Rehabilitation Medical College of Henan University of TCM, Zhengzhou 450046, China.
Second Affiliated Hospital of Henan University of TCM, Zhengzhou 450002.
Zhongguo Zhen Jiu. 2024 Feb 12;44(2):175-181. doi: 10.13703/j.0255-2930.20220818-k0008.
To investigate the effects of electroacupuncture (EA) on the miR-381, leucine-rich repeat C4 protein (LRRC4), and downstream stromal cell-derived factor-1 (SDF-1)/CXC chemokine receptor 4 (CXCR4) signaling pathway in rat model of ischemic stroke, and to explore the mechanism by which EA improves neurological damage following ischemic stroke.
Among 50 SPF male SD rats, 10 rats were randomly selected into a sham surgery group, and the remaining rats were used to establish the middle cerebral artery occlusion (MCAO) model. The 30 successfully modeled rats were randomly divided into a model group, an EA group, and an agonist group, with 10 rats in each group. The rats in the EA group received EA at "Baihui" (GV 20) and "Dazhui" (GV 14), with disperse-dense wave, a frequency of 2 Hz/10 Hz, and a current intensity of 1 mA, 30 min per session, once daily for a total of 14 days. The rats in the agonist group received miR-381 agonist injections into the lateral ventricle, with 10 μL per injection, every 7 days for a total of 2 injections. After intervention, ZeaLonga neurobehavioral deficit score was observed in each group. HE staining was performed to observe the morphological changes in the ischemic brain tissue of rats in each group. ELISA was used to measure the levels of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), and nerve growth factor (NGF) in serum. Western blot was employed to detect the protein expression of LRRC4, SDF-1, CXCR4, and extracellular regulated protein kinase 1 (ERK1) in the ischemic brain tissue. Real-time PCR was utilized to assess the expression of miR-381 and LRRC4, SDF-1, CXCR4, ERK1 mRNA in the ischemic brain tissue.
After intervention, the brain tissue showed disordered cell arrangement, reduced quantity, and significant interstitial edema, with numerous vacuoles in the model group. The pathological changes mentioned above were alleviated in the brain tissue of rats in the EA group and the agonist group. Compared with the sham surgery group, the rats in the model group exhibited increased ZeaLonga neurobehavioral deficit scores, elevated levels of serum TNF-α and IL-6 (<0.01), and decreased serum NGF level (<0.01)；the protein expression of SDF-1, CXCR4 and ERK1 in ischemic brain tissue was reduced (<0.01), while LRRC4 protein expression was increased (<0.01)；the expression of miR-381, as well as SDF-1, CXCR4 and ERK1 mRNA in ischemic brain tissue was decreased (<0.01), while LRRC4 mRNA expression was increased (<0.01). Compared with the model group, the rats in the EA group and the agonist group showed decreased ZeaLonga neurobehavioral deficit scores and reduced levels of serum TNF-α and IL-6 (<0.05, <0.01), and increased serum NGF levels (<0.05, <0.01); the protein expression of SDF-1, CXCR4 and ERK1 in ischemic brain tissue was increased (<0.01), while LRRC4 protein expression was decreased (<0.01)；the expression of miR-381, as well as SDF-1, CXCR4 and ERK1 mRNA in ischemic brain tissue was increased (<0.05, <0.01), while LRRC4 mRNA expression was decreased (<0.01).
EA at "Baihui" (GV 20) and "Dazhui" (GV 14) may promote the repair of neurological damage following ischemic stroke by up-regulating miR-381 to selectively inhibit LRRC4 expression, thereby activating the SDF-1/CXCR4 signaling pathway.
探讨电针对缺血性中风大鼠模型 miR-381、富含亮氨酸重复 C4 蛋白（LRRC4）和下游基质细胞衍生因子-1（SDF-1）/CXC 趋化因子受体 4（CXCR4）信号通路的影响，探讨电针改善缺血性中风后神经损伤的机制。
在 50 只 SPF 雄性 SD 大鼠中，随机选择 10 只大鼠进入假手术组，其余大鼠用于建立大脑中动脉闭塞（MCAO）模型。成功建模的 30 只大鼠随机分为模型组、电针组和激动剂组，每组 10 只。电针组大鼠取“百会”（GV 20）和“大椎”（GV 14）穴，疏密波，频率 2 Hz/10 Hz，电流强度 1 mA，每次 30 min，每日 1 次，共 14 天。激动剂组大鼠向侧脑室注射 miR-381 激动剂，每次 10 μL，每 7 天注射 1 次，共 2 次。干预后，观察各组大鼠的 ZeaLonga 神经行为学缺损评分。HE 染色观察各组大鼠缺血脑组织的形态学变化。ELISA 法检测血清肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）和神经生长因子（NGF）水平。Western blot 法检测缺血脑组织中 LRRC4、SDF-1、CXCR4 和细胞外调节蛋白激酶 1（ERK1）的蛋白表达。实时 PCR 法检测缺血脑组织中 miR-381 和 LRRC4、SDF-1、CXCR4、ERK1mRNA 的表达。
干预后，模型组脑组织细胞排列紊乱，数量减少，间质水肿明显，大量空泡。电针组和激动剂组大鼠脑组织的病理变化得到缓解。与假手术组相比，模型组大鼠的 ZeaLonga 神经行为学缺损评分升高（<0.01），血清 TNF-α和 IL-6 水平升高（<0.01），血清 NGF 水平降低（<0.01）；缺血脑组织 SDF-1、CXCR4 和 ERK1 蛋白表达减少（<0.01），LRRC4 蛋白表达增加（<0.01）；缺血脑组织 miR-381、SDF-1、CXCR4 和 ERK1mRNA 的表达减少（<0.01），LRRC4mRNA 的表达增加（<0.01）。与模型组相比，电针组和激动剂组大鼠的 ZeaLonga 神经行为学缺损评分降低（<0.05，<0.01），血清 TNF-α和 IL-6 水平降低（<0.05，<0.01），血清 NGF 水平升高（<0.05，<0.01）；缺血脑组织 SDF-1、CXCR4 和 ERK1 蛋白表达增加（<0.01），LRRC4 蛋白表达减少（<0.01）；缺血脑组织 miR-381、SDF-1、CXCR4 和 ERK1mRNA 的表达增加（<0.05，<0.01），LRRC4mRNA 的表达减少（<0.01）。
电针“百会”（GV 20）和“大椎”（GV 14）可能通过上调 miR-381 选择性抑制 LRRC4 表达，从而激活 SDF-1/CXCR4 信号通路，促进缺血性中风后神经损伤的修复。
