Xing Ying, Zhang Min, Wang Man-Man, Feng Ya-Shuo, Dong Fang, Zhang Feng
Department of Rehabilitation Medicine, The Third Hospital of Hebei Medical University, Shijiazhuang, China.
Department of Pathophysiology, Hebei Medical University, Shijiazhuang, China.
Front Cell Neurosci. 2021 Feb 2;15:633280. doi: 10.3389/fncel.2021.633280. eCollection 2021.
The main purpose of the study was to investigate the antiapoptotic effect of electroacupuncture (EA) in the acute stage of ischaemic stroke in rats. The cerebral ischemia model was established by middle cerebral artery occlusion (MCAO)/reperfusion in rats. A single EA treatment was performed at the acute stage of ischaemic stroke. The neurological function, brain water content, apoptotic cell number, and cerebral infarct volume were assessed in stroke rats. The expression of autophagy-related proteins (LC3II/I, Beclin1, P62, and LAMP1), Sirtuin 1 (SIRT1), p-JNK, p-ERK1/2, and cleaved caspase-3 (CCAS3) were measured by Western blot, immunofluorescence, and immunohistochemistry. Rapamycin (RAP, an activator of autophagy) was used to confirm the antiapoptotic effect of EA regulating autophagy. The brain edema infarct size and apoptotic cell number were increasing within 3 days following stroke, and brain edema reached its peak at 24 h after stroke. EA treatment at 24 h after ischaemic stroke obviously suppressed the number of apoptotic cells and brain edema. However, there were no significant differences in infarct volumes among EA-12 h, EA-24 h, and MCAO/R group. Moreover, EA treatment at 24 h after ischaemic stroke obviously suppressed the expression of CCAS3, LC3II/I, Beclin1 while increasing the level of P62 and LAMP1 and hence mediating autophagy, which was reversed by RAP. Meanwhile, the expression of SIRT1, p-ERK1/2, p-JNK were promoted by EA at 24 h after ischaemic stroke. In conclusion, EA treatment may suppress apoptosis possibly regulating autophagy in the acute period after ischaemic stroke, hence reducing brain injury.
本研究的主要目的是探讨电针(EA)对大鼠缺血性脑卒中急性期的抗凋亡作用。通过大脑中动脉闭塞(MCAO)/再灌注建立大鼠脑缺血模型。在缺血性脑卒中急性期进行单次EA治疗。评估脑卒中大鼠的神经功能、脑含水量、凋亡细胞数量和脑梗死体积。通过蛋白质免疫印迹法、免疫荧光法和免疫组织化学法检测自噬相关蛋白(LC3II/I、Beclin1、P62和LAMP1)、沉默调节蛋白1(SIRT1)、磷酸化JNK(p-JNK)、磷酸化细胞外信号调节激酶1/2(p-ERK1/2)和裂解的半胱天冬酶-3(CCAS3)的表达。使用雷帕霉素(RAP,一种自噬激活剂)来证实EA通过调节自噬的抗凋亡作用。脑卒中后3天内脑水肿梗死面积和凋亡细胞数量增加,脑水肿在脑卒中后24小时达到峰值。缺血性脑卒中后24小时进行EA治疗可明显抑制凋亡细胞数量和脑水肿。然而,EA-12小时组、EA-24小时组和MCAO/R组之间的梗死体积没有显著差异。此外,缺血性脑卒中后24小时进行EA治疗可明显抑制CCAS3、LC3II/I、Beclin1的表达,同时增加P62和LAMP1的水平,从而介导自噬,RAP可逆转这种作用。同时,缺血性脑卒中后24小时EA可促进SIRT1、p-ERK1/2、p-JNK的表达。总之,EA治疗可能通过调节缺血性脑卒中急性期的自噬来抑制细胞凋亡,从而减轻脑损伤。