目的: 探讨CXCL3是否通过介导炎症反应参与脑缺血再灌注损伤(cerebral ischemia-reperfusion injury, CIRI)。方法: 将SD雄性大鼠随机分为假手术组、模型组、CXCL3注射组(200 ng/只)。假手术组仅进行手术暴露血管;模型组采用Longa线栓法建立大脑中动脉阻塞/再灌注(middle cerebral artery occlusion/reperfesion, MCAO/R)模型,其中缺血1 h再灌注24 h;CXCL3注射组采用脑立体定位法向MCAO/R模型侧脑室注射重组CXC趋化因子配体3(CXC chemokine ligand 3, CXCL3)蛋白。采用moorFLPI-2激光散斑血流成像仪检测MCAO模型局部脑血流的变化,神经功能评分法检测大鼠神经功能损伤程度,TTC染色法检测脑梗死体积,蛋白免疫印迹法(western blotting, WB)检测CXCL3以及白细胞介素-1(interleukin-1, IL-1)、白细胞介素-6(interleukin-6, IL-6)的蛋白表达情况。结果: 与基线值血流相比,MCAO模型大鼠血流降幅约70%(P＜0.001);与缺血1 h相比,拔线后血流值明显上升(P＜0.001),与基线值血流相比差异无统计学意义(P＞0.05)。再灌注24 h后,与假手术组相比,模型组大鼠的神经功能评分、脑梗死体积均增加(P＜0.001)。相较于假手术组,模型组缺血区CXCL3蛋白的表达上调(P＜0.001),炎症因子IL-1和IL-6蛋白表达均有所增加(P＜0.05);相较于模型组,注射重组CXCL3蛋白能够使CXCL3蛋白的表达进一步上调(P＜0.05),炎症因子IL-1和IL-6的表达水平均进一步上升(P＜0.05)。结论: 高水平的CXCL3可能通过上调促炎因子表达参与CIRI。
Objective: To investigate whether CXCL3 is involved in cerebral ischemia-reperfusion injury (CIRI) by mediating inflammatory response. Methods: The SD male rats were randomly divided into three groups: the sham operation group, the model group, and the CXCL3 injection group (200 ng per rat). The sham surgery group only underwent surgical exposure of the blood vessels. The model group established a middle cerebral artery occlusion/reperfusion (MCAO/R) model using the Longa suture method, with 1 hour of ischemia followed by 24 hours of reperfusion. The CXCL3 injection group used stereotactic brain surgery to inject recombinant CXC chemokine ligand 3 (CXCL3) protein into the lateral ventricle on the side of the MCAO/R model. The local cerebral blood flow changes in the MCAO model were detected using the moorFLPI-2 laser speckle perfusion imager. The degree of neurological dysfunction in rats was assessed using a neurological function scoring system. The volume of cerebral infarction was measured by TTC staining. The protein expression levels of CXCL3, interleukin-1 (IL-1) and interleukin-6 (IL-6), were measured by western blotting (WB). Results: Compared with the baseline blood flow, the blood flow in the MCAO model rats decreased by approximately 70% (P＜0.001). Compared with the blood flow after 1 hour of ischemia, the blood flow values significantly increased after suture was removed (P＜0.001). Additionally, there was no significant difference compared with the baseline blood flow (P＞0.05). Twenty-four hours after reperfusion, compared with the sham operation group, the neurological function score and cerebral infarction volume in the model group both significantly increased (P＜0.001). Compared with the sham operation group, the expression of CXCL3 protein in the ischemic area of the model group was upregulated (P＜0.001), and the expressions of the inflammatory factors IL-1 and IL-6 were both increased (P＜0.05). Compared with the model group, injection of recombinant CXCL3 protein further upregulate the expression of CXCL3 protein (P＜0.05), and significantly increased the expression levels of the inflammatory factors IL-1 and IL-6 (P＜0.05). Conclusion: High level of CXCL3 may contribute to CIRI by upregulating the expression of pro-inflammatory factors.
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