经方调控NLRP3/Caspase-1通路对心房颤动大鼠炎症反应的影响*

作者：付向婷1，赵一慧1，王婉盈1，张文宗1,2

单位：1.河南中医药大学第二临床医学院，河南 郑州 450002；2.河南省中医院，河南 郑州 450002

引用：引用：付向婷，赵一慧，王婉盈，张文宗.经方调控NLRP3/Caspase-1通路对心房颤动大鼠炎症反应的影响[J].中医药导报,2025,31(11):31-37.

DOI：10.13862/j.cn43-1446/r.2025.11.006

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摘要：

目的：比较经方炙甘草汤、桂枝甘草汤、真武汤及苓桂术甘汤对大鼠心房颤动（简称房颤）发生的影响，筛选出效果最佳经方，并探讨效果最佳经方调控核苷酸结合结构域富含亮氨酸重复序列和含热蛋白结构域受体3（NLRP3）/胱天蛋白酶-1（Caspase-1）通路对心房颤动大鼠炎症反应的影响。方法：将42只雄性SD大鼠为随机分为空白组、模型组、炙甘草汤组、桂枝甘草汤组、真武汤组、苓桂术甘汤组及维拉帕米组，每组6只。空白组尾静脉注射生理盐水，其6组通过连续7 d尾静脉注射乙酰胆碱（Acetylchoine，Ach）-氯化钙（CaCl2）构建房颤模型。造模成功后各组予相应药物灌胃14 d。心电图监测各组大鼠房颤诱发时间与房颤持续时间，苏木精-伊红（HE）染色、马松（Masson）染色观察心房组织的病理变化及胶原生成情况，并计算心肌胶原体积百分比（CVF），酶联免疫吸附试验（ELISA）法测定血清白细胞介素-1β（IL-1β）水平。确定效果最佳经方后，将30只雄性SD大鼠，随机分为空白组、模型组、真武汤高剂量组、真武汤低剂量组及维拉帕米组，每组6只。空白组尾静脉注射生理盐水，其4组通过连续7 d尾静脉注射Ach-CaCl2构建房颤模型。造模成功后各组予相应药物灌胃14 d。通过Ⅱ导联心电图比较房颤诱发时间与持续时间，HE染色与Masson染色观察心房组织病理变化，ELISA检测血清IL-1β水平，蛋白质印迹（Western blotting）法检测心房组织NLRP3、凋亡相关斑点样蛋白质（ASC）、Caspase-1蛋白表达。结果：真武汤组、维拉帕米组大鼠房颤诱发时间长于模型组，房颤持续时间短于模型组，差异有统计学意义（P<0.01或P<0.05）；炙甘草汤组、桂枝甘草汤组、真武汤组、苓桂术甘汤组、维拉帕米组大鼠CVF低于模型组（P<0.01）；炙甘草汤组、真武汤组、苓桂术甘汤组、维拉帕米组大鼠血清IL-1β水平低于模型组（P<0.01）。真武汤高剂量组、维拉帕米组大鼠房颤诱发时间长于模型组（P<0.05），房颤持续时间短于模型组（P<0.01）；真武汤高剂量组、真武汤低剂量组、维拉帕米组大鼠CVF、血清IL-1β水平均低于模型组（P<0.01）；真武汤高剂量组、真武汤低剂量组、维拉帕米组大鼠心房组织NLRP3、ASC、Caspase-1蛋白相对表达量均低于模型组（P<0.05或P<0.01）。结论：炙甘草汤、桂枝甘草汤、真武汤及苓桂术甘汤均能改善房颤大鼠症状，其中真武汤可以延长房颤诱发时间，缩短房颤持续时间，减轻心房组织病理损伤及纤维化。其作用机制可能与抑制NLRP3/Caspase-1信号通路有关。

关键词：心房颤动；真武汤；炙甘草汤；桂枝甘草汤；苓桂术甘汤；核苷酸结合结构域富含亮氨酸重复序列和含热蛋白结构域受体3/胱天蛋白酶-1通路；炎症反应；大鼠

Abstract：

Objective: To compare four Jingfang [Zhigancao decoction (炙甘草汤), Guizhigancao decoction (桂枝甘草汤), Zhenwu decoction (真武汤), Lingguizhugan decoction (苓桂术甘汤) ] for their effects on rat atrial fibrillation (AF), identify the most effective formulation, and explore its impact on the inflammatory response via the nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3)/Caspase-1 signaling pathway. Methods: A total of 42 male Sprague-Dawley (SD) rats were randomly allocated into blank group, model group, Zhigancao decoction group, Guizhigancao decoction group, Zhenwu decoction group, Lingguizhugan decoction group, and verapamil group, with 6 in each. Rats were injected with physiological saline via tail vein injection in blank group. AF was induced in other 6 groups by daily tail vein injections of an Ach-CaCl2 mixture for seven days. Following model establishment, rats were orally administered the corresponding drug for 14 days. Electrocardiograms were recorded to determine AF initiation time and duration. Hematoxylin-eosin (HE) staining and Masson staining were performed to observe pathological changes and collagen deposition in atrial tissue, respectively. The collagen volume fraction (CVF) was also calculated. Serum interleukin-1β (IL-1β) level was measured by enzyme-linked immunosorbent assay (ELISA). Comparative analysis of experimental data identified the most effective Jingfang. An additional 30 male SD rats were divided into blank group, model group, Zhenwu decoction low dose group, Zhenwu decoction high dose group, and verapamil group, with 6 in each. Rats were injected with physiological saline via tail vein injection in blank group. AF was induced in other 4 groups by daily tail vein injections of an Ach-CaCl2 mixture for seven days. Following model establishment, rats were orally administered the corresponding drug for 14 days. Atrial fibrillation induction time and duration were compared via Lead Ⅱ ECG. Pathological changes in atrial tissue were observed by HE and Masson staining. Serum IL-1β level  was measured by ELISA. The protein expression of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC) and Caspase-1 in atrial tissue was detected by Western blotting. Results: The Zhenwu decoction group and verapamil group exhibited significantly longer AF induction time and shorter AF duration compared to the model group (P<0.01 or P<0.05). Zhigancao decoction group, Guizhigancao decoction group, Zhenwu decoction group, Lingguizhugan decoction group and verapamil group showed lower CVF than model group (P<0.01). Zhigancao decoction group, Zhenwu decoction group, Lingguizhugan decoction group and verapamil group showed lower serum IL-1β level than model group (P<0.01). Zhenwu decoction high dose group and verapamil group exhibited significantly longer AF induction time (P<0.05) and shorter AF duration compared to the model group (P<0.01). Zhenwu decoction high dose group, Zhenwu decoction low dose group and verapamil group showed lower CVF and serum IL-1β level than model group (P<0.01). Zhenwu decoction high dose group, Zhenwu decoction low dose group and verapamil group showed lower protein expression of NLRP3, ASC and Caspase-1 than model group (P<0.05 or P<0.01). Conclusion: Zhigancao decoction, Guizhigancao decoction, Zhenwu decoction and Lingguizhugan decoction can ameliorate AF symptoms in rats. Zhenwu decoction can extend AF initiation time, reduce AF duration, and improve pathological lesions and fibrosis, potentially through suppression of the NLRP3/Caspase-1 signaling pathway.

Key words：atrial fibrillation; Zhenwu decoction; Zhigancao gecoction; Guizhigancao gecoction; Lingguizhugan gecoction; nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3/Caspase-1 signaling pathway; inflammatory reaction; rat

发布时间：2025-11-29

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