针刺松弛哮喘急性发作大鼠气道平滑肌的作用机制

乔赟; 梁燕娣; 卓少元; 杨雪捷; 梁韬; 凌希; 张倩; 时宜蓉; 易蔚

doi:10.13703/j.0255-2930.20230426-k0004

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针刺松弛哮喘急性发作大鼠气道平滑肌的作用机制

机制探讨 | 更新时间：2024-06-03

- 针刺松弛哮喘急性发作大鼠气道平滑肌的作用机制
- Effect and mechanism of acupuncture on airway smooth muscle relaxation during acute asthma attack in rats
- 中国针灸 2024年44卷第3期页码：295-302
- 作者机构：
  
  1.广西中医药大学，针灸推拿学院，南宁530200
  2.广西中医药大学，基础医学院，南宁530200
  3.广西中医药大学，研究生院，南宁530200
- 作者简介：
  
  易蔚，教授。E-mail：doctoryiwei@126.com
- 基金信息：
  
  2020年广西自然科学基金项目(2020GXNSFAA297021)
- DOI：10.13703/j.0255-2930.20230426-k0004
  中图分类号：
- 收稿：2023-04-26，
  
  网络首发：2024-01-08，
  
  纸质出版：2024-03-12
- 稿件说明：
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乔赟,梁燕娣,卓少元,等.针刺松弛哮喘急性发作大鼠气道平滑肌的作用机制[J].中国针灸,2024,44(3):295-302.

QIAO Yun,LIANG Yandi,ZHUO Shaoyuan,et al.Effect and mechanism of acupuncture on airway smooth muscle relaxation during acute asthma attack in rats[J].Chinese Acupuncture & Moxibustion,2024,44(03):295-302.
乔赟,梁燕娣,卓少元,等.针刺松弛哮喘急性发作大鼠气道平滑肌的作用机制[J].中国针灸,2024,44(3):295-302. DOI： 10.13703/j.0255-2930.20230426-k0004.

QIAO Yun,LIANG Yandi,ZHUO Shaoyuan,et al.Effect and mechanism of acupuncture on airway smooth muscle relaxation during acute asthma attack in rats[J].Chinese Acupuncture & Moxibustion,2024,44(03):295-302. DOI： 10.13703/j.0255-2930.20230426-k0004.

摘要

目的

探讨针刺“肺俞”“定喘”和“孔最”“鱼际”松弛哮喘急性发作大鼠气道平滑肌的作用机制，并比较两组穴位及联合配伍的疗效。

方法

将40只SPF级雄性SD大鼠随机分成空白组、模型组、肺俞定喘组（针刺“肺俞”“定喘”）、孔最鱼际组（针刺“孔最”“鱼际”）和联合组（针刺“肺俞”“定喘”“孔最”“鱼际”），每组8只。除空白组外，其余各组均以卵清蛋白（OVA）联合氢氧化铝凝胶的方法诱导大鼠哮喘急性发作模型。第15天起，除空白组和模型组外，其余各组分别予针刺干预，每次30 min，每日1次，连续14 d。比较各组大鼠的引喘时间，应用肺功能检测仪测定各组大鼠气道阻力（RL）和肺顺应性（Cdyn），ELISA法检测各组大鼠血清及支气管肺泡灌洗液（BALF）内皮素1（ET-1）、肿瘤坏死因子α（TNF-α）、环磷酸腺苷（cAMP）、环磷酸鸟苷（cGMP）含量，Masson染色及透射电镜观察各组大鼠气道平滑肌形态及超微结构，实时荧光定量PCR法检测各组大鼠肺组织ET-1、β

肾上腺素受体（β

-AR）mRNA表达，Western blot法检测各组大鼠肺组织 ET-1、β

-AR蛋白表达。

结果

与空白组比较，模型组大鼠引喘时间缩短（

0.05），不同乙酰甲胆碱浓度（0.025 mg/kg、0.05 mg/kg、0.1 mg/kg、0.2 mg/kg）下RL均升高、Cdyn均降低（

0.05）；与模型组比较，肺俞定喘组、孔最鱼际组及联合组大鼠引喘时间延长（

0.05），不同乙酰甲胆碱浓度下RL均降低、Cdyn均升高（

0.05）；与肺俞定喘组比较，联合组大鼠引喘时间延长（

0.05）。与空白组比较，模型组大鼠血清及BALF ET-1、TNF-α、cGMP含量升高（

0.05），cAMP含量降低（

0.05）；与模型组比较，肺俞定喘组、孔最鱼际组、联合组大鼠血清及BALF ET-1、TNF-α、cGMP含量降低（

0.05），cAMP含量升高（

0.05）。空白组大鼠肺组织结构正常；模型组肺组织可见大量胶原纤维增生，平滑肌增厚，线粒体明显肿胀，嵴大量断裂、减少；孔最鱼际组大鼠肺组织大量胶原纤维增生，平滑肌较空白组增厚，线粒体轻度肿胀，嵴部分断裂；肺俞定喘组、联合组大鼠肺组织改变较模型组明显减轻，线粒体略微肿胀，嵴少量断裂。与空白组比较，模型组大鼠肺组织ET-1 mRNA及蛋白表达升高、β

-AR mRNA及蛋白表达降低（

0.05）；与模型组比较，肺俞定喘组、孔最鱼际组、联合组大鼠肺组织ET-1 mRNA及蛋白表达降低、β

-AR mRNA及蛋白表达升高（

0.05）；与肺俞定喘组比较，联合组大鼠肺组织β

-AR mRNA表达升高（

0.05）；与孔最鱼际组比较，联合组大鼠肺组织β

-AR mRNA表达升高、ET-1蛋白表达降低（

0.05）。

结论

针刺“肺俞”“定喘”、“孔最”“鱼际”及针刺两组穴位均可缓解哮喘急性发作大鼠的气道平滑肌痉挛，其作用机制可能是抑制ET-1 mRNA及蛋白表达，使ET-1、TNF-α分泌减少；同时升高β

-AR mRNA及蛋白表达，平衡cAMP和cGMP含量。针刺两组穴位疗效更佳。

Abstract

Objective

To explore the effect and mechanism of acupuncture at "Feishu" (BL 13) and "Dingchuan" (EX-B 1)

and "Kongzui" (LU 6) and "Yuji" (LU 10) for relaxing the airway smooth muscle in the rats during acute asthma attack and compare the effect among the two pairs of acupoints and the acupoints combination.

Methods

Forty SD male rats with SPF grade were randomly divided into a blank group

a model group

a pair-point A group (acupuncture at "Feishu" [BL 13

]

and "Dingchuan" [EX-B 1

]

)

a pair-point B group (acupuncture at "Kongzui" [LU 6

]

and "Yuji" [LU 10

]

) and a point combination group (acupuncture at "Feishu" [BL 13

]

"Dingchuan" [EX-B 1

]

"Kongzui" [LU 6

]

and "Yuji" [LU 10

]

)

with 8 rats in each group. Except the rats in the blank group

the model of acute asthma attack was induced by ovalbumin (OVA) combined with aluminum hydroxide gel in the rest groups. Started on the 15th day of modeling

except in the blank group and the model group

acupuncture was delivered in the other groups

30 min in each intervention

once daily

for 14 days. In each group

the latent period of asthma inducing was measured; the lung resistance (LR) and dynamic lung compliance (Cdyn) were determined using lung function detector; the levels of endothelin-1 (ET-1)

tumor necrosis factor-α (TNF-α)

cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) in serum and bronchoalveolar lavage fluid (BALF) were

measured by ELISA; with Masson staining and electron microscopy adopted

the morphology and ultrastructure of airway smooth muscle of the rats were observed; the mRNA and protein expressions of ET-1 and beta-2 adrenergic receptor

(β

-AR) were detected by quantitative real-time fluorescence and Western blot

respectively.

Results

Compared with the blank group

the latent period of asthma inducing was shortened (

0.05)

RL increased and Cdyn decreased (

0.05) with the different concentrations of methacholine (0.025 mg/kg

0.05 mg/kg

0.1 mg/kg

0.2 mg/kg) in the model group. In the pair-point A group

the pair-point B group and the point combination group

the latent period of asthma inducing was prolonged (

0.05)

RL decreased and Cdyn increased (

0.05) with different concentrations of methacholine when compared with those in the model group; and the latent period of asthma inducing in the point combination group was longer than that in the pair-point A group (

0.05). Compared with the blank group

the levels of ET-1

TNF-α and cGMP in the serum and BALF were elevated (

0.05)

and those of cAMP reduced (

0.05) in the model group. The levels of ET-1

TNF-α and cGMP in the serum and BALF were reduced (

0.05)

and those of cAMP elevated (

0.05) in the pair-point A group

the pair-point B group and the point combination group when compared with those in the model group. In the blank group

the lung tissue was normal structurally. In the model group

the collagen fibers were proliferated increasingly

the smooth muscle was thickened

the mitochondria were swollen

and their cristae disrupted and reduced massively. In the pair-point B group

the collagen fibers were proliferated

the smooth muscle was thicker compared with that in the blank group

the mitochondria were mildly swol

len and their cristae disrupted partially. In the pair-point A group and the point combination group

the lung tissue changes were obviously alleviated in comparison with the model group

the mitochondria were slightly swollen and their cristae disrupted occasionally. Compared with the blank group

the mRNA and protein expression of ET-1 increased and that of β

-AR decreased in the lung tissue of the model group (

0.05). In the pair-point A group

the pair-point B group and the point combination group

the mRNA and protein expression of ET-1 was reduced and that of β

-AR elevated in the lung tissue when compared with those in the model group (

0.05). In comparison with the pair-point A group

the mRNA expression of β

-AR was elevated in the point combination group (

0.05). When compared with the pair-point B group

the mRNA expression of β

-AR increased

the protein expression of ET-1 decreased (

0.05) in the point combination group.

Conclusion

Acupuncture at "Feishu" (BL 13) and "Dingchuan" (EX-B 1)

"Kongzui" (LU 6) and "Yuji" (LU 10)

two pairs of acupoints relieves the airway smooth muscle spasm in the rats during acute asthma attack

which may be related to inhibiting the mRNA and protein expression of ET-1 to reduce the excretion of ET-1 and TNF-α; while enhancing the mRNA and protein expression of β

-AR to balance the levels of cAMP and cGMP. The effect is optimal when acupuncture is delivered at two pairs of acupoints simultaneously.

关键词

Keywords

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