摘要 目的：线粒体功能障碍与缺血性脑卒中的发生有着密切联系。同时，循经按摩能够促进脑卒中患者恢复神经元功能。因此，本研究深入探讨了循经按摩对缺血性脑卒中大鼠脑组织线粒体自噬的作用效果，为开发针对缺血性脑卒中疾病的有效治疗策略提供理论依据。 方法：构建缺血性脑卒中大鼠模型，将建模成功的缺血性脑卒中大鼠均分为模型组、按摩组、通路激动剂组、通路抑制剂组以及按摩＋抑制剂组，空白对照组大鼠采用常规按摩，按摩组采用循经按摩，通路激动剂组采用HIF-1α/BNIP3激活剂进行干预，通路抑制剂组采用HIF-1α/BNIP3抑制剂进行干预，按摩＋抑制剂组采用循经按摩和HIF-1α/BNIP3抑制剂进行联合干预，另选10只健康大鼠进行假手术干预。观察各组小鼠神经功能缺损、脑梗死体积、脑半球肿胀的指标，采用HE染色、透射电镜方法观察海马组织的病理变化和海马区域自噬小体的数量，并利用免疫荧光、蛋白免疫印迹实验检测大鼠脑组织中HIF-1α以及BNIP3的表达和脑组织线粒体中自噬（LC3B）、凋亡（Beclin-1）和HIF-1α/BNIP3信号通路相关蛋白表达。结果：（1）模型组海马组织相较于假手术组中HIF-1α以及BNIP3荧光表达强度增强（P<0.05）；按摩组海马组织相较于模型组中HIF-1α以及BNIP3荧光表达强度显著增强（P<0.05）；（2）与模型组相比，抑制剂组脑半球肿胀、神经功能缺损评分以及脑梗死体积水平上升（P<0.05）；激动剂组脑半球肿胀、神经功能缺损评分以及脑梗死体积水平下降（P<0.05）；激动剂组Beclin-1以及LC3B蛋白表达上升（P<0.05）；抑制剂组Beclin-1以及LC3B蛋白表达下降（P<0.05）；（3）与模型组相比，按摩组脑半球肿胀、神经功能缺损评分以及脑梗死体积下降（P<0.05），海马区细胞排列整齐，层次较为清晰，细胞数量较多，受损程度减轻，结构较完整；与按摩组相比，按摩+抑制剂组海马区细胞排列较为分散，细胞数量变少，出现水肿现象，组织呈现受损状态。（4）与模型组相比，按摩组LC3B、HIF-1α、Beclin-1以及BNIP3蛋白表达上升（P<0.05）；按摩＋抑制剂组LC3B、HIF-1α、Beclin-1以及BNIP3蛋白表达下降（P<0.05）。结论：采用循经按摩治疗缺血性脑卒中时能通过激活HIF-1α/BNIP3信号通路，进而促进脑组织线粒体自噬改善神经功能，为后续循经按摩在缺血性脑卒中患者临床治疗中的应用提供了理论基础。

Abstract Objective: Mitochondrial dysfunction is closely related to the occurrence of ischemic stroke. At the same time, meridian massage can promote the recovery of neuronal function in stroke patients. Therefore, this study further explored the effect of meridian massage on mitochondrial autophagy in brain tissue of ischemic stroke rats, providing theoretical basis for the development of effective treatment strategies for ischemic stroke diseases.Methods: The ischemic stroke rat model was constructed, and the ischemic stroke rats successfully modeled were divided into model group, massage group, pathway agonist group, pathway inhibitor group and massage + inhibitor group. The rats in the blank control group received conventional massage, the massage group received menstrual cycle massage, and the pathway agonist group received HIF-1α/BNIP3 activator intervention. The pathway inhibitor group was treated with HIF-1α/BNIP3 inhibitor, and the massage + inhibitor group was treated with the combination of menstrual massage and HIF-1α/BNIP3 inhibitor, and 10 healthy rats were selected for sham operation intervention. The indexes of nerve function deficit, cerebral infarction volume and cerebral hemisphere swelling were observed. The pathological changes of hippocampus and the number of autophagosomes in hippocampus were observed by HE staining and transmission electron microscopy. The expressions of HIF-1α and BNIP3 in brain tissue, autophagy (LC3B), apoptosis (Beclin-1) and HIF-1α/BNIP3 signaling pathways in brain mitochondria were detected by immunofluorescence and western blot assay.Results: (1) Compared with sham operation group, HIF-1α and BNIP3 fluorescence expression intensity were increased in model group (P<0.05); The fluorescence expression intensity of HIF-1α and BNIP3 in hippocampus of massage group was significantly increased compared with that of model group (P<0.05). (2) Compared with model group, cerebral hemisphere swelling, nerve function deficit score and cerebral infarction volume in inhibitor group were increased (P<0.05); The levels of cerebral hemisphere swelling, nerve function deficit score and cerebral infarction volume in agonist group were decreased (P<0.05). The protein expressions of Beclin-1 and LC3B in agonist group were increased (P<0.05). The protein expressions of Beclin-1 and LC3B in inhibitor group were decreased (P<0.05). (3) Compared with model group, cerebral hemisphere swelling, nerve function deficit score and cerebral infarction volume in massage group were decreased (P<0.05), hippocampal cells were arranged neatly, the hierarchy was clearer, the number of cells was more, the damage degree was reduced, and the structure was more complete; Compared with the massage group, the hippocampal cells in the massage + inhibitor group were more dispersed, the number of cells decreased, edema appeared, and tissue was damaged. (4) Compared with model group, the protein expressions of LC3B, HIF-1α, Beclin-1 and BNIP3 in massage group were increased (P<0.05); The protein expressions of LC3B, HIF-1α, Beclin-1 and BNIP3 in massage + inhibitor group were decreased (P<0.05). Conclusion: In the treatment of ischemic stroke, meridian massage can activate HIF-1α/BNIP3 signaling pathway, and then promote brain mitochondrial autophagy to improve nerve function, which provides a theoretical basis for the subsequent application of meridian massage in the clinical treatment of ischemic stroke patients.

湖北省自然科学基金项目任务（编号;2023AFD170）