藏药哇夏嘎对大鼠脑缺血再灌注损伤保护作用的研究
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摘要
目的：初步探讨藏药哇夏嘎(Adhatoda vasica Nees)(?)寸大鼠脑缺血再灌注损伤后的神经保护作用,并通过分析其对丙二醛(malondialdehyde, MDA)含量和基质会属蛋白酶-9(matrix metalloproteinase-9, MMP-9)表达的影响进一步探索其可能的脑保护机制。
方法：将198只健康雄性Wistar大鼠,用随机分组的方法,分为假手术组、模型组和治疗组(哇夏嘎治疗组),其中模型组和治疗组再分为脑缺血2h后再灌注6h、24h、48h、72h和120h,共11个亚组,每亚组18只大鼠。采用改良线栓法制作大鼠大脑中动脉闭塞模型,造模成功后1h即脑缺血1h后给予治疗组哇夏嘎1.5ml/100g (含生药3g)灌胃,24h后为1.5ml/100g/d,其余各组给予等量蒸馏水灌胃。在相应时相对各亚组分别进行神经功能缺损评分、红四氮唑染色测量脑梗死体积、苏木精-伊红染色(HE染色)观察组织形态学改变、原位未端标记法检测凋亡细胞数目,及大鼠血清MDA含量和脑组织MMP-9表达的变化。
结果：TTC染色显示,模型组与治疗组在右侧大脑均有梗死灶形成,且HE染色发现缺血范围内神经元胞质着色变浅,明显水肿,胞核浓缩、深染,与治疗组比较,模型组病变范围较大、程度较重。治疗组与模型组血清MDA含量和MMP-9浓度在脑缺血再灌注损伤后均有不同程度升高,并共同在48h到达峰值,且治疗组血清MDA含量、mmp-9浓度、神经功能缺损评分、脑梗死体积、凋亡细胞数目均低于同一时相模型组,与模型组相比,差异有统计学意义(P<0.05)。
结论：藏药哇夏嘎对大鼠脑缺血再灌注损伤后的神经功能缺损症状有一定的改善作用,并可减轻脑细胞水肿、脑组织变性及坏死等病理损伤,缩小脑梗死体积,抑制神经元凋亡,其作用机制可能与减轻脑缺血再灌注损伤后自由基损伤,并抑制缺血灶MMP-9的表达有关。
Objective:To observe neuroprotective effects of Adhatoda vasica Nees of Tibet medicine on cerebral ischemia reperfusion injury in rats and the influence of expression of the malondialdehyde and matrix metalloproteinase-9(mmp-9).
Method:one hundred and ninety-eight healthy male Wistar rats to establish cerebral ischemia reperfusion model with middle cerebral artery thread embolism method, which were randomly divided into the sham-operation group, model group and the treatment group(Adhatoda vasica Nees treatment group). According to reperfusion time interval of6h,24h,48h,72h and120h after cerebral ischemia reperfusion injury, The model group and the treatment group were divided into five subgroups. There were eighteen rats in each subgroups and the sham operation-group. The treatment groups were given Adhatoda vasica Nees after the cerebral ischemia in1h, at dose of1.5mL/100g/time and after24h were given1.5mL/100g/d. The other groups were given the same dose of water. And then observed neurological deficit scores, the infarct sizes, the apoptotic cells and the malondialdehyde and matrix metalloproteinase-9of every group at the different time points.
Results:The Wistar rats of sham group were healthy without any neurological deficit symptoms, and the nerve cells of the model group and treatment group have the infarct formation, and obvious edema in the zone of right cerebral. Cytoplasmic staining become shallow and nucleus stained darkly and become concentrated. The malondialdehyde and matrix metalloproteinase-9in treatment group and model group had been increased to varying degrees in the ischemia reperfusion. Compared with model group, the expression of malondialdehyde and matrix metalloproteinase-9, neurological deficit scores, infarct sizes, apoptotic cells of the treatment group decreased significantly, the difference was statistically significant (P<0.05).
Conclusion:It is hypothesized that Adhatoda vasica Nees could improve the symptoms of neurological deficit after cerebral ischemia reperfusion injury, lower degeneration and necrosis of brain tissue and other pathological damage, reduce the edema of the nerve cells, shrink the volume of cerebral infarction, inhibit the neuronal apoptosis by down-regulation the expression of malondialdehyde and matrix metalloproteinase-9. The results suggest that Adhatoda vasica Nees plays a neuroprotective role in cerebral ischemia injury in rats.
方法：将198只健康雄性Wistar大鼠,用随机分组的方法,分为假手术组、模型组和治疗组(哇夏嘎治疗组),其中模型组和治疗组再分为脑缺血2h后再灌注6h、24h、48h、72h和120h,共11个亚组,每亚组18只大鼠。采用改良线栓法制作大鼠大脑中动脉闭塞模型,造模成功后1h即脑缺血1h后给予治疗组哇夏嘎1.5ml/100g (含生药3g)灌胃,24h后为1.5ml/100g/d,其余各组给予等量蒸馏水灌胃。在相应时相对各亚组分别进行神经功能缺损评分、红四氮唑染色测量脑梗死体积、苏木精-伊红染色(HE染色)观察组织形态学改变、原位未端标记法检测凋亡细胞数目,及大鼠血清MDA含量和脑组织MMP-9表达的变化。
结果：TTC染色显示,模型组与治疗组在右侧大脑均有梗死灶形成,且HE染色发现缺血范围内神经元胞质着色变浅,明显水肿,胞核浓缩、深染,与治疗组比较,模型组病变范围较大、程度较重。治疗组与模型组血清MDA含量和MMP-9浓度在脑缺血再灌注损伤后均有不同程度升高,并共同在48h到达峰值,且治疗组血清MDA含量、mmp-9浓度、神经功能缺损评分、脑梗死体积、凋亡细胞数目均低于同一时相模型组,与模型组相比,差异有统计学意义(P<0.05)。
结论：藏药哇夏嘎对大鼠脑缺血再灌注损伤后的神经功能缺损症状有一定的改善作用,并可减轻脑细胞水肿、脑组织变性及坏死等病理损伤,缩小脑梗死体积,抑制神经元凋亡,其作用机制可能与减轻脑缺血再灌注损伤后自由基损伤,并抑制缺血灶MMP-9的表达有关。
Objective:To observe neuroprotective effects of Adhatoda vasica Nees of Tibet medicine on cerebral ischemia reperfusion injury in rats and the influence of expression of the malondialdehyde and matrix metalloproteinase-9(mmp-9).
Method:one hundred and ninety-eight healthy male Wistar rats to establish cerebral ischemia reperfusion model with middle cerebral artery thread embolism method, which were randomly divided into the sham-operation group, model group and the treatment group(Adhatoda vasica Nees treatment group). According to reperfusion time interval of6h,24h,48h,72h and120h after cerebral ischemia reperfusion injury, The model group and the treatment group were divided into five subgroups. There were eighteen rats in each subgroups and the sham operation-group. The treatment groups were given Adhatoda vasica Nees after the cerebral ischemia in1h, at dose of1.5mL/100g/time and after24h were given1.5mL/100g/d. The other groups were given the same dose of water. And then observed neurological deficit scores, the infarct sizes, the apoptotic cells and the malondialdehyde and matrix metalloproteinase-9of every group at the different time points.
Results:The Wistar rats of sham group were healthy without any neurological deficit symptoms, and the nerve cells of the model group and treatment group have the infarct formation, and obvious edema in the zone of right cerebral. Cytoplasmic staining become shallow and nucleus stained darkly and become concentrated. The malondialdehyde and matrix metalloproteinase-9in treatment group and model group had been increased to varying degrees in the ischemia reperfusion. Compared with model group, the expression of malondialdehyde and matrix metalloproteinase-9, neurological deficit scores, infarct sizes, apoptotic cells of the treatment group decreased significantly, the difference was statistically significant (P<0.05).
Conclusion:It is hypothesized that Adhatoda vasica Nees could improve the symptoms of neurological deficit after cerebral ischemia reperfusion injury, lower degeneration and necrosis of brain tissue and other pathological damage, reduce the edema of the nerve cells, shrink the volume of cerebral infarction, inhibit the neuronal apoptosis by down-regulation the expression of malondialdehyde and matrix metalloproteinase-9. The results suggest that Adhatoda vasica Nees plays a neuroprotective role in cerebral ischemia injury in rats.
引文
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