化浊解毒活血通络法对大鼠脑缺血再灌注损伤后神经细胞凋亡及P38MAPK信号通路的影响

发布时间：2018-06-10 16:09

本文选题：化浊解毒活血通络法 + 脑缺血再灌注损伤　；参考：《河北医科大学》2016年硕士论文

【摘要】：目的:本研究在大鼠脑缺血再灌注损伤模型的基础上,运用化浊解毒活血通络方进行干预,通过对神经功能缺损评分、脑组织病理形态改变、用药后脑梗死体积变化、P-P38MAPK与Caspase-3的观察,探讨该方法是否通过抑制P38MAPK信号通路来调控细胞凋亡,进一步阐明化浊解毒活血通络法对受损神经元产生保护作用的内在机制。方法:健康雄性SD大鼠54只,随机分为6组:分别是假手术组、模型组、化浊解毒活血通络方低剂量组(以下简称“中药低剂量组”)、化浊解毒活血通络方中剂量组(以下简称“中药中剂量组”)、化浊解毒活血通络方高剂量组(以下简称“中药高剂量组”)和阿司匹林组。采用改良线栓法制备脑缺血再灌注损伤模型,于造模后第2天给予相应药物治疗,假手术组和模型组给予等量生理盐水灌胃,中药各剂量组灌服相应浓度的化浊解毒活血通络方中药液,阿司匹林组给予阿司匹林混悬液灌胃。连续灌胃14d后对各组大鼠进行神经功能缺损评分,处死后断头,每组选取3只大鼠取脑,进行TTC染色,测量梗死范围。其余6只取右侧大脑半球分别进行HE染色,光镜下观察缺血区大脑皮质组织的形态变化。PCR法分别检测脑皮质区P-P38MAPK和Caspase-3 mRNA的表达。结果:1神经功能缺损评分:与假手术组比较,模型组神经功能缺损评分明显升高(P0.05);与模型组相比,中药低剂量组、中药中剂量组神经功能缺损无明显差异(P0.05),中药高剂量组、阿司匹林组神经功能缺损评分均降低,差异有统计学意义(P0.05);与中药低剂量组比较,中药中、高剂量组神经功能缺损评分均下降(P0.05);但中药高剂量组与阿司匹林组相比,无明显统计学差异(P0.05)。2脑组织病理形态学改变:假手术组大鼠大脑皮层区脑组织结构正常,细胞排列整齐紧密,无变性坏死细胞;模型组大鼠大脑皮层区可见明显缺血区,脑组织明显水肿,神经组织排列紊乱,细胞间隙明显增大,可见大量的细胞变性坏死;中药低剂量组脑组织形态表现与模型组相似;中药中剂量组、中药高剂量组及阿司匹林组缺血区变性坏死细胞较模型组减少;中药高剂量组及阿司匹林组可见少量神经坏死细胞,脑组织形态无明显差异。3ttc法检测大鼠脑梗死体积:与假手术组相比,各组均存在较明显的梗死灶,其中模型组与中药低剂量组梗死体积无明显差异(p0.05);中药中、高剂量组及阿司匹林组梗死体积与模型组相比,均有所减小(p0.05);中药中、低剂量组脑梗死体积减少均则不及阿司匹林组(p0.05),中药高剂量组与阿司匹林组相比脑梗死体积无明显变化(p0.05)。4pcr法检测脑组织中caspase-3的表达:与假手术组相比,模型组caspase-3的表达水平明显升高(p0.05);与模型组比较,各用药组caspase-3的表达水平均降低(p0.05);与中药低剂量组比较,中药中剂量、中药高剂量组及阿司匹林组caspase-3的表达降低显著(p0.05);且中药中、高剂量组与阿司匹林组比较无明显统计学差异(p0.05)。5pcr法检测脑组织中p-p38mapk的表达:与假手术组比较,模型组p-p38mapk的表达明显升高,差异有统计学意义(p0.05);与模型组相比,各用药组p-p38mapk的表达水平均降低(p0.05);与中药低剂量组比较,中药高剂量组及阿司匹林组p-p38mapk的表达显著降低(p0.05);而中药高剂量组的表达与阿司匹林组相当,无明显统计学差异(p0.05)。结论:1化浊解毒活血通络方能明显改善脑缺血再灌注损伤大鼠受损脑组织的病理形态,降低该模型大鼠神经功能缺损评分。2化浊解毒活血通络方能够减小脑缺血再灌注损伤大鼠脑梗死体积,缩小梗死范围。3化浊解毒活血通络方能够抑制caspase-3的阳性表达及磷酸化p38mapk蛋白的活性,从而减少神经细胞凋亡。4化浊解毒活血通络方对脑缺血再灌注损伤的保护机制可能与抑制细胞凋亡有关,且可能是通过p38mapk信号通路完成的。
[Abstract]:Objective: on the basis of the model of cerebral ischemia reperfusion injury in rats, the purpose of this study was to intervene with the prescription of chemical detoxification and activating blood dredging collaterals, through the observation of the score of nerve function defect, the pathological changes of brain tissue, the change of cerebral infarction volume after drug use, the observation of P-P38MAPK and Caspase-3, and to explore whether this method can be controlled by inhibiting the P38MAPK signaling pathway. The internal mechanism of cell apoptosis was further clarified. Methods: 54 healthy male SD rats were randomly divided into 6 groups: the sham operation group, the model group, the low-dose group (hereinafter referred to as "the low dose group of Chinese medicine") in the sham operation group (hereinafter referred to as "the low dose group of Chinese medicine"), the huoxioxiu Huoxue Tongluo recipe. The dose group (hereinafter referred to as "the Chinese medicine dose group"), the high dose group (hereinafter referred to as "the high dose group of Chinese medicine") and the aspirin group (hereinafter referred to as "the high dose group of Chinese medicine") and the aspirin group. The model of cerebral ischemia reperfusion injury was prepared by improved thread emboli method. The corresponding drug treatment was given second days after the model building, and the sham operation group and the model group were given the same amount of saline irrigation. The stomach, each dose group of Chinese medicine was given the corresponding concentration of the solution of the decoction of huoxioxiu Huoxue Tongluo, the aspirin group was given the aspirin mixed suspensions. After continuous perfusion of 14d, the nerve function defect was scored in each group. After the death, the brain was taken in each group of 3 rats and the TTC staining was performed to measure the infarct scope. The other 6 only took the right brain. The hemispheres were stained with HE, the morphological changes of cerebral cortex in the ischemic area were observed under light microscope and the expression of P-P38MAPK and Caspase-3 mRNA in the cerebral cortex were detected by.PCR method. Results: 1 the score of nerve function defect was compared with the sham group, the score of neural function defect in the model group was significantly higher (P0.05); compared with the model group, the low dose group of traditional Chinese medicine, There was no significant difference in nerve function defect in the medium dose group (P0.05). The scores of nerve function defect in the high dose group and aspirin group were all lower, the difference was statistically significant (P0.05). Compared with the low dose group of Chinese traditional medicine, the scores of nerve function defect in the high dose group decreased (P0.05), but the high dose group of traditional Chinese medicine was no more than the aspirin group. The pathological changes in the brain tissue of.2 were significantly different (P0.05): the cerebral cortex area of the rats in the sham operation group was normal, the cells arranged neatly and neatly, and the necrotic cells were arranged neatly. The cerebral cortex area of the rats in the model group showed obvious ischemic area, the brain tissue was edema, the nerve tissue was arranged in disorder, and the space between the cells was obviously enlarged, and a large number of them could be seen. The morphology of the brain tissue in the low dose group of Chinese medicine was similar to that in the model group, and the medium dose group, the high dose group and the aspirin group were less than the model group, and the high dose group and the aspirin group showed a small amount of nerve necrotic cells in the high dose group and the aspirin group, and the brain tissue morphology was not significantly different from the.3ttc method. Cerebral infarction volume: compared with the sham group, there were obvious infarcts in all groups, and there was no significant difference in infarct volume between the model group and the low dose group of traditional Chinese medicine (P0.05). The infarct volume in the high dose group and the aspirin group was lower than that in the model group (P0.05), and the decrease of cerebral infarction volume in the low dose group was less than that of the Chinese traditional medicine. Compared with the sham group, the expression level of Caspase-3 in the model group was significantly higher than that in the sham group (P0.05) and the expression level of Caspase-3 in the model group was significantly higher than that in the sham group (P0.05), and the expression level of Caspase-3 in each group was lower (P0.05) than that in the model group (P0.05), compared with the model group (P0.05), compared with the group of the sham operation group (P0.05). Compared with the low dose group, the expression of Caspase-3 in the high dose group and the aspirin group decreased significantly (P0.05), and there was no significant difference between the high dose group and the aspirin group in the traditional Chinese medicine (P0.05).5pcr method to detect the expression of P-P38MAPK in the brain tissue: the expression of P-P38MAPK in the model group was significantly higher than that in the sham operation group. The difference was statistically significant (P0.05). Compared with the model group, the expression level of P-P38MAPK in each group decreased (P0.05). Compared with the low dose group, the expression of P-P38MAPK in the high dose group and the aspirin group decreased significantly (P0.05), while the high dose group was similar to the aspirin group, and there was no significant statistical difference (P0.05). 1 huoxihou Jiedu Huoxue Tongluo recipe can obviously improve the pathological morphology of the damaged brain tissue of the rats with cerebral ischemia reperfusion injury, and reduce the nerve function defect score of the model rats,.2 turbidity, detoxification and activating blood circulation can reduce the volume of cerebral infarction in the rats with cerebellar ischemia reperfusion injury, and reduce the infarct scope to inhibit the CAS of the turbid and activating blood circulation and dredging collaterals. The positive expression of pase-3 and the activity of phosphorylated p38MAPK protein, thus reducing the protective mechanism of neuronal apoptosis,.4, turbid, activating blood circulation and collaterals on cerebral ischemia reperfusion injury, may be related to the inhibition of apoptosis, and may be accomplished through the p38MAPK signaling pathway.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R277.7

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