大黄素对大鼠局灶性脑缺血的保护作用及其机制

doi:10.13481/j.1671-587x.20200323

摘要/Abstract

摘要： 目的：探讨大黄素对大鼠局灶性脑缺血的保护作用，阐明其作用机制。方法：将60只健康雄性SD大鼠随机分为假手术组（给予生理盐水），模型组（给予生理盐水，建模），阳性对照组（给予5mg·kg^-1地塞米松，建模），低、中和高剂量大黄素组（给予10、20和40 mg·kg^-1大黄素，建模）（n=10）。采用大脑中动脉阻断法（MCAO）建立局灶性脑缺血大鼠模型。检测各组大鼠行为学评分，采用TTC染色法检测各组大鼠脑梗死体积百分比，ELISA法检测各组大鼠缺血侧海马组织中肿瘤坏死因子（TNF-α）、白细胞介素1β（IL-1β）和白细胞介素6（IL-6）水平，Western blotting法检测各组大鼠缺血侧海马组织中核转录因子κB （NF-κB） p65和核转录因子κB抑制蛋白α（IκBα）蛋白表达水平。结果：假手术组大鼠未发现神经功能缺损症状和脑梗死；与假手术组比较，模型组大鼠缺血侧海马组织中IL-6、IL-1β和TNF-α水平明显升高（P<0.05），NF-κB p65蛋白表达水平明显升高（P<0.05），IκBα蛋白表达水平明显降低（P<0.05）。与模型组比较，阳性对照组，中和高剂量大黄素组大鼠行为学评分降低（P<0.05），脑梗死体积百分比明显减小（P<0.05），缺血侧海马组织中IL-6、IL-1β和TNF-α水平明显降低（P<0.05），NF-κB p65蛋白表达水平明显降低（P<0.05），IκBα蛋白表达水平明显升高（P<0.05）。与低剂量大黄素组比较，中和高剂量大黄素组大鼠行为学评分降低（P<0.05）、脑梗死体积百分比明显减小（P<0.05），缺血侧海马组织中IL-6、IL-1β和TNF-α水平明显降低（P<0.05），NF-κB p65蛋白表达水平明显降低（P<0.05），IκBα蛋白表达水平明显升高（P<0.05）。结论：大黄素对大鼠局灶性脑缺血损伤有一定的保护作用，其机制可能与抑制NF-κB信号通路和降低炎症因子分泌有关。

关键词: 大黄素, 脑缺血, 炎症因子, 核因子κB, 脑梗死

Abstract: Objective: To investigate the protective effect of emodin on the focal cerebral ischemia in the rats, and to clarify its mechanism. Methods: Sixty healthy male SD rats were randomly divided into sham operation group(given saline), model group(given saline,modeling),positive control group(give 5 mg·kg^-1 dexamethasone, modeling), low, medium and high doses of emodin groups(given 10,20,and 40 mg·kg^-1 emodin,modeling)(n=10). The middle cerebral artery occlusion(MCAO) was used to establish the rat models of focal cerebral ischemia.The behavioral scores of the rats in various groups were detected,the percentages of cerebral infarction volumes of the rats in various groups were determined by TTC staining,the levels of tumor necrosis factor (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6(IL-6) in hippocampus tissue at ischemic side of the rats in various groups were measured by enzyme linked immunosorbent assay(ELISA) method,and the expression levels of nuclear transcription factor-κB (NF-κB) p65 protein and nuclear transcription factor κB inhibitor-α(IκBα) protein in hippocampus tissue at ischemic side of the rats in various groups were detected by Western blotting method. Results: No neurological defects or cerebral infarction of the rats in sham operation group was found. Compared with sham operation group, the levels of IL-6,IL-1β, and TNF-α in hippocampus tissue of the rats in model group were significantly increased (P<0.05), while the expression level of NF-κB p65 protein was significantly increased (P<0.05), and the expression level of IκBα protein was decreased significantly (P<0.05).Compared with model group, the behavioral scores of the rats in positive control group, medium and high doses of emodin groups were significantly decreased(P<0.05),the percentages of cerebral infarction volumes were decreased(P<0.05), the levels of IL-6,IL-1β,and TNF-α in hippocampus tissue at ischemic side of the rats were significantly decreased(P<0.05), the expression levels of NF-κB p65 protein were significantly decreased(P<0.05),and the expression levels of IκBα protein were significantly increased(P<0.05).Compared with low dose of emodin group, the behavioral scores and the percentages of cerebral infarction volumes of the rats in medium and high doses of emodin groups were significantly decreased (P<0.05), the levels of IL-6,IL-1β,and TNF-α in hippocampus tissue at ischemic side of the rats were significantly decreased (P<0.05), the expression levels of NF-κB p65 protein were significantly decreased(P<0.05), and the expression levels of IκBα protein were significantly increased(P<0.05). Conclusion: Emodin has the protective effect on the focal cerebral ischemia, and its mechanism may be related to the inhibition of NF-κB signaling pathway and the secretion of inflammatory factors.

Key words: emodin, cerebral ischemia, inflammatory cytokines, nuclear transcription factor-κB, cerebral infarction

中图分类号:

R285.5

毕勇志, 马芬, 时俊霞, 宋琳琳, 李林泽. 大黄素对大鼠局灶性脑缺血的保护作用及其机制[J]. 吉林大学学报(医学版), 2020, 46(03): 569-574.

BI Yongzhi, MA Fen, SHI Junxia, SONG Linlin, LI Linze. Protective effect of emodin on focal cerebral ischemia in rats and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2020, 46(03): 569-574.

参考文献

[1] MEURER W J, BARTH B E, GADDIS G, et al. Rapid systematic review:intra-arterial thrombectomy "clot retrieval" for selected patients with acute ischemic stroke[J]. J Emerg Med, 2017, 52(2):255-261.
[2] ANTONINO T, CARLO M, ANTONIO P. Inflammation and inflammatory cell recruitment in acute cerebrovascular diseases[J]. Cur Immunol Rev, 2015, 11(1):24-32.
[3] ZHANG F, YAN C, WEI C J, et al. Vinpocetine inhibits NF-κB-dependent inflammation in acute ischemic stroke patients[J]. Transl Stroke Res, 2018, 9(2):174-184.
[4] 王刚, 王海燕, 梁宏, 等. LPS对大鼠骨髓MSCNF-κBp65及抗炎性细胞因子mRNA表达的影响[J]. 青海医学院学报, 2013, 34(3):182-185.
[5] JAHNKE G D, PRICE C J, MARR M C, et al. Developmental toxicity evaluation of emodin in rats and mice[J]. Birth Defects Res B Dev Reprod Toxicol, 2004, 71(2):89-101.
[6] 高红刚,周菊华. 大黄素抗炎作用及相关机制研究进展[J]. 济宁医学院学报, 2016, 39(5):348-352.
[7] PARK S Y, CHOI Y W, PARK G. Nrf2-mediated neuroprotection against oxygen-glucose deprivation/reperfusion injury by emodin via AMPK-dependent inhibition of GSK-3[J]. J Pharm Pharmacol, 2018, 70(4):525-535.
[8] WANG CY, ZHANG D L, MA H M, et al. Neuroprotective effects of emodin-8-O-β-d -glucoside in vivo and in vitro[J]. Eur J Pharmacol, 2007, 577(1-3):58-63.
[9] 王果, 韩书珍, 陈翔. 芹菜素对急性大鼠脑缺血再灌注后脑组织NCAM表达的影响[J]. 中华中医药学刊, 2015, 33(10):2440-2442.
[10] LONGA E Z, WEINSTEIN P R, CARLSON S, et al. Reversible middle cerebral artery occlusion without craniectomy in rats[J]. Stroke, 1989, 20(1):84-91.
[11] 杨腾腾, 石秋艳, 孙原, 等. 瑞替加滨对缺血性脑卒中模型大鼠的保护作用及机制研究[J]. 中国临床研究, 2017, 30(7):903-907.
[12] BEDERSON J B, PITTS L H, TSUJI M, et al. Rat middle cerebral artery occlusion:evaluation of the model and development of a neurologic examination[J]. Stroke, 1986, 17(3):472-476.
[13] LYDEN P, WAHLGREN N G. Mechanisms of action of neuroprotectants in stroke[J]. J Stroke Cerebrovasc Dis, 2000, 9(6Pt2):9-14.
[14] 隋念含, 蒙国懿, 杨金凤, 等. 甲基苯丙胺中毒大鼠纹状体TNF-α、IL-1β及星形胶质细胞与小胶质细胞的变化[J]. 中国药物依赖性杂志, 2018, 27(2):99-103.
[15] AMANTEA D, CERTO M, RUSSO R, et al. Early reperfusion injury is associated to MMP2 and IL-1β elevation in cortical neurons of rats subjected to middle cerebral artery occlusion[J]. Neuroscience, 2014, 277:755-763.
[16] 宋晓洁, 冯伟平, 韩雪娇,等. 血清IL-6及S-100B水平对颅脑损伤严重程度和预后评估的临床意义[J]. 现代生物医学进展, 2016, 16(20):3883-3886.
[17] LEMARCHANT S, DUNGHANA H, POMESHCHIK Y,et al. Anti-inflammatory effects of ADAMTS-4 in a mouse model of ischemic stroke[J]. Glia, 2016, 64(9):1492-1507.
[18] 王喜丰, 汪敏, 李刚,等. 中性鞘磷脂酶-2(Neutral sphingomyelin-2 N-SMase2)对大鼠脑缺血再灌注损伤中IL-6、IL-1β、TNF-α炎性因子的影响[J]. 中风与神经疾病杂志, 2018, 35(4):310-313.
[19] ORTIS F, MIANI M, COLLI M L, et al. Differential usage of NF-κB activating signals by IL-1β and TNF-α in pancreatic beta cells[J]. FEBS Lett, 2012, 586(7):984-989.
[20] 张朝弘, 刘丹彦. 雷公藤红素后处理对大鼠局灶性脑缺血再灌注损伤后脑组织NF-κB及TNF-α、IL-1β的影响[J]. 重庆医科大学学报, 2015, 40(1):37-41.
[21] 李祥,王毅,于湘友.关注神经重症患者迟发性脑缺血的治疗[J].中国实用内科杂志,2019,39(12):1040-1044.