川芎嗪抑制大鼠脑缺血再灌注损伤的机制

发布时间：2018-02-05 20:38

本文关键词： 川芎嗪脑缺血再灌注损伤凋亡炎症　出处：《中国老年学杂志》2017年07期 　论文类型：期刊论文

【摘要】：目的探讨川芎嗪(TMP)抑制脑缺血再灌注损伤(CIRI)的分子机制。方法选择清洁级健康成年雄性SD大鼠,采用改良的Zea Longa栓线法构建大脑中动脉梗塞(MCAO)模型,并将大鼠随机分为三组(n=20):假手术组、模型组、TMP治疗组。假手术组大鼠不插入线栓,其余操作均同模型组;TMP治疗组大鼠在被拔出线栓再灌注时,腹腔注射TMP 20 mg/kg。再灌注24 h后,各组大鼠进行神经功能评分,然后麻醉处死,取材检测。干湿称重法检测大鼠脑组织含水量;2,3,5-三苯基氮化四氮唑(TTC)染色法检测大鼠脑梗死面积,HE染色观察脑组织形态;荧光定量PCR检测大鼠梗死侧脑皮层中miR-199a-5p、Bax、Bcl-2、肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β、IL-6 mRNA的表达水平;酶联免疫法检测大鼠血清中炎症介质TNF-α、IL-1β和IL-6的含量;免疫组化法观察大鼠梗死侧脑皮质核转录因子(NF)-κB p65及磷酸化NF-κB p65(NF-κB p-p65)的表达。结果 TMP治疗可显著改善神经缺陷评分(P0.05),降低脑组织含水量(P0.05),减少梗死体积(P0.01),减轻脑组织破坏。模型组miR-199a-5p、Bax、TNF-α、IL-1β和IL-6 mRNA的表达明显升高(P0.05),而Bcl-2 mRNA表达水平显著降低(P0.05)。而TMP治疗可显著降低miR-199a-5p、Bax、TNF-α、IL-1β和IL-6 mRNA的表达水平(P0.05),增加Bcl-2 mRNA的表达(P0.05)。与模型组比较,TMP治疗组大鼠脑皮质和血清中TNF-α、IL-1β和IL-6炎症因子含量均显著降低(P0.05)。IHC结果显示,TMP治疗组中NF-κB p-p65阳性细胞数明显减少(P0.05)。结论 TMP可减轻CIRI,其机制可能为抑制miR-199a-5p的表达,减少凋亡,降低炎症反应,且可降低NF-κB的活化。
[Abstract]:Objective to investigate the molecular mechanism of ligustrazine (TMP) inhibiting cerebral ischemia-reperfusion injury (CIRI). Methods healthy male SD rats of clean grade were selected. The model of middle cerebral artery infarction (MCAO) was established by modified Zea Longa method. The rats were randomly divided into three groups: sham operation group and model group. TMP treatment group, sham-operation group rats did not insert thread embolus, the rest of the operation is the same as the model group; The rats in the TMP group were injected intraperitoneally with TMP 20 mg / kg during reperfusion. After 24 hours of reperfusion, the rats in each group were evaluated for neurological function, and then were anesthetized and killed. The water content of brain tissue was measured by wet and dry weighing method. (2) the infarct size of rat brain was detected by TTC- staining method, and the morphology of brain tissue was observed by HE staining. Fluorescence quantitative PCR was used to detect miR-199a-5 pAX Bax-2, TNF- 伪 and IL-1- 尾 in the cerebral cortex of rats. The expression level of IL-6 mRNA; The levels of TNF- 伪 IL-6 and TNF- 伪 IL-1 尾 in rat serum were detected by enzyme-linked immunosorbent assay (Elisa). Immunohistochemical study of NF 魏 B p65 and phosphorylated NF- 魏 B p65 and NF- 魏 B p-p65 in the cerebral cortex of rats. Results TMP could significantly improve the neurological defect score (P0.05). In the model group, the brain tissue water content was reduced, the infarct volume was reduced, the infarct volume was reduced, and the brain tissue damage was alleviated. The model group was miR-199a-5pAXTNF- 伪. The expression of IL-1 尾 and IL-6 mRNA increased significantly (P 0.05). However, the expression level of Bcl-2 mRNA decreased significantly, while TMP treatment significantly decreased the expression of miR-199a-5 pAXN TNF- 伪. The expression level of IL-1 尾 and IL-6 mRNA was increased by P0.05, and the expression of Bcl-2 mRNA was increased compared with the model group. In TMP treatment group, the levels of IL-1 尾 and IL-6 in the cerebral cortex and serum of rats were significantly decreased (P 0.05). IHC results showed that TNF- 伪 and TNF- 伪 in the serum of rats were significantly lower than those in the control group (P < 0.05). In TMP treatment group, the number of NF- 魏 B p-p65 positive cells was significantly decreased (P 0.05). Conclusion TMP can alleviate CIRI. Its mechanism may be to inhibit the expression of miR-199a-5p, reduce apoptosis, reduce inflammatory response, and reduce the activation of NF- 魏 B.
【作者单位】：南京中医药大学附属医院检验科;南京中医药大学第二临床学院;南京中医药大学基础医学院;
【基金】：国家自然科学基金资助课题(81403136,81373748,81171659,11574156)
【分类号】：R285.5

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