滋水清肝饮加味对MCAO大鼠脑保护机制的研究
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摘要
目的研究滋水清肝饮加味对急性脑缺血再灌注损伤受损脑组织的保护机制。方法 50只大鼠随机分为5组:中药高剂量组、中药低剂量组、西药组、模型组、正常组,每组各10只。采用线栓法制备局灶性脑缺血模型大鼠,中药高、低剂量组分别按成人每千克体质量剂量的12倍、6倍给予灌胃,西药组给予脑复康,按照50 mg/100 (g·d)计算,模型组和正常组给予同等量生理盐水,每日1次,共7 d。在造模成功以后24 h给药,水和饲料自由摄取。分别对50只大鼠于治疗前、治疗后进行神经功能缺损评分、平衡木行走实验评分。在给药周期结束后24 h,对大鼠脑组织完成切片、染色、拍照,进行脑梗死体积比和缺血侧大脑水肿度测定。此外选取包含缺血区皮层部位的脑切片,经脱水、包埋、切片,用于免疫组化检测肿瘤坏死因子-α(TNF-α)。结果治疗后神经功能障碍评分、平衡木行走实验评分中药高剂量组、西药组和模型组比较明显减低(P <0.05),中药低剂量组最为显著(P <0.01);中药高剂量组、中药低剂量组、西药组与模型组比较脑梗死体积明显降低(P <0.01);中药低剂量组、西药组大脑缺血侧水肿程度最为显著(P <0.01)。模型组与正常组比较,TNF-α表达显著升高;中药高、低剂量组、西药组与模型组比较,TNF-α表达均明显下降。结论滋水清肝饮加味能降低MCAO大鼠神经功能障碍评分、平衡木行走实验得分,改善MCAO大鼠神经功能障碍;减轻MCAO大鼠脑梗死的体积比和水肿程度。滋水清肝饮加味能够起到脑缺血再灌注后神经保护的作用。
Objective: To study the protective mechanism of Zishui Qinggan Decoction on damaged brain tissue after acute cerebral ischemia reperfusion injury. Methods: 50 rats were randomly divided into five groups: highdose group,low-dose group,Western medicine group,the model group and the normal group,10 rats in each. The high-dose and low-dose groups were given intragastric administration 12 times and 6 times the dose of adult kg body weight,and Western medicine group was given brain rehabilitation,50 mg/100 g per day. The model and normal group were given the same amount of physiological saline,once a day for 7 days. After successful molding,24 h administration,solid feed and water were free taken. Neurological dysfunction scores and balance beam walking test scores were performed on 50 rats before and after treatment. After the end of 24 h administration cycle,the brain tissue sections of the rats were taken,stained and photographed to determine the cerebral infarction volumetric ratio and ischemic side brain edema. In addition,brain sections containing the ischemic cortex were selected,dehydrated,embedded and sectioned for immunohistochemistry to detect TNF-α. Results: After treatment,the scores of neurological impairment and balance wood walking test in high dose group,Western medicine group and the model group were significantly reduced(P < 0.05),and those in low dose group were the most significant(P < 0.01). Compared with the model group,the volume of cerebral infarction in the high dose group,the low dose group and Western medicine group decreased significantly(P < 0.01). The degree of cerebral ischemic edema was the most significant in the low dose group and Western medicine group(P < 0.01). Compared with the normal group,the expression of TNF-α in the model group was significantly higher,while that in the high and low dose groups and Western medicine group was significantly lower than that in the model group. Conclusion:Zishui Qinggan Decoction can reduce the neurological dysfunction scores,balance wood walking experimental scores,improve the neurological dysfunction,reduce the the volume ratio and edema degree of cerebral infarction in MCAO rats. Zishui Qinggan Decoction can play a neuroprotective role after cerebral ischemia reperfusion.
Objective: To study the protective mechanism of Zishui Qinggan Decoction on damaged brain tissue after acute cerebral ischemia reperfusion injury. Methods: 50 rats were randomly divided into five groups: highdose group,low-dose group,Western medicine group,the model group and the normal group,10 rats in each. The high-dose and low-dose groups were given intragastric administration 12 times and 6 times the dose of adult kg body weight,and Western medicine group was given brain rehabilitation,50 mg/100 g per day. The model and normal group were given the same amount of physiological saline,once a day for 7 days. After successful molding,24 h administration,solid feed and water were free taken. Neurological dysfunction scores and balance beam walking test scores were performed on 50 rats before and after treatment. After the end of 24 h administration cycle,the brain tissue sections of the rats were taken,stained and photographed to determine the cerebral infarction volumetric ratio and ischemic side brain edema. In addition,brain sections containing the ischemic cortex were selected,dehydrated,embedded and sectioned for immunohistochemistry to detect TNF-α. Results: After treatment,the scores of neurological impairment and balance wood walking test in high dose group,Western medicine group and the model group were significantly reduced(P < 0.05),and those in low dose group were the most significant(P < 0.01). Compared with the model group,the volume of cerebral infarction in the high dose group,the low dose group and Western medicine group decreased significantly(P < 0.01). The degree of cerebral ischemic edema was the most significant in the low dose group and Western medicine group(P < 0.01). Compared with the normal group,the expression of TNF-α in the model group was significantly higher,while that in the high and low dose groups and Western medicine group was significantly lower than that in the model group. Conclusion:Zishui Qinggan Decoction can reduce the neurological dysfunction scores,balance wood walking experimental scores,improve the neurological dysfunction,reduce the the volume ratio and edema degree of cerebral infarction in MCAO rats. Zishui Qinggan Decoction can play a neuroprotective role after cerebral ischemia reperfusion.
引文
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[12]宋蒙蒙,田立,孟庆阳,等.自拟滋水清肝饮加减治疗短暂性后循环脑缺血发作(阴虚阳亢证)的临床观察[J].中国中医急症,2017,26(4):725-727.
[13]刘晓婷,田立.滋水清肝饮临床应用体会[J].中医药导报,2016,22(9):108-109.
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[15]傅宝君.滋水清肝饮临床应用举隅[J].浙江中医杂志,2009,44(8):586-587.
[2]Altumbabic M,Peeling J,Bigio MR,et al.Intracerebral hemorrhage in the rat:effects of hematoma aspiration[J].Stroke,1998,29(7):1917-1923.
[3]Feeney DM,Gonzalez A,Law WA.Amphetamine,haloperidol and experience interact to affect the rate of recovery after motor cortex injury[J].Science,1982,217:855-857.
[4]Hua Q,Zhu XL,Li PT,et al.The inhibitory effects of cholalicacid and hyodeoxycholalic acid on the expression of TNFalpha and IL-1beta after cerebral ischemia in rats[J].Arch Pharm Res,2009,32(1):65-73.
[5]张勇,刘玲,王海燕,等.涤痰汤对大鼠局灶性脑缺血再灌注损伤的神经保护作用[J].湖北中医杂志,2005,27(3):6-8.
[6]Buja LM.Myocardial ischemia and reperlusion injury[J].Cardiovasc Pathol,2005(14):170-175.
[7]Martinmez Vila E,Sieira PI.Acute stroke therapy:translating preclinical neuroprotection to therapeutic reality[J].Cerebrovasc Dis,2001,11(5):60-70.
[8]王小琴,邹玉安,郭春燕.脑缺血预处理和脑缺血耐受机制的研究进展[J].神经药理学报,2015,5(2):30-37.
[9]王亚茹,胡建鹏,王建,等.中风病病因病机理论的形成与发展[J].中医药临床杂志,2017,4(14):303-306.
[10]俞腾云,吴艳华,孙寒静,等.缺血性脑卒中中医的病因病机关系[J].吉林中医药,2016,5(27):328-331.
[11]胡龙涛,蔡芳妮,王亚丽.中风病病因病机探析[J].中西医结合心脑血管病杂志,2017,4(10):883-885.
[12]宋蒙蒙,田立,孟庆阳,等.自拟滋水清肝饮加减治疗短暂性后循环脑缺血发作(阴虚阳亢证)的临床观察[J].中国中医急症,2017,26(4):725-727.
[13]刘晓婷,田立.滋水清肝饮临床应用体会[J].中医药导报,2016,22(9):108-109.
[14]卢宏福.滋水清肝饮的临床应用[J].中医临床研究,2015,17(17):103-104.
[15]傅宝君.滋水清肝饮临床应用举隅[J].浙江中医杂志,2009,44(8):586-587.