基于氧化应激和炎症介质反应探讨草木樨提取物对急性脑缺血组织保护作用的机制研究  

Protective effect and mechanism of Melilotus officinalis extracts on acute cerebral ischemia based on oxidative stress and inflammatory mediator responses

作　　者：林晓伟 邓明尧 孙冕 林雅明 Lin Xiaowei;Deng Mingyao;Sun Mian(Department of Emergency,Sanya People's Hospital,Sanya Hainan 572000)

机构地区：[1]海南省三亚市人民医院急诊科,572000

出　　处：《卒中与神经疾病》2022年第4期317-323,共7页Stroke and Nervous Diseases

基　　金：2018年海南省卫生计生行业科研项目(1802330169A2001)。

摘　　要：目的评估草木樨提取物对急性脑缺血组织的保护作用及保护机制。方法将50只雄性Wistar大鼠随机分为假手术组、模型组、草木樨低、中、高剂量组,每组各10只;除假手术组外,其余各组大鼠通过改良Longa法建立急性脑缺血模型;对草木樨低、中、高剂量组脑缺血大鼠分别给予100、250、500 mg/kg草木樨治疗,假手术组及模型组给予等量生理盐水;采用2,3,5氯化三苯基四氮唑染色法(Triphenyltetrazolium chloride,TTC)检测大鼠脑梗死体积;采用Longa评分法评估神经功能损伤评分;采用放射免疫测定法检测血浆6-酮-前列素F1a(6-keto-prostaglandin F1a,6-keto-PGF1a)及血栓素B2(Thromboxane B2,TXB2)水平;采用酶联免疫吸附实验(Enzyme linked immunosorbent assay,ELISA)试剂盒检测肿瘤坏死因子a(Tumor necrosis factor-alpha,TNF-a)、白细胞介素10(Interleukin-10,IL-10)水平;采用分光光度计评估超氧化物歧化酶(Superoxidedismutase,SOD)及谷胱甘肽过氧化物酶(Glutatheione peroxidase,GSH-Px)活性;采用原位末端凋亡法(TdT-mediated dUTP nick-end labeling,TUNEL)检测脑组织神经元凋亡情况;采用蛋白质印迹法(Western blot)检测B淋巴细胞瘤-2基因(B-cell lymphoma-2,Bcl-2)及Bcl-2-相关X(Bcl-2-Associated X,Bax)蛋白相对表达水平。结果与模型组比较,草木樨低、中、高剂量组的脑梗死体积和神经功能缺损评分显著降低(P<0.05),且随着草木樨给药剂量的增加,各组大鼠脑梗死体积及神经功能缺损评分逐渐降低(P<0.05)。与模型组比较,草木樨低、中、高剂量组血浆6-keto-PGF1a水平显著增高(P<0.05),血浆TXB2水平显著降低(P<0.05),且随着草木樨给药剂量的增加,各组大鼠血浆6-keto-PGF1a水平逐渐增高(P<0.05)、血浆TXB2水平逐渐降低(P<0.05)。与模型组比较,草木樨低、中、高剂量组TNF-a水平显著降低(P<0.05),IL-10,SOD,GSH-PX水平显著升高(P<0.05),且随着草木樨给药剂量的增加,各组大鼠TNF-a水平逐渐降�Objective To evaluate the protective effect and mechanism of Melilotus officinalis extracts on acute cerebral ischemia based on oxidative stress and inflammatory mediator responses.Methods Fifty male Wistar rats were randomly divided into sham group,model group,and low-dose,medium-dose,high-dose Melilotus officinalis groups,with 10 rats in each group.Acute cerebral ischemia models were established by modified Longa method.Rats in the low-dose,medium-dose,high-dose groups were treated with 100 mg/kg,250 mg/kg,or 500 mg/kg Melilotus officinalis extracts respectively.Rats in sham operation group and model group were treated with the same dose of normal saline.The cerebral infarction volume was measured by TTC method and neurological deficit score was estimated by Longa scoring method.6-Keto-PGF1αand TXB2 concentrations in plasma were estimated by radioimmunoassay.TNF-αand IL-10 concentrations in plasma were estimated by ELISA.The activity of SOD and GSH-Px were estimated by spectrophotometer.The apoptosis of ischemic neurons in cerebral infarction tissues was detected by TUNEL.The expression of Bcl-2 and Bax was detected by Western blot.Results Compared with the model group,the infarct volume,neurological deficit score,TXB2 concentration in plasma,the level of TNF-α,apoptosis rate and Bax expression were significantly decreased in low-dose,medium-dose,high-dose Melilotus officinalis treatment groups(P<0.05)in a dose-dependent manner.However,6-Keto-PGF1αconcentration in plasma,the levels of IL-10,SOD,GSH-PXand Bcl-2 were significantly increased(P<0.05)in Melilotus officinalis groups in a dose-dependent way.Conclusion Melilotus officinalis ameliorates apoptosis of brain tissues in cerebral ischemic rats by decreasing cerebral thrombosis,oxidative stress,and inflammatory mediator responses.Melilotus officinalis has a protective effect on ischemic brain tissue in a dose-dependent manner.

关 键 词：草木樨 脑缺血 氧化应激 细胞因子 细胞凋亡 

分 类 号：R743.3[医药卫生—神经病学与精神病学]