脑血疏口服液灌胃对帕金森病模型大鼠黑质内神经炎症反应的影响
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摘要
目的:研究脑血疏口服液对帕金森病(PD)模型大鼠黑质内神经炎症的抑制作用,以及对黑质多巴胺(DA)能神经元的保护作用。方法:应用立体定向注射技术,向大鼠脑右侧纹状体内注射6-羟多巴胺制备PD模型。将PD模型大鼠随机分为PD组、雷沙吉兰组和脑血疏口服液高、中、低剂量干预组,每组10只;另选10只正常大鼠作为对照组。采用免疫组织化学法检测各组大鼠黑质DA能神经元(酪氨酸羟化酶(TH)阳性)、小胶质细胞(CD11b阳性)、星形胶质细胞(GFAP阳性)形态变化。Western Blot法检测中脑黑质环氧化酶-2(COX-2)和肿瘤坏死因子-α(TNF-α)的变化。结果:大鼠行为学检测和黑质TH免疫组织化学染色显示,中剂量组的干预效果最好。免疫组织化学显示,与对照组相比,PD组的星形胶质细胞、小胶质细胞、TNF-α阳性细胞和COX-2阳性细胞均明显增加,TH阳性细胞数量明显减少,经中剂量的脑血疏口服液干预后,两种胶质细胞、TNF-α和COX-2阳性细胞数量均减少,TH阳性细胞增多。Western Blot显示,与PD组比较,中剂量组和雷沙吉兰组GFAP、CD11b、TNF-α和COX-2蛋白表达量显著减少(P<0.05),而中剂量组与雷沙吉兰组比较上述蛋白表达均无显著差异(P>0.05)。结论:脑血疏口服液可能通过抑制PD大鼠黑质胶质细胞的异常激活,减少炎性因子分泌而发挥神经保护作用。
Objective:To investigate the anti-inflammatory and neuroprotective effects of Naoxueshu Oral Liquid on6-OHDA rat model of Parkinson's disease(PD).Methods:Male adult Sprague-Dawley rats were stereotactically injected with 6-hydroxydopamine(6-OHDA) into the right striatum.PD rats were randomly divided into PD group,Rasagiline group,low-,medium-and high-dose group.Ten rats were in each group.Ten adult normal rats were injected with saline instead of 6-OHDA as the control group.The morphological changes of dopaminergic neurons(tyrosine hydroxylase positive,TH-positive),astrocytes(GFAP-positive) and microglia(CD11b-positive) in substantianigra of rats in each group were examined by immunohistochemical methods.The expression changes of tumor necrosis factor-α(TNF-α) and cyclooxygenase-2(COX-2) were detected by Western Blot.Results:The results of behavioral testing and TH-immunoac-tive cells examination showed that the medium-dose Naoxueshu oral Liquid had the best therapeutic effects on PD rats(P < 0.05).Compared with the control group,the number of GFAP-,CD11b-,TNF-α-,COX-2-positive cells in substantianigra of the model group increased significantly,and the numbers of TH-positive cells was significantly decreased in PD model group.The numbers of astrocyte,microglia,TNF-α-and COX-2-positive cells were reduced,and the number of TH-positive cell increased in medium-dose group.The Western Blot showed that the expressions of GFAP,CD11 b,TNF-α and COX-2 in medium-dose group and Rasagiline group were significantly decreased compared with the PD group(P < 0.05).There were no differences between the medium-dose group and Rasagiline group(P > 0.05).Conclusion:Naoxueshu oral liquid might play a neuroprotective role by inhibiting abmormol activation of microglia and astrocytes,alleviating the inflammatory response in substantia nigra of PD rats.
Objective:To investigate the anti-inflammatory and neuroprotective effects of Naoxueshu Oral Liquid on6-OHDA rat model of Parkinson's disease(PD).Methods:Male adult Sprague-Dawley rats were stereotactically injected with 6-hydroxydopamine(6-OHDA) into the right striatum.PD rats were randomly divided into PD group,Rasagiline group,low-,medium-and high-dose group.Ten rats were in each group.Ten adult normal rats were injected with saline instead of 6-OHDA as the control group.The morphological changes of dopaminergic neurons(tyrosine hydroxylase positive,TH-positive),astrocytes(GFAP-positive) and microglia(CD11b-positive) in substantianigra of rats in each group were examined by immunohistochemical methods.The expression changes of tumor necrosis factor-α(TNF-α) and cyclooxygenase-2(COX-2) were detected by Western Blot.Results:The results of behavioral testing and TH-immunoac-tive cells examination showed that the medium-dose Naoxueshu oral Liquid had the best therapeutic effects on PD rats(P < 0.05).Compared with the control group,the number of GFAP-,CD11b-,TNF-α-,COX-2-positive cells in substantianigra of the model group increased significantly,and the numbers of TH-positive cells was significantly decreased in PD model group.The numbers of astrocyte,microglia,TNF-α-and COX-2-positive cells were reduced,and the number of TH-positive cell increased in medium-dose group.The Western Blot showed that the expressions of GFAP,CD11 b,TNF-α and COX-2 in medium-dose group and Rasagiline group were significantly decreased compared with the PD group(P < 0.05).There were no differences between the medium-dose group and Rasagiline group(P > 0.05).Conclusion:Naoxueshu oral liquid might play a neuroprotective role by inhibiting abmormol activation of microglia and astrocytes,alleviating the inflammatory response in substantia nigra of PD rats.
引文
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[17]王晶,蒋璐,谭展鹏,等.黄芪注射液联合血必净注射液对缺血性脑卒中急性期神经保护作用的研究[J].中西医结合心脑血管病杂志,2016,14:349-351.
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[21]Pandurangan K,Krishnappan V,Subramanian V,et al.Anti-inflammatory effect of certain dimethoxy flavones[J].Inflammopharmacology,2015,23:307-317.
[2]Su X,Federoff HJ.Immune responses in Parkinson’s disease:interplay between central and peripheral immune systems[J].Biomed Res Int,2014,2014:275178.
[3]Pal R,Tiwari PC,Nath R,Pant KK.Role of neuroinflammation and latent transcription factors in pathogenesis of Parkinson’s disease[J].Neurol Res,2016,38:1111-1122.
[4]艾鑫,刘翠.脑血疏口服液对脑出血大鼠血肿体积的影响及神经功能的保护作用[J].中西医结合心脑血管病杂志,2014,12:859-861.
[5]张选国,王凌.脑血疏口服液对气虚血瘀型急性脑梗死疗效及炎性指标的影响[J].中西医结合心脑血管病杂志,2016,14:299-300.
[6]李晓健,庄文欣,吕娥,等.香椿子多酚对6-OHDA所致帕金森病大鼠模型的抗炎和神经保护作用[J].神经解剖学杂志,2016,32:653-659.
[7]Ouchi Y,Yagi S,Yokokura M,et al.Neuroinflammation in the living brain of Parkinson’s disease[J].Parkinsonism Relat Disord,2009,15:S200-S204.
[8]De Virgilio A,Greco A,Fabbrini G,et al.Parkinson’s disease:autoimmunity and neuroinflammation[J].Autoimmun Rev,2016,15:1005-1011.
[9]Hirsch EC,Vyas S,Hunot S.Neuroinflammation in Parkinson’s disease[J].Parkinsonism Relat Disord,2011,18:S210-S212.
[10]Niranjan R.The role of inflammatory and oxidative stress mechanisms in the pathogenesis of Parkinson’s disease:Focus on astrocytes[J].Mol Neurobiol,2013,49:28-38.
[11]庄文欣,付文玉,吕娥,等.帕金森病大鼠黑质小胶质细胞及星形胶质细胞的变化[J].解剖学报,2010,41:344-348.
[12]Ransohoff RM,Perry VH.Microglial physiology:unique stimuli,specialized responses[J].Annu Rev Immunol,2009,27:119-145.
[13]Iravani MM,Sadeghian M,Leung CC,et al.Lipopolysaccharideinduced nigral inflammation leads to increased IL-1βtissue content and expression of astrocytic glial cell line derived neurotrophic factor[J].Neurosci Lett,2012,510:138-142.
[14]郭晓庆,孙佳明,张辉.水蛭的化学成分与药理作用[J].吉林中医药,2015,35:47-50.
[15]Karabiyikoglu M,Hua Y,Keep RF,et al.Intracerebral hirudin injection attenuates ischemic damage and neurologic deficits without altering local cerebral blood flow[J].J Cereb Blood Flow Metab,2004,24:159-166.
[16]谢靖,高惠春,郑玺.黄芪甲苷对急性脑出血大鼠神经功能的影响[J].中国病理生理杂志,2016,32:1905-1908.
[17]王晶,蒋璐,谭展鹏,等.黄芪注射液联合血必净注射液对缺血性脑卒中急性期神经保护作用的研究[J].中西医结合心脑血管病杂志,2016,14:349-351.
[18]Liu HS,Shi HL,Huang F,et al.Astragaloside IV inhibits microglia activation via glucocorticoid receptor mediated signaling pathway[J].Sci Rep,2016,6:19137.
[19]Yue R,Li X,Chen B,et al.Astragaloside IV attenuates glutamateinduced neurotoxicity in PC12 cells through Raf-MEK-ERK pathway[J].PLo S One,2015,10:e0126603.
[20]HanákováZ,Ho2ek J,Kutil Z,et al.Anti-inflammatory activity of natural geranylated flavonoids:cyclooxygenase and lipoxygenase inhibitory properties and proteomic analysis[J].J Nat Prod,2017Mar 21.doi:10.1021/acs.jnatprod.6b01011.[Epub ahead of print]
[21]Pandurangan K,Krishnappan V,Subramanian V,et al.Anti-inflammatory effect of certain dimethoxy flavones[J].Inflammopharmacology,2015,23:307-317.