摘要
目的:观察电针足三里穴和肺俞穴是否通过NLRP3/caspase-1通路介导的细胞焦亡途径减轻脓毒症诱导的大鼠急性肺损伤。方法:成年健康雄性SD大鼠30只,采用随机数字表法分为5组:对照组、模型组、电针组、电针+NLRP3激活剂组、非经非穴电针组,每组6只。采用盲肠结扎穿孔法建立脓毒症模型。电针组选取双侧足三里穴和肺俞穴给予疏密波电针刺激(模型制备前5 d每天进行1次,模型制备过程中每6 h进行1次,30 min/次);非经非穴电针组以相同的参数电针刺激足三里穴和肺俞穴旁开5 mm非经非穴处。于模型制备前30 min电针+NLRP3激活剂组腹腔注射100 mg/kg尼日利亚菌素钠盐。建模后24 h时处死大鼠取肺组织,计算肺湿重/干重(W/D)比值,观察病理学结果并行肺损伤评分;采用ELISA法测定肺组织白细胞介素(IL)-1β、IL-18含量;采用Western blot法检测肺组织核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)、凋亡相关斑点样蛋白(ASC)、裂解的半胱氨酰天冬氨酸特异性蛋白酶-1(cleaved-caspase-1)、N端消皮素-D(GSDMD-N)蛋白的表达水平。结果:与对照组比较,模型组、电针组、电针+NLRP3激活剂组和非经非穴电针组肺损伤评分、W/D比值、IL-1β含量及IL-18含量均升高,肺组织NLRP3、ASC、cleaved-caspase-1及GSDMD-N蛋白表达均上调,差异有统计学意义(P<0.05);与模型组比较,电针组肺损伤评分、W/D比值、IL-1β含量及IL-18含量均降低,肺组织NLRP3、ASC、cleaved-caspase-1及GSDMD-N蛋白表达均下调,差异有统计学意义(P<0.05);与电针组比较,电针+NLRP3激活剂组肺损伤评分、W/D比值、IL-1β含量及IL-18含量均升高,肺组织NLRP3、ASC、cleaved-caspase-1及GSDMD-N蛋白表达均上调,差异有统计学意义(P<0.05)。结论:电针刺激足三里穴和肺俞穴减轻大鼠脓毒症急性肺损伤的机制可能与抑制NLRP3/caspase-1通路介导的经典细胞焦亡途径有关。
Objective To observe whether electroacupuncture at Zusanli(ST36)and Feishu(BL13)acupoints reduces sepsis induced acute lung injury in rats through the pyroptosis pathway mediated by NLRP3/caspase-1 pathway.Methods Thirty adult healthy male SD rats were randomly divided into five groups:control group,model group,electroacupuncture group,electroacupuncture plus NLRP3 activator group,and non-acupoint electroacupuncture group,with six rats in each group.The sepsis model was established by cecal ligation and puncture.In the electroacupuncture group,bilateral ST36 and BL13 acupoints were stimulated with dense and sparse wave electroacupuncture(once daily for 5 days before model preparation,once every 6 hours during model preparation,30 minutes each time).In the non-acupoint electroacupuncture group,the same parameters of electroacupuncture were applied at non-acupoints located 5 mm away from ST36 and BL13.The electroacupuncture plus NLRP3 activator group received intraperitoneal injection of 100 mg/kg nigericin sodium salt 30 minutes before model preparation.After modeling,the rats were sacrificed at 24 hours,and lung tissues were collected to calculate the wet-to-dry weight ratio(W/D ratio),observe pathological changes,and evaluate lung injury scores.The levels of IL-1β,IL-18 in lung tissues were measured by ELISA,and the expression levels of NLRP3,ASC,cleaved caspase-1 and GSDMD-N proteins in lung tissues were detected by Western blot.Results Compared with the control group,the lung injury scores,W/D ratio,IL-1βand IL-18 levels in the model group,electroacupuncture group,electroacupuncture plus NLRP3 activator group,and non-acupoint electroacupuncture group were increased.The expressions of NLRP3,ASC,cleaved caspase-1,and GSDMD-N proteins in lung tissues were upregulated,and the differences were statistically significant(P<0.05).Compared with the model group,the lung injury scores,W/D ratio,IL-1βand IL-18 levels in the electroacupuncture group were decreased.The expressions of NLRP3,ASC,cleaved caspase-1,and GSDMD-N proteins in lung tissues were downregulated,and the differences were statistically significant(P<0.05).Compared with the electroacupuncture group,the lung injury scores,W/D ratio,IL-1βand IL-18 levels in the electroacupuncture plus NLRP3 activator group were increased.The expressions of NLRP3,ASC,cleaved caspase-1,and GSDMD-N proteins in lung tissues were upregulated,and the differences were statistically significant(P<0.05).Conclusion Electroacupuncture stimulation at ST36 and BL13 acupoints may alleviate acute lung injury in rats with sepsis through inhibiting the classical cell pyroptosis pathway mediated by NLRP3/caspase-1 pathway.
作者
张昊
苏乾
史佳
宫丽荣
ZHANG Hao;SU Qian;SHI Jia(Department of Anesthesiology and Critical Care Medicine,Tianjin Nankai Hospital,Tianjin Medical University,Tianjin 300100,China)
出处
《中国中西医结合外科杂志》
CAS
2023年第6期814-818,共5页
Chinese Journal of Surgery of Integrated Traditional and Western Medicine
基金
国家自然科学基金青年项目(82004076)。
关键词
电刺激疗法
肺损伤
细胞焦亡
Electrical acupoint stimulation
lung injury
pyroptosis
