College of Acupuncture-moxibustion and Orthopaedics of Hubei University of Chinese Medicine/Hubei Provincial Collaborative Innovation Center of Preventive Treatment of Diseases by Acupuncture-moxibustion, Wuhan 430060, China.
Zhen Ci Yan Jiu. 2024 Jan 25;49(1):15-22. doi: 10.13702/j.1000-0607.20230407.
To observe the effect of electroacupuncture (EA) on nuclear transcription factor E2 related factor 2 (Nrf2)/NOD-like receptor family pyrin domain-containing protein 3 (NLRP3)/cysteine aspartic acid specific protease-1 (Caspase-1) pathway in the substantia nigra (SN) of mice with Parkinson's disease (PD), so as to explore the neuroprotective mechanism of EA.
Forty C57BL/6 male mice were randomly divided into 4 groups, namely, control, PD model, EA and sham-EA groups, with 10 mice in each group. The PD mouse model was established by gavage of rotenone for 4 weeks. Mice in the EA group were given EA stimulation (1 mA, 2 Hz) at "Fengfu" (GV36), bilateral "Taichong" (LR3) and "Zusanli" (ST36) for 30 min, once daily for 2 consecutive weeks. And mice in the sham-EA group were given acupuncture at the subcutaneous areas of the same acupoints without EA stimulation. The open-field test was used for assessment of mouse behavior. The levels of interleukin-1β (IL-1β) and interleukin-18 (IL-18) in the serum were detected by enzyme-linked immunosorbent assay . The positive expression of tyrosine hydroxylase (TH) in SN was determined by immunohistochemistry. The mRNA expression levels of Nrf2, NLRP3, Caspase-1, gasdermin D(GSDMD), IL-1β, IL-18 and the protein expression levels of Nrf2, NLRP3, Caspase-1 and GSDMD in the SN were detected by quantitative real-time PCR and Western blot, separately.
After modeling, compared with the control group, the behavioral score was increased (<0.01), the total exercise time, the total distance and the average speed were decreased (<0.01), and the positive expression of TH and the mRNA and protein expression levels of Nrf2 in the SN were decreased (<0.01), while the contents of IL-1β and IL-18 in serum, the mRNA and protein expression levels of NLRP3, Caspase-1 and GSDMD and the mRNA expression levels of IL-1β and IL-18 in the SN were up-regulated (<0.01) in the PD model group. Following EA intervention, the behavioral score was decreased(<0.01), the total exercise time, total distance and average speed were increased (<0.01), the positive expression of TH and the mRNA and protein expressions of Nrf2 in SN were up-regulated (<0.01, <0.05), while the contents of IL-1β and IL-18 in serum, the mRNA and protein expression levels of NLRP3, Caspase-1 and GSDMD as well as the mRNA expression levels of IL-1β and IL-18 in the SN were down-regulated (<0.01, <0.05) in the EA group relative to the PD model and sham-EA groups. There were no significant differences in the above indicators between the PD model and sham-EA groups.
EA stimulation of GV36, LR3 and ST36 can improve motor deficits, reduce the loss of dopamine neurons in the SN, and inhibit neuroinflammatory responses in mice with PD, which may be related to its effects in regulating the Nrf2/NLRP3/Caspase-1 pathway mediated pyroptosis.
观察电针对帕金森病（PD）模型小鼠黑质（SN）核转录因子 E2 相关因子 2（Nrf2）/NOD 样受体家族富含亮氨酸重复域蛋白 3（NLRP3）/半胱天冬氨酸特异性蛋白酶-1（Caspase-1）通路的影响，探讨电针的神经保护机制。
40 只 C57BL/6 雄性小鼠随机分为对照组、PD 模型组、电针组和假电针组，每组 10 只。采用灌胃鱼藤酮的方法建立 PD 模型，持续 4 周。电针组给予电针刺激（1 mA、2 Hz），穴位为“风府”（GV36）、双侧“太冲”（LR3）和“足三里”（ST36），每天 1 次，连续 2 周。假电针组在相同的穴位皮下给予电针刺激。采用旷场实验评估小鼠行为。酶联免疫吸附试验检测血清中白细胞介素-1β（IL-1β）和白细胞介素-18（IL-18）的水平。免疫组织化学法检测 SN 中酪氨酸羟化酶（TH）的阳性表达。实时荧光定量 PCR 和 Western blot 分别检测 SN 中 Nrf2、NLRP3、Caspase-1、gasdermin D（GSDMD）、IL-1β、IL-18 的 mRNA 表达水平和 Nrf2、NLRP3、Caspase-1、GSDMD 的蛋白表达水平。
建模后，与对照组相比，PD 模型组行为评分升高（<0.01），总运动时间、总距离和平均速度降低（<0.01），SN 中 TH 阳性表达和 Nrf2 的 mRNA 和蛋白表达水平降低（<0.01），血清中 IL-1β和 IL-18 含量、SN 中 NLRP3、Caspase-1 和 GSDMD 的 mRNA 和蛋白表达水平以及 IL-1β和 IL-18 的 mRNA 表达水平升高（<0.01）。电针干预后，行为评分降低（<0.01），总运动时间、总距离和平均速度增加（<0.01），SN 中 TH 阳性表达和 Nrf2 的 mRNA 和蛋白表达水平升高（<0.01，<0.05），血清中 IL-1β和 IL-18 含量、SN 中 NLRP3、Caspase-1 和 GSDMD 的 mRNA 和蛋白表达水平以及 IL-1β和 IL-18 的 mRNA 表达水平降低（<0.01，<0.05）。PD 模型组和假电针组各指标差异无统计学意义。
电针刺激 GV36、LR3 和 ST36 可改善 PD 模型小鼠的运动障碍，减少 SN 中多巴胺能神经元的丢失，并抑制 PD 小鼠的神经炎症反应，这可能与其调节 Nrf2/NLRP3/Caspase-1 通路介导的细胞焦亡有关。
