Objectives: To explore the possible mechanism of Shao's five-needle therapy pretreatment on relieving airway inflammatory response in asthmatic rats.
Methods: Forty SPF-grade SD rats were randomly divided into a blank group, a model group, an acupuncture group, and a medication group, with 10 rats in each group. Except the blank group, asthma model was established by aerosol inhalation of ovalbumin in the other 3 groups. The rats in the acupuncture group were treated with acupuncture at "Dazhui" (GV 14) and bilateral "Feishu" (BL 13) and "Fengmen" (BL 12), with each session lasting for 20 min. Acupuncture was given before each motivating, once daily for 7 consecutive days. The rats in the medication group were treated with intraperitoneal injection of dexamethasone sodium phosphate solution before each motivating, once daily for 7 days. General situation of the rats was observed in each group; ELISA method was used to detect the levels of inflammatory cytokines interleukin (IL)-1β and IL-18 in serum; immunofluorescence staining method was performed to assess the expression of reactive oxygen species (ROS) in lung tissues; Western blot method was used to measure the protein expression of thioredoxin interacting protein (TXNIP), NOD-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), and Caspase-1 in lung tissues.
Results: The rats in the blank group exhibited normal behavior, while those in the model group showed signs of respiratory distress, ear scratching, cheek rubbing, and dysphoria. Compared with the model group, the rats in the acupuncture group and the medication group showed stable respiration and relatively agile responses. Compared with those in the blank group, the serum levels of IL-18 and IL-1β were elevated (P<0.01), the expression intensity of ROS was increased, and the protein expressions of TXNIP, NLRP3, ASC and Caspase-1 in lung tissues were increased (P<0.01) in the model group. Compared with those in the model group, the serum levels of IL-18 and IL-1β were reduced (P<0.01), the expression intensity of ROS was lowered, and the protein expressions of TXNIP, NLRP3, ASC and Caspase-1 in lung tissues were reduced (P<0.01) in the acupuncture group and the medication group. Compared with the medication group, the protein expression of ASC in lung tissue was reduced in the acupuncture group (P<0.05).
Conclusions: Pretreatment of Shao's five-needle therapy could alleviate airway inflammatory response in asthmatic rats by reducing ROS levels and decreasing the aggregation and activation of pathway-related proteins in the ROS/TXNIP/NLRP3 pathway, ultimately leading to decreased secretion of IL-1β and IL-18. This mechanism may contribute to the effectiveness of Shao's five-needle therapy in preventing and treating asthma.
目的: 探讨“邵氏五针法”预处理缓解哮喘大鼠气道炎性反应可能的作用机制。方法: 将40只SPF级SD大鼠随机分为空白组、模型组、针刺组和西药组，每组10只。除空白组外，其余3组大鼠采用雾化吸入卵蛋白致敏及激发方法制备哮喘模型。针刺组取“大椎”及双侧“肺俞”“风门”行针刺预处理干预，每次20 min，起针后进行激发，每日1次，连续7 d；西药组予腹腔注射地塞米松磷酸钠溶液，用药后激发，每日1次，连续7 d。观察各组大鼠一般情况，ELISA法检测大鼠血清炎性因子白细胞介素（IL）-1β、IL-18含量，免疫荧光染色法检测大鼠肺组织活性氧簇（ROS）阳性表达，Western blot法检测大鼠肺组织硫氧还蛋白反应蛋白（TXNIP）、NOD样受体蛋白3（NLRP3）、凋亡相关斑点样蛋白（ASC）以及半胱氨酸天冬氨酸特异蛋白酶（Caspase-1）蛋白表达。结果: 空白组大鼠精神行为状态良好；模型组大鼠出现呼吸急促、抓耳挠腮、烦躁不安等情况；与模型组比较，针刺组、西药组大鼠呼吸较平稳，反应相对灵敏。与空白组比较，模型组大鼠血清IL-18、IL-1β含量升高（P<0.01），ROS阳性表达强度增高，肺组织TXNIP、NLRP3、ASC、Caspase-1蛋白表达升高（P<0.01）；与模型组比较，针刺组、西药组大鼠血清IL-18、IL-1β含量降低（P<0.01），ROS阳性表达强度降低，肺组织TXNIP、NLRP3、ASC、Caspase-1蛋白表达降低（P<0.01）。与西药组比较，针刺组大鼠肺组织ASC蛋白表达降低（P<0.05）。结论: “邵氏五针法”预处理可通过降低ROS，减少ROS/TXNIP/NLRP3通路相关蛋白的聚集与激活，进而减少下游IL-1β、IL-18炎性因子的分泌，改善哮喘大鼠气道炎性反应，这可能是“邵氏五针法”防治哮喘的作用机制之一。.
Keywords: Caspase-1; NOD-like receptor protein 3; Shao's five-needle therapy; acupuncture; asthma; inflammatory response; reactive oxygen species; thioredoxin interacting protein.