益脉康片对放射性脑损伤小鼠保护作用及机制研究
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摘要
目的探讨益脉康片对放射性脑损伤小鼠的保护作用及主要作用机制。方法将50只BALB/c小鼠随机分为对照组、模型组和益脉康片低、中、高剂量(4.30、8.61、17.22 mg/kg)组,采用伽玛刀对除对照组外其余各组小鼠进行造模,连续7 d。造模结束后,益脉康片组ig给药,对照组及模型组给予等量生理盐水,连续给药2周,1次/d。末次给药24 h后,采用Morris法检测各组小鼠的学习及记忆能力,并对各组小鼠海马体CA1区进行TUNEL染色及尼氏染色,采用免疫组化法检测各组小鼠兔抗烟酰胺腺嘌呤二核苷酸磷酸酶氧化酶(NOX4)的表达情况,并对其进行半定量分析。结果第2～5天,对照组和益脉康片低、中、高剂量组潜伏期均明显低于模型组(P<0.05);与模型组比较,对照组和益脉康片低、中、高剂量组第Ⅲ象限时间及穿越平台次数均明显增加(P<0.05);尼氏染色中,与模型组比较,益脉康片低、中、高剂量小鼠海马体中神经细胞核核仁不清晰及细胞形态异常程度均明显减轻;TUNEL染色中,与模型组比较,益脉康片低、中、高剂量组均仅有呈散在的少量阳性染色神经元;免疫组化中,与模型组比较,益脉康片低、中、高剂量组中NOX4阳性染色均明显弱于模型组,且相对表达量均明显低于模型组(P<0.05)。结论益脉康片对放射性脑损伤小鼠具有较好的保护作用,其作用机制可能与有效抑制小鼠海马体中NOX4表达有关。
Objective To study the protective effects and main mechanism of Yimaikang tablet on radiation-induced brain injury in mice. Methods A total of 50 BALB/c mice were divided into control group, model group, and Yimaikang tablet low, middle and high dose (4.30, 8.61, and 17.22 mg/kg) treatment groups. The gamma knife was used to establish the radiation-induced brain injury model for continuous 7 d. After the modeling, mice in Yimaikang tablet group were ig administrated corresponding drugs, the other groups were treated by equivalence saline, lasted 2 weeks, once per day. 24 hours after the last gavage, the Morris method was used to test the learning and memory ability of mice in each group. The TUNEL staining and Nissl's staining were performed on hippocampus CA1 region of mice. The immunohistochemical method was performed to observe the expression of NOX4 in mice,and the semi-quantitative analysis was also carried out. Results In the training of 2—5 d, the incubation periods of control group and low, middle, and high dose groups of Yimaikang tablet were significantly lower than model group (P < 0.05); Compared with the model group, the third quadrant residence time and times of crossing the platform in control group and Yimaikang tablet groups were significantly increased (P < 0.05). The TUNEL staining showed that there were small numbers of positive staining neurons in Yimaikang tablet groups when compared with the model group. The Nissl's staining showed that, compared with the model group,the cell morphological abnormalities and nucleoli blurring of neurocytes in the hippocampus of mice were significantly alleviated in low, middle, and high dose groups of Yimaikang tablet. From the results of immunohistochemical analysis, we found that the NOX4 positive stainings in Yimaikang tablet groups were significantly less than the model group, and the relative expression were significantly lower than model group (P < 0.05). Conclusion Yimaikang tablet has a good protective effects on radiation-induced brain injury in mice and the mechanism may be related to the effective inhibition of NOX4 expression in the hippocampus of mice.
Objective To study the protective effects and main mechanism of Yimaikang tablet on radiation-induced brain injury in mice. Methods A total of 50 BALB/c mice were divided into control group, model group, and Yimaikang tablet low, middle and high dose (4.30, 8.61, and 17.22 mg/kg) treatment groups. The gamma knife was used to establish the radiation-induced brain injury model for continuous 7 d. After the modeling, mice in Yimaikang tablet group were ig administrated corresponding drugs, the other groups were treated by equivalence saline, lasted 2 weeks, once per day. 24 hours after the last gavage, the Morris method was used to test the learning and memory ability of mice in each group. The TUNEL staining and Nissl's staining were performed on hippocampus CA1 region of mice. The immunohistochemical method was performed to observe the expression of NOX4 in mice,and the semi-quantitative analysis was also carried out. Results In the training of 2—5 d, the incubation periods of control group and low, middle, and high dose groups of Yimaikang tablet were significantly lower than model group (P < 0.05); Compared with the model group, the third quadrant residence time and times of crossing the platform in control group and Yimaikang tablet groups were significantly increased (P < 0.05). The TUNEL staining showed that there were small numbers of positive staining neurons in Yimaikang tablet groups when compared with the model group. The Nissl's staining showed that, compared with the model group,the cell morphological abnormalities and nucleoli blurring of neurocytes in the hippocampus of mice were significantly alleviated in low, middle, and high dose groups of Yimaikang tablet. From the results of immunohistochemical analysis, we found that the NOX4 positive stainings in Yimaikang tablet groups were significantly less than the model group, and the relative expression were significantly lower than model group (P < 0.05). Conclusion Yimaikang tablet has a good protective effects on radiation-induced brain injury in mice and the mechanism may be related to the effective inhibition of NOX4 expression in the hippocampus of mice.
引文
[1]杨宇,黄艳芳,周智慧,等.放射性脑损伤的MRI及灌注成像分析[J].中国CT和MRI杂志,2016,14(6):1-2,12.
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[4]陈泊霖,孙熠,梁宾,等.大鼠放射性脑损伤所致血脑屏障通透性改变与EBA及VEGF表达的相关性研究[J].天津医药,2016,44(6):691-693,651.
[5]孙有利,辛庆锋,李超彦,等.丹参酮ⅡA对放射性脑损伤小鼠的神经保护作用及机制研究[J].中药药理与临床,2017,33(1):66-70.
[6]赵勇,季敏,秦文燕.益脉康分散片联合胰激肽原酶治疗糖尿病视网膜病变的疗效观察[J].现代药物与临床,2016,33(9):1434-1438.
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[11]李俊晨,李国华,田野,等.放射性脑损伤的MRI研究进展[J].中华放射肿瘤学杂志,2017,26(1):98-102.
[12]周东晓,郭俊杰,谢颖,等.放射性脑损伤的血管损伤机制及贝伐珠单抗临床应用的研究进展[J].国际神经病学神经外科学杂志,2017,44(2):226-229.
[13]Panickar K S,Anderson R A.Effect of polyphenols on oxidative stress and mitochondrial dysfunction in neuronal death and brain edema in cerebral ischemia[J].Int J Mol Sci,2011,12(11):8181-8207.
[14]Carbone F,Teixeira P C,Braunersreuther V,et al.Pathophysiology and treatments of oxidative injury in ischemic stroke:focus on the phagocytic NADPHoxidase 2[J].Antioxid Redox Signal,2015,23(5):460-489.
[15]Nishimura A,Ago T,Kuroda J,et al.Detrimental role of pericyte Nox4 in the acute phase of brain ischemia[J].JCereb Blood Flow Metab,2016,36(6):1143-1154.
[16]雷海云,黄崇柯.银杏达莫联合益脉康治疗视网膜静脉阻塞的临床效果[J].国际眼科杂志,2009,9(10):1989-1990.
[17]张綦慧,张丽,牛焕敏,等.灯盏细辛制剂在脑血管病治疗中的应用[J].吉林中医药,2016,36(9):941-943,944.
[18]夏靓.灯盏细辛的化学成分及其制剂的研究进展[J].中国药房,2016,27(1):111-113.
[19]Panagiotakos G,Alshamy G,Chan B,et al.Long-term impact of radiation on the stem cell and oligodendrocyte precursors in the brain[J].PLoS One,2007,2(7):e588.
[2]熊耀祖,孙阳,涂彧,等.大鼠放射性脑损伤模型中Cox-2 mRNA及蛋白的动态变化研究[J].中华放射医学与防护杂志,2016,36(1):24-27.
[3]李洲,马林,陈旺生,等.鼻咽癌放射性脑损伤的MR动态磁敏感对比灌注成像[J].中国医学影像学杂志,2016,24(8):561-564.
[4]陈泊霖,孙熠,梁宾,等.大鼠放射性脑损伤所致血脑屏障通透性改变与EBA及VEGF表达的相关性研究[J].天津医药,2016,44(6):691-693,651.
[5]孙有利,辛庆锋,李超彦,等.丹参酮ⅡA对放射性脑损伤小鼠的神经保护作用及机制研究[J].中药药理与临床,2017,33(1):66-70.
[6]赵勇,季敏,秦文燕.益脉康分散片联合胰激肽原酶治疗糖尿病视网膜病变的疗效观察[J].现代药物与临床,2016,33(9):1434-1438.
[7]孙敬方.动物实验方法学[M].北京:人民卫生出版社,2001.
[8]Tang Y M,Rong X M,Hu W H,et al.Effect of edaravone on radiation-induced brain necrosis in patients with nasopharyngeal carcinoma after radiotherapy:a randomized controlled trial[J].J Neurooncol,2014,120(2):441-447.
[9]武海霞,吴志刚,刘红彬,等.Morris水迷宫实验在空间学习记忆研究中的应用[J].神经药理学报,2014,4(5):30-35.
[10]胡兰花,于韬,徐婷婷,等.动态磁敏感对比增强MRI和动态对比增强MRI鉴别诊断胶质瘤复发和放射性脑损伤[J].中国医学影像技术,2017,33(1):11-16.
[11]李俊晨,李国华,田野,等.放射性脑损伤的MRI研究进展[J].中华放射肿瘤学杂志,2017,26(1):98-102.
[12]周东晓,郭俊杰,谢颖,等.放射性脑损伤的血管损伤机制及贝伐珠单抗临床应用的研究进展[J].国际神经病学神经外科学杂志,2017,44(2):226-229.
[13]Panickar K S,Anderson R A.Effect of polyphenols on oxidative stress and mitochondrial dysfunction in neuronal death and brain edema in cerebral ischemia[J].Int J Mol Sci,2011,12(11):8181-8207.
[14]Carbone F,Teixeira P C,Braunersreuther V,et al.Pathophysiology and treatments of oxidative injury in ischemic stroke:focus on the phagocytic NADPHoxidase 2[J].Antioxid Redox Signal,2015,23(5):460-489.
[15]Nishimura A,Ago T,Kuroda J,et al.Detrimental role of pericyte Nox4 in the acute phase of brain ischemia[J].JCereb Blood Flow Metab,2016,36(6):1143-1154.
[16]雷海云,黄崇柯.银杏达莫联合益脉康治疗视网膜静脉阻塞的临床效果[J].国际眼科杂志,2009,9(10):1989-1990.
[17]张綦慧,张丽,牛焕敏,等.灯盏细辛制剂在脑血管病治疗中的应用[J].吉林中医药,2016,36(9):941-943,944.
[18]夏靓.灯盏细辛的化学成分及其制剂的研究进展[J].中国药房,2016,27(1):111-113.
[19]Panagiotakos G,Alshamy G,Chan B,et al.Long-term impact of radiation on the stem cell and oligodendrocyte precursors in the brain[J].PLoS One,2007,2(7):e588.