目的 深入了解丹参酮在脑梗中的影响和作用机制。方法 通过将大脑中动脉堵塞来制作脑梗大鼠模型。经丹参酮预处理后，对脑梗体积、脑水肿和神经功能缺损予以评价并使用TUNEL检查海马体和大脑皮层中的细胞凋亡情况，再利用酶联免疫吸附剂测定来确定白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）和 C-反应蛋白（CRP）的含量水平。另外在缺氧缺糖（OGD）条件下将大鼠原代神经细胞分离并进行培养。经丹参酮预处理后，分别通过MTT测定和流式细胞分析来观察细胞存活和细胞凋亡情况。Bax和Bcl-2的基因表达通过qRT-PCR and 蛋白质免疫印迹予以检测。结果 和无任何治疗处理的脑梗大鼠相比，丹参酮治疗可显著降低脑梗体积、脑水肿和神经功能缺损评分（P<0.05）。经丹参酮预处理后，细胞凋亡以及脑梗大鼠海马体和大脑皮层中IL-6、TNF-α和CRP 的含量水平均得到有效抑制（P<0.05）。此外，在缺氧缺糖诱发的鼠神经元细胞中，丹参酮还能显著增加细胞活性并抑制细胞凋亡比（P<0.05）。与此同时，丹参酮治疗可显著下调促凋亡分子Bax的基因表达并上调抗凋亡分子Bcl-2的基因表达（P<0.05）。结论 丹参酮通过抑制神经元细胞凋亡和体外体内炎症反应而发挥脑梗预防作用。
Objective To investigate the effect and mechanisms of tanshinone in cerebral infarction. Methods The cerebral infarction rat model was established by middle cerebral artery occlusion. After pretreatment with tanshinone, cerebral infarct volume, cerebral edema and neurological deficits score were evaluated, as well as cell apoptosis in hippocampus and cortex of the brain was examined with TUNEL and the levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and C-reactive protein (CRP) were determined by enzyme-linked immuno sorbent assay. In addition, rat primary neuronal cells were isolated and cultured in oxygen-glucose deprivation (OGD) conditions. After pretreatment with tanshinone, cell viability and apoptosis were observed by MTT assay and flow cytometry analysis, respectively. The expressions of Bax and Bcl-2 were detected by qRT-PCR and western blotting. Results Compared with untreated cerebral infarction rat, tanshinone treatment significantly reduced cerebral infarct volume, cerebral edema and neurological deficits score (P<0.05). Cell apoptosis as well as the levels of IL-6, TNF-α and CRP in hippocampus and cortex of cerebral infarction rat were inhibited after pretreatment with tanshinone (P<0.05). Furthermore, tanshinone remarkably increased cell viability and inhibited cell apoptosis ratio (P<0.05) in oxygen-glucose deprivation induced mouse neuronal cells. Meanwhile, tanshinone treatment significantly down-regulated the expression of pro-apoptotic molecule Bax and up-regulated the expression of anti-apoptotic molecule Bcl-2 (P<0.05). Conclusion Tanshinone has a preventive effect on cerebral infarction by inhibiting neuronal cell apoptosis and inflammatory response in vitro and in vivo.
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