号:

R743.3

秦文熠, 荣晓凤, 罗勇. 电针通过调节IκB激酶β表达抑制局灶性脑缺血再灌注大鼠脑内炎症损害的机制[J]. 《中国康复理论与实践》, 2019, 25(4): 407-415.

QIN Wen-yi, RONG Xiao-feng, LUO Yong. Effect of Electroacupuncture on Inflammatory Response of Rats after Focal Cerebral Ischemia-reperfusion via Regulating IκB Kinases β Expression[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2019, 25(4): 407-415.

参考文献

[1] XuH, MuS, QinW. Microglia TREM2 is required for electroacupuncture to attenuate neuroinflammation in focal cerebral ischemia/reperfusion rats [J]. Biochem Biophys Res Commun, 2018, 503(4): 3225-3234.
[2] XuH, QinW, HuX, et al. Lentivirus-mediated overexpression of OTULIN ameliorates microglia activation and neuroinflammation by depressing the activation of the NF-κB signaling pathway in cerebral ischemia/reperfusion rats [J]. J Neuroinflammation, 2018, 15(1): 83-96.
[3] AkbariG, Ali MardS, VeisiA. A comprehensive review on regulatory effects of crocin on ischemia/reperfusion injury in multiple organs [J]. Biomed Pharmacother, 2018, 99: 664-670.
[4] IadecolaC, AnratherJ. The immunology of stroke: from mechanisms to translation [J]. Nat Med, 2011, 17(7): 796-808.
[5] JiB, ChengB, PanY, et al. Neuroprotection of bradykinin/bradykinin B2 receptor system in cerebral ischemia [J]. Biomed Pharmacother, 2017, 94: 1057-1063.
[6] LambertsenK L, BiberK, FinsenB. Inflammatory cytokines in experimental and human stroke [J]. J Cereb Blood Flow Metab, 2012, 32(9): 1677-1698.
[7] LatanichC A, Toledo-PereyraL H. Searching for NF-kappaB-based treatments of ischemia reperfusion injury [J]. J Invest Surg, 2009, 22(4): 301-315.
[8] LiuX, ZhangX, WangF, et al. Improvement in cerebral ischemia-reperfusion injury through the TLR4/NF-κB pathway after Kudiezi injection in rats [J]. Life Sci, 2017, 191: 132-140.
[9] SuD, ChengY, LiS, et al. Sphk1 mediates neuroinflammation and neuronal injury via TRAF2/NF-κB pathways in activated microglia in cerebral ischemia reperfusion [J]. J Neuroimmunol, 2017, 305: 35-41.
[10] BrambillaR, Bracchi-RicardV, HuWH, et al. Inhibition of astroglial nuclear factor kappaB reduces inflammation and improves functional recovery after spinal cord injury [J]. J Exp Med, 2005, 202(1): 145-156.
[11] HuX, LiP, GuoY, et al. Microglia/macrophage polarization dynamics reveal novel mechanism of injury expansion after focal cerebral ischemia [J]. Stroke, 2012, 43(11): 3063-3070.
[12] MiyamotoS. Nuclear initiated NF-κB signaling: NEMO and ATM take center stage [J]. Cell Res, 2011, 21(1): 116-130.
[13] OritaS, MiyagiM, KoboriS, et al. IκB kinase β inhibitor downregulates pain-related neuropeptide production in the sensory neurons innervating injured lumbar intervertebral discs in the dorsal root ganglia of rats [J]. Spine J, 2013, 13(3): 284-288.
[14] SamaniegoS, MarcuK B. IKKβ in myeloid cells controls the host response to lethal and sublethal Francisella tularensis LVS infection [J]. PLoS One, 2013, 8(1): e54124.
[15] ZhangS, QiY, XuY, et al. Protective effect of flavonoid-rich extract from Rosa laevigata Michx on cerebral ischemia-reperfusion injury through suppression of apoptosis and inflammation [J]. Neurochem Int, 2013, 63(5): 522-532.
[16] ZhanJ, QinW, ZhangY, et al. Upregulation of neuronal zinc finger protein A20 expression is required for electroacupuncture to attenuate the cerebral inflammatory injury mediated by the nuclear factor-κB signaling pathway in cerebral ischemia/reperfusion rats [J]. J Neuroinflammation, 2016, 13(1): 258-262.
[17] ZengY J, TsaiS Y, ChenK B, et al. Comparison of electroacupuncture and morphine-mediated analgesic patterns in a plantar incision-induced pain model [J]. Evid Based Complement Alternat Med, 2014, 2014: 659343.
[18] GuoF, SongW, JiangT, et al. Electroacupuncture pretreatment inhibits NADPH oxidase-mediated oxidative stress in diabetic mice with cerebral ischemia [J]. Brain Res, 2014, 1573: 84-91.
[19] ChoiJ W, KangS Y, ChoiJ G, et al. Analgesic effect of electroacupuncture on paclitaxel-induced neuropathic pain via spinal opioidergic and adrenergic mechanisms in mice [J]. Am J Chin Med, 2015, 43(1): 57-70.
[20] DuY, ShiL, LiJ, et al. Angiogenesis and improved cerebral blood flow in the ischemic boundary area were detected after electroacupuncture treatment to rats with ischemic stroke [J]. Neurol Res, 2011, 33(1): 101-107.
[21] LiuY, ZouL P, DuJ B, et al. Electro-acupuncture protects against hypoxic-ischemic brain-damaged immature rat via hydrogen sulfide as a possible mediator [J]. Neurosci Lett, 2010, 485(1): 74-78.
[22] YuJ B, ShiJ, GongL R, et al. Role of Nrf2/ARE pathway in protective effect of electroacupuncture against endotoxic shock-induced acute lung injury in rabbits [J]. PLoS One, 2014, 9(8): 705-712.
[23] LiuA J, LiJ H, LiH Q, et al. Electroacupuncture for acute ischemic stroke: a meta-analysis of randomized controlled trials [J]. Am J Chin Med, 2015, 43(8): 1541-1566.
[24] LiX, LuoP, WangQ, et al. Electroacupuncture pretreatment as a novel avenue to protect brain against ischemia and reperfusion injury [J]. Evid Based Complement Alternat Med, 2012, 2012(5): 195397.
[25] QinW Y, LuoY, ChenL, et al. Electroacupuncture could regulate the NF-κB signaling pathway to ameliorate the inflammatory injury in focal cerebral ischemia/reperfusion model rats [J]. Evid Based Complement Alternat Med, 2013, 2013: 924541.
[26] LongaE Z, WeinsteinP R, CarlsonS, et al. Reversible middle cerebral artery occlusion without craniectomy in rats [J]. Stroke, 1989, 20(1): 84-91.
[27] JiangJ, LuoY, QinW, et al. Electroacupuncture suppresses the NF-κB signaling pathway by upregulating cylindromatosis to alleviate inflammatory injury in cerebral ischemia/reperfusion rats [J]. Front Mol Neurosci, 2017, 10: 363-369.
[28] 秦文熠,罗勇,余超. 电针对局灶性脑缺血再灌注大鼠海马内白介素-1β及转录核因子κB抑制蛋白激酶的影响[J]. 针刺研究, 2013, 38(4): 271-276.
[29] 张莹,秦文熠,罗勇. 电针对局灶脑缺血/再灌注大鼠大脑缺血皮质区白介素-4/信号传导及转录激活因子6与核因子-κB的影响[J]. 针刺研究, 2017, 42(3): 217-222, 258.
[30] GarciaJ H, WagnerS, LiuK F, et al. Neurological deficit and extent of neuronal necrosis attributable to middle cerebral artery occlusion in rats. Statistical validation [J]. Stroke, 1995, 26(4): 627-634, discussion 635.
[31] 秦文熠,荣晓凤,陶涛,等. 核转录因子-κB拮抗剂对大鼠海马CA1区局灶性脑缺血再灌注损伤的影响[J]. 中华神经科杂志, 2016, 49(1): 45-53.
[32] JinR, YangG, LiG. Inflammatory mechanisms in ischemic stroke: role of inflammatory cells [J]. J Leukoc Biol, 2010, 87(5): 779-789.
[33] NimmerjahnA, KirchhoffF, HelmchenF. Resting microglial cells are highly dynamic surveillants of brain parenchyma in vivo [J]. Science, 2005, 308(5726): 1314-1318.
[34] M èTremblay, StevensB, SierraA, et al. The role of microglia in the healthy brain [J]. J Neurosci, 2011, 31(45): 16064-16069.
[35] TuX K, YangW Z, ShiS S, et al. Spatio-temporal distribution of inflammatory reaction and expression of TLR2/4 signaling pathway in rat brain following permanent focal cerebral ischemia [J]. Neurochem Res, 2010, 35(8): 1147-1155.
[36] KettenmannH, VerkhratskyA. Neuroglia: the 150 years after [J]. Trends Neurosci, 2008, 31: 653-659.
[37] SimmonsL J, Surles-ZeiglerM C, LiY, et al. Regulation of inflammatory responses by neuregulin-1 in brain ischemia and microglial cells in vitro involves the NF-kappa B pathway [J]. J Neuroinflammation, 2016, 13: 237-242.

电针通过调节IκB激酶β表达抑制局灶性脑缺血再灌注大鼠脑内炎症损害的机制

Effect of Electroacupuncture on Inflammatory Response of Rats after Focal Cerebral Ischemia-reperfusion via Regulating IκB Kinases β Expression

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