背景:帕金森病(PD)是中老年人常见的中枢神经退行性疾病。多巴胺能神经元的进行性变性和死亡导致多巴胺(DA)神经递质不足。针灸可以缓解神经元的老化。因此,研究电针对PD小鼠的神经保护作用尤为重要。
方法:腹膜内注射1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP,20mg/kg)建立PD小鼠模型,脂多糖(LPS)用于诱导小胶质细胞极化。西方印迹,末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL),Nissl染色和免疫组化检测神经元凋亡和损伤,PD小鼠α-syn表达和小胶质细胞积累。此外,采用酶联免疫吸附试验(ELISA)测定血清炎症因子水平。流式细胞仪检测Ca2+含量。使用异硫氰酸荧光素(FITC)标记方法来评估葡萄糖摄取。试剂盒用于检测葡萄糖和乳酸水平。
结果:MPTP诱导小鼠SN中DA神经元的选择性丢失,改变了Ca2+稳态,并诱导炎症反应。此外,维持Ca2稳态取决于瞬时受体电位通道1(TRPC1)的活性。EA治疗促进TRPC1表达,它对钠-葡萄糖协同转运蛋白1(SGLT1)具有负调节作用。在EA的作用下,TRPC1蛋白表达增加,Ca2+浓度升高,SGLT1的作用受到抑制,从而促进葡萄糖代谢,阻断磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/AKT)途径的激活,抑制小胶质细胞M1极化,缓解PD过程。
结论:EA促进TRPC1/Ca2+通路激活,抑制SGLT1介导的糖代谢调节和PI3K/AKT通路激活,抑制小胶质细胞M1极化,缓解PD。
BACKGROUND: Parkinson\'s disease (PD) is a common central neurodegenerative disease in middle-aged and elderly people. The progressive degeneration and death of dopaminergic neurons leads to insufficient dopamine (DA) neurotransmitters. Acupuncture and moxibustion can alleviate the aging of neurons. Therefore, studying the neuroprotective effects of electroacupuncture (EA) in PD mice is particularly important.
METHODS: Intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP, 20 mg/kg) was used to establish a PD mouse model, and lipopolysaccharide (LPS) was used to induce microglia polarization. Western blotting, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), Nissl staining and immunohistochemistry were used to detect neuronal apoptosis and injury, α-syn expression and microglial accumulation in PD mice. In addition, the levels of serum inflammatory factors were determined using enzyme-linked immunosorbent assay (ELISA). Flow cytometry was used to detect the Ca2+ content. The fluorescein isothiocyanate (FITC) labeling method was used to assess glucose uptake. A reagent kit was used to detect glucose and lactate levels.
RESULTS: MPTP induced the selective loss of DA neurons in the SN of mice, altered Ca2+ homeostasis, and induced an inflammatory response. In addition, maintaining Ca2+ homeostasis depends on the activity of transient receptor potential channel 1 (TRPC1). EA therapy promotes TRPC1 expression, which has a negative regulatory effect on sodium-glucose cotransporter 1 (
SGLT1). Under the action of EA, TRPC1 protein expression increased, Ca2+ concentrations increased, and the effect of
SGLT1 was inhibited, thereby facilitating glucose metabolism, blocking the activation of the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) pathway, restraining M1 polarization of microglia, and alleviating the PD process.
CONCLUSIONS: EA promotes TRPC1/Ca2+ pathway activation, inhibits
SGLT1-mediated regulation of glucose metabolism and PI3K/AKT pathway activation, inhibits microglial M1 polarization, and alleviates PD.