Effects of Electro-Acupuncture Based on the Midnight-Noon Ebb-Low Method with Hour-Prescription on BMAL1 Protein and NETs Markers in Lung Tissue of Chronic Obstructive Pulmonary Disease Model Rats with Lung-<italic style="font-style: italic">Qi</italic> Deficiency Syndrome

JIAO-LI Changtian; SHIO Nise Pantaleo; CAI Lianqing; JIANG Qingyao; TANG Siyu; WAN Qianqian; WAN Mengchen; YE Yuqi; ZHU Jie

doi:10.13288/j.11-2166/r.2026.13.012

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Effects of Electro-Acupuncture Based on the Midnight-Noon Ebb-Low Method with Hour-Prescription on BMAL1 Protein and NETs Markers in Lung Tissue of Chronic Obstructive Pulmonary Disease Model Rats with Lung-Qi Deficiency Syndrome

更新时间：2026-06-29

- Effects of Electro-Acupuncture Based on the Midnight-Noon Ebb-Low Method with Hour-Prescription on BMAL1 Protein and NETs Markers in Lung Tissue of Chronic Obstructive Pulmonary Disease Model Rats with Lung-Qi Deficiency Syndrome
  增强出版
- Journal of Traditional Chinese Medicine Vol. 67, Issue 13, Pages: 1422-1430(2026)
- 作者机构：
  
  1.安徽中医药大学中西医结合学院，安徽省合肥市新站区龙子湖路350号，230012
  2.安徽中医药大学新安医学教育部重点实验室
- 作者简介：
- 基金信息：
- DOI：10.13288/j.11-2166/r.2026.13.012
  CLC：
- Received：09 December 2025，
  
  Revised：2026-04-01，
  
  Accepted：18 March 2026，
  
  Published：02 July 2026
- 稿件说明：
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焦李长天,SHIO Nise Pantaleo,蔡炼清等.子午流注纳子法电针对慢性阻塞性肺疾病肺气虚证模型大鼠肺组织BMAL1蛋白及NETs标志物的影响[J].中医杂志,2026,67(13):1422-1430.

JIAO-LI Changtian,SHIO Nise Pantaleo,CAI Lianqing,et al.Effects of Electro-Acupuncture Based on the Midnight-Noon Ebb-Low Method with Hour-Prescription on BMAL1 Protein and NETs Markers in Lung Tissue of Chronic Obstructive Pulmonary Disease Model Rats with Lung-Qi Deficiency Syndrome[J].Journal of Traditional Chinese Medicine,2026,67(13):1422-1430.
焦李长天,SHIO Nise Pantaleo,蔡炼清等.子午流注纳子法电针对慢性阻塞性肺疾病肺气虚证模型大鼠肺组织BMAL1蛋白及NETs标志物的影响[J].中医杂志,2026,67(13):1422-1430. DOI： 10.13288/j.11-2166/r.2026.13.012.

JIAO-LI Changtian,SHIO Nise Pantaleo,CAI Lianqing,et al.Effects of Electro-Acupuncture Based on the Midnight-Noon Ebb-Low Method with Hour-Prescription on BMAL1 Protein and NETs Markers in Lung Tissue of Chronic Obstructive Pulmonary Disease Model Rats with Lung-Qi Deficiency Syndrome[J].Journal of Traditional Chinese Medicine,2026,67(13):1422-1430. DOI： 10.13288/j.11-2166/r.2026.13.012.

摘要

目的

从脑和肌肉芳香烃受体核转运样蛋白1（BMAL1）及中性粒细胞胞外陷阱（NETs）角度探讨子午流注纳子法电针治疗慢性阻塞性肺疾病（COPD）肺气虚证的可能作用机制。

方法

实验分两阶段进行。在疗效观察部分将24只大鼠随机分为对照1组、模型1组、子午电针1组、常规电针组，每组6只。采用烟熏联合气管滴注脂多糖（LPS）法建立COPD肺气虚证大鼠模型。造模成功后，子午电针1组大鼠于卯时（5：00—7：00）取“太渊”“肺俞”“足三里”行电针治疗，每次30 min，隔日1次，持续14天，共7次；常规电针组大鼠采用相同电针处理，于白天随机工作时段（8：00—18：00）施治。检测肺功能指标［包括第0.3秒用力呼气容积（FEV

0.3

）、用力呼气肺活量（FVC）、FEV

0.3

/FVC］，HE染色观察肺组织病理，检测大鼠血浆中炎症因子白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）水平，检测肺组织活性氧（ROS）含量、BMAL1蛋白水平及NETs标志物［髓过氧化物酶（MPO）、中性粒细胞弹性蛋白酶（NE）、瓜氨酸化组蛋白H3（CitH3）］表达。在机制验证部分将36只大鼠分为对照2组、模型2组、子午电针2组、过表达空载组、过表达组、过表达子午电针组，每组6只。过表达组、过表达子午电针组大鼠于造模第5周向气管滴注100 μl过表达

Bmal1

腺病毒载体悬液，过表达空载组大鼠则于相同时间点向肺部注射等滴度、等体积的不包含

Bmal1

基因序列的空载病毒液。子午电针2组和过表达子午电针组电针干预方法同前。检测各组大鼠肺组织MPO、NE、CitH3水平。

结果

与对照1组比较，模型1组FEV

0.3

、FVC、FEV

0.3

/FVC降低，IL-1β、TNF-α水平及ROS含量升高，BMAL1蛋白表达水平下调，MPO、NE、CitH3水平均升高（

＜0.05）；扫描电镜可观察到典型的NETs样结构，纤维网络表面可见少量颗粒状物质；HE染色显示肺泡结构破坏伴炎性浸润。与模型1组比较，子午电针1组与常规电针组上述指标均显著改善，且以子午电针1组改善最显著（

＜0.05）。子午电针1组的NETs样结构减少及肺组织病理改善亦较常规电针组显著。机制验证结果显示，与模型2组比较，子午电针2组、过表达组及过表达子午电针组MPO平均光密度值及MPO、NE、CitH3蛋白水平均显著降低（

＜0.05），且过表达子午电针组低于子午电针2组（

＜0.05）。

结论

子午流注纳子法电针可能通过上调肺组织生物钟蛋白BMAL1表达，抑制NETs在肺组织的过度积累，从而发挥对COPD肺部炎性的改善作用。

Abstract

Objective

To explore the potential mechanism underlying electroacupuncture therapy with the midnight-noon ebb-low method with hour-prescription for chronic obstructive pulmonary disease （COPD） with lung-

deficiency syndrome from the perspective of brain and muscle basic helix-loop-helix ARNT-like 1 （BM

AL1） as well as neutrophil extracellular traps （NETs）.

Methods

The experiment was conducted in two phases. For the therapeutic effectiveness observation， 24 rats were randomly allocated into control group 1， model group 1， midnight-noon electro-acupuncture group 1 and conventional electro-acupuncture group， with 6 rats per group. Rat models of COPD with lung-

deficiency syndrome were established via cigarette smoke exposure combined with intratracheal instillation of lipopolysaccharide （LPS）. After successful modelling， rats in midnight-noon electro-acupuncture group 1 received electro-acupuncture at acupoints "Taiyuan （LU 9）"， "Feishu （BL 13）" and "Zusanli （ST 36）" during 05：00—07：00， 30 minutes per treatment once every other day for 14 consecutive days （7 sessions in total）. Rats in the conventional electro-acupuncture group received identical electroacupuncture manipulation at random daytime hours （08：00—18：00）. Pulmonary function parameters including forced expiratory volume in 0.3 second （FEV

0.3

）， forced vital capacity （FVC） and FEV

0.3

/FVC ratio were detected. Pulmonary histopathological changes were observed via hematoxylin-eosin （HE） staining. Plasma levels of pro-inflammatory factors interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α）， as well as pulmonary reactive oxygen species （ROS） content， BMAL1 protein expression and the levels of neutrophil extracellular traps （NETs） biomarkers such as myeloperoxidase （MPO）， neutrophil elastase （NE）， and citrullinated histone H3 （CitH3） in lung tissue were determined. For the mechanistic verification phase， 36 rats were divided into control group 2， model group 2， midnight-noon electro-acupuncture group 2， overexpression empty group， overexpression BMAL1 group and overexpression with midnight-noon electro-acupuncture group， with 6 rats in each group. At the 5th week of model construction， rats in the overexpression BMAL1 group and overexpression with midnight-noon electro-acupuncture gro

up received intratracheal instillation of 100 μl adenoviral suspension carrying overexpressed

Bmal1

gene. Rats in the overexpression empty group were injected with an equal titer and equal volume of blank adenovirus without the

Bmal1

coding sequence into the lung at the identical time point. Midnight-noon electro-acupuncture group 2 and overexpression with midnight-noon electro-acupuncture group were consistent with the aforementioned protocol. Finally， pulmonary levels of MPO， NE and CitH3 were measured in all groups.

Results

Compared with the control group 1， the model group 1 exhibited decreased FEV

0.3

， FVC and FEV

0.3

/FVC ratio， elevated plasma IL-1β， TNF-α and pulmonary ROS levels， downregulated lung BMAL1 protein expression， and increased contents of MPO， NE and CitH3 （all

＜0.05）； typical NETs-like structures with sparse granular substances attached to fibrous networks were observed under scanning electron microscopy； HE staining revealed damaged alveolar architecture accompanied by inflammatory cell infiltration. Compared with model group 1， both the midnight-noon electro-acupuncture group 1 and conventional electro-acupuncture group achieved obvious improvements in all above indicators， with superior therapeutic effects in midnight-noon electro-acupuncture group 1 （

＜0.05）. Consistently， midnight-noon electro-acupuncture group 1 showed more prominent alleviation of NETs-like structure formation and lung pathological injury compared to the conventional electro-acupuncture group. Mechanistic validation results demonstrated that compared with model group 2， the average optical density of MPO as well as the protein levels of MPO， NE and CitH3 were markedly reduced in midnight-noon electro-acupuncture group 2， overexpression BMAL1 group and overexpression with midnight-noon electro-acupuncture group （

＜0.05）； furthermore， these parameters were significantly lower in the over

expression with midnight-noon electro-acupuncture group than in midnight-noon electro-acupuncture group 2 （

＜0.05）.

Conclusion

Electro-acupuncture based on the midnight-noon ebb-low method with hour-prescription may alleviate pulmonary inflammation in COPD by upregulating the expression of clock protein BMAL1 and inhibiting excessive accumulation of NETs in lung tissue.

关键词

Keywords

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⁰

Effects of Electro-Acupuncture Based on the Midnight-Noon Ebb-Low Method with Hour-Prescription on BMAL1 Protein and NETs Markers in Lung Tissue of Chronic Obstructive Pulmonary Disease Model Rats with Lung-Qi Deficiency Syndrome

DOI：10.13288/j.11-2166/r.2026.13.012

摘要

Abstract

关键词

Keywords

references