紫草素对缺血性脑损伤大鼠脑组织保护作用的研究
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摘要
目的探讨紫草素对缺血性脑损伤大鼠脑组织的保护作用及其机制。方法选取48只大鼠,随机分为假手术组、模型组和紫草素治疗组,每组16只。制备缺血性脑损伤大鼠模型,观察3组大鼠日常行为,并对大鼠脑组织进行病理学观察,用酶联免疫吸附试验测定3组脑组织中白介素-6(IL-6)和IL-8含量,蛋白免疫印迹法检测3组脑核因子-κB(NF-κB)蛋白表达水平。结果模型组动物活动减少,部分动物出现躁动。假手术组大鼠皮层及海马区锥体神经元层次分明;模型组皮层及海马区神经细胞层次紊乱,密度降低;紫草素治疗组可以改善缺血性脑损伤大鼠脑组织神经元的异常形态变化。假手术组神经元形态基本正常;模型组神经元细胞胞质内空泡形成,细胞核内染色质边集;紫草素治疗组神经元细胞状态有所好转,细胞变性程度较模型组缓解。紫草素治疗组IL-6和IL-8表达量及NF-κB蛋白表达水平均低于模型组(P<0.01)。结论紫草素对缺血性脑损伤大鼠有一定改善作用。
Objective To investigate protective effect and mechanism of Shikonin on brain tissues of rats ischemic brain injury. Methods A total of 48 rats were randomly divided into sham operation group, model group and Shikonin treatment group(n=16 in each group). Rats models with ischemic brain injury were prepared to observe daily behavior, and pathological observation for brain tissues were performed. Enzyme linked immunosorbent assay was used to detect contents of interleukin-6(IL-6) and interleukin-8(IL-8), and protein expression levels of nuclear factor-κB(NF-κB) were detected by Western blot among three groups. Results In model group, animal activity was decreased, and some animals appeared restlessness; In sham operation group, neurons in cortex and hippocampal cone ordered clearly. In model group, neurons cell layers in cortex and hippocampal cone disordered with decreased density. Shikonin treatment group could improve abnormal morphological changes of brain tissues in rats with ischemic brain injury. Neurons appearances were basically normal in sham operation group. Cytoplasmic vacuolation and endonuclear chromatin margination were observed in model group. In Shikonin treatment group, conditions of neurons cells were improved, and degree of cell degeneration relieved than that in model group. Expression of IL-6 and IL-8 and NF-κB protein expression levels in Shikonin treatment group were lower than those in model group(P<0.01).Conclusion Shikonin has a certain effect in improvement of brain injuries in rats with ischemic brain injury.
Objective To investigate protective effect and mechanism of Shikonin on brain tissues of rats ischemic brain injury. Methods A total of 48 rats were randomly divided into sham operation group, model group and Shikonin treatment group(n=16 in each group). Rats models with ischemic brain injury were prepared to observe daily behavior, and pathological observation for brain tissues were performed. Enzyme linked immunosorbent assay was used to detect contents of interleukin-6(IL-6) and interleukin-8(IL-8), and protein expression levels of nuclear factor-κB(NF-κB) were detected by Western blot among three groups. Results In model group, animal activity was decreased, and some animals appeared restlessness; In sham operation group, neurons in cortex and hippocampal cone ordered clearly. In model group, neurons cell layers in cortex and hippocampal cone disordered with decreased density. Shikonin treatment group could improve abnormal morphological changes of brain tissues in rats with ischemic brain injury. Neurons appearances were basically normal in sham operation group. Cytoplasmic vacuolation and endonuclear chromatin margination were observed in model group. In Shikonin treatment group, conditions of neurons cells were improved, and degree of cell degeneration relieved than that in model group. Expression of IL-6 and IL-8 and NF-κB protein expression levels in Shikonin treatment group were lower than those in model group(P<0.01).Conclusion Shikonin has a certain effect in improvement of brain injuries in rats with ischemic brain injury.
引文
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[2] Prangsaengtong O,Jantaree P,Lirdprapamongkol K,et al.Shikonin Suppresses Lymphangiogenesis VIA NF-κB/HIF-1α Axis Inhibition[J].Biol Pharm Bull,2018,41(11):1659-1666.
[3] Sun M,Shinoda Y,Fukunaga K.KY-226 Protects Blood-brain Barrier Function Through the Akt/FoxO1 Signaling Pathway in Brain Ischemia[J].Neuroscience,2019,399:89-102.
[4] 涂献坤,杨卫忠,石松生,等.黄芩苷抑制缺血性脑损伤大鼠脑组织TNF-α和AQP-4表达及减轻脑损伤的研究[J].中国药理学通报,2013,29(9):1222-1225.
[5] Zhang D L,Liu X,Wang Q,et al.Downregulation of microRNA-196a attenuates ischemic brain injury in rats by directly targeting HMGA1[J].Eur Rev Med Pharmacol Sci,2019,23(2):740-748.
[6] Han X,Zheng Z,Wang C,et al.Association between MEG3/miR-181b polymorphisms and risk of ischemic stroke[J].Lipids Health Dis,2018,17(1):292.
[7] Huang J Z,Ren Y,Jiang Y,et al.GluR1 protects hypoxic ischemic brain damage via activating Akt signaling pathway in neonatal rats[J].Eur Rev Med Pharmacol Sci,2018,22(24):8857-8865.
[8] Cutler R G,Camandola S,Feldman N H,et al.Uric acid enhances longevity and endurance and protects the brain against ischemia[J].Neurobiol Aging,2019,75:159-168.
[9] Guo Z L,Li J Z,Ma Y Y,et al.Shikonin sensitizes A549 cells to TRAIL-induced apoptosis through the JNK,STAT3 and AKT pathways[J].BMC Cell Biol,2018,19(1):29.
[10] Figat R,Zgadzaj A,Geschke S,et al.Cytotoxicity and antigenotoxicity evaluation of acetylshikonin and shikonin[J].Drug Chem Toxicol,2018,21:1-8.
[11] 李哲,靳玮,李娜,等.紫草素对慢性脑低灌注损伤大鼠认知功能的保护作用[J].脑与神经疾病杂志,2018,26(5):276-279.
[12] Varma S R,Sivaprakasam T O,Arumugam I,et al.In vitro anti-inflammatory and skin protective properties of Virgin coconut oil[J].J Tradit Complement Med,2018,9(1):5-14.
[13] Xu G,Gu H,Hu B,et al.PEG-b-(PELG-g-PLL) nanoparticles as TNF-α nanocarriers:potential cerebral ischemia/reperfusion injury therapeutic applications[J].Int J Nanomedicine,2017,12:2243-2254.
[14] Rocha-Ferreira E,Kelen D,Faulkner S,et al.Systemic pro-inflammatory cytokine status following therapeutic hypothermia in a piglet hypoxia-ischemia model[J].J Neuroinflammation,2017,14(1):44.
[15] Al-Shargabi T,Govindan R B,Dave R,et al.Inflammatory cytokine response and reduced heart rate variability in newborns with hypoxic-ischemic encephalopathy [J].J Perinatol,2017,37(6):668-672.
[16] Molnar T,Pusch G,Nagy L,et al.Correlation of the L-Arginine Pathway with Thrombo-Inflammation May Contribute to the Outcome of Acute Ischemic Stroke[J].J Stroke Cerebrovasc Dis,2016,25(8):2055-2060.
[17] Gredal H,Thomsen B B,Boza-Serrano A,et al.Interleukin-6 is increased in plasma and cerebrospinal fluid of community-dwelling domestic dogs with acute ischaemic stroke[J].Neuroreport,2017,28(3):134-140.
[18] Bi Y,Zhu Y,Zhang M,et al.Effect of Shikonin on Spinal Cord Injury in Rats VIA Regulation of HMGB1/TLR4/NF-kB Signaling Pathway[J].Cell Physiol Biochem,2017,43(2):481-491.