College of Acupuncture-Moxibustion and Tuina, Nanjing University of Chinese Medicine, Nanjing 210023, China; Key Laboratory of Acupuncture and Medicine Research of Ministry of Education, Nanjing University of Chinese Medicine, Nanjing 210023, China.
School of Chinese Medicine, Nanjing University of Chinese Medicine, Nanjing 210023, China.
Brain Res. 2024 Dec 15;1845:149296. doi: 10.1016/j.brainres.2024.149296. Epub 2024 Oct 28.
Thrombolytic therapy is the primary treatment for acute ischemic stroke. Extending the therapeutic time window can effectively reduce the harmful side effects associated with thrombolytic therapy. Although electroacupuncture (EA) has been shown to extend this time window, the specific mechanisms remain unclear.
We developed an embolic stroke model in rats and administered EA during thrombolytic therapy with recombinant tissue plasminogen activator (rt-PA) either 4.5 or 6 h after stroke onset. Neurological deficits were evaluated at 2 and 24 h post-stroke. Brain tissue was collected for analysis using 2,3,5-triphenyl tetrazolium chloride (TTC) staining, water content measurement, blood-brain barrier (BBB) permeability assessment, electron microscopy, and TUNEL assay. Immunofluorescence staining, western blotting, and enzyme-linked immunosorbent assays were employed to quantify the expression of proteins related to BBB integrity and pyroptosis.
Neuronal damage and BBB disruption along with increased expression of pyroptosis-related proteins were observed following thrombolytic therapy at the 6-hour mark. EA treatment improved neurological outcomes, reduced infarct volume, and alleviated BBB disruption. EA also inhibited the expression of matrix metalloproteinase 9 (MMP9) and enhanced the expression of tissue inhibitor of metalloproteinases 1 (TIMP1), helping to maintain BBB integrity. Furthermore, EA reduced the expression of pyroptosis-related proteins, including gasdermin D (GSDMD), interleukin-1β (IL-1β), and interleukin-18 (IL-18). EA also reduced the co-expression of GSDMD and MMP9 in brain tissues.
EA may be a promising therapeutic approach for extending the thrombolytic therapy window by protecting the BBB and inhibiting GSDMD-mediated pyroptosis.
溶栓治疗是急性缺血性脑卒中的主要治疗方法。延长治疗时间窗可以有效降低溶栓治疗相关的有害副作用。虽然电针（EA）已被证明可以延长这个时间窗，但具体机制尚不清楚。
我们在大鼠中建立了栓塞性脑卒中模型，并在脑卒中发作后 4.5 或 6 小时给予重组组织型纤溶酶原激活物（rt-PA）溶栓治疗时进行 EA。在脑卒中后 2 和 24 小时评估神经功能缺损。使用 2,3,5-三苯基氯化四氮唑（TTC）染色、水含量测量、血脑屏障（BBB）通透性评估、电子显微镜和 TUNEL 测定分析脑组织。免疫荧光染色、Western blot 和酶联免疫吸附测定用于定量与 BBB 完整性和细胞焦亡相关的蛋白表达。
在 6 小时溶栓治疗后，观察到神经元损伤和 BBB 破坏以及细胞焦亡相关蛋白表达增加。EA 治疗改善了神经功能预后，减少了梗死体积，缓解了 BBB 破坏。EA 还抑制了基质金属蛋白酶 9（MMP9）的表达，增强了组织金属蛋白酶抑制剂 1（TIMP1）的表达，有助于维持 BBB 完整性。此外，EA 降低了细胞焦亡相关蛋白的表达，包括 Gasdermin D（GSDMD）、白细胞介素-1β（IL-1β）和白细胞介素-18（IL-18）。EA 还减少了脑组织中 GSDMD 和 MMP9 的共表达。
EA 可能是一种有前途的治疗方法，可以通过保护 BBB 和抑制 GSDMD 介导的细胞焦亡来延长溶栓治疗时间窗。
