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[电针预处理通过抑制炎症反应和上调脂多糖诱导的急性肺损伤大鼠中ACE2和Ang (1-7)的表达来改善肺功能]

[Electroacupuncture preconditioning improves pulmonary function via inhibiting inflammatory response and up-regulating expression of ACE2 and Ang (1-7) in lipopolysaccharide-induced acute lung injury rats].

作者信息

Liu Xin-Yue, Su Jing-Chao, Zhang Xin-Fang, Xiang Shui-Ying, Yi Chun-Mei, Wang Yu-Wen, Wang Cai-Yun, Liu Zi-Bing

机构信息

Institute of Acupuncture-moxibustion and Meridian of College of Acupuncture-moxibustion and Massage, Anhui University of Chinese Medicine, Hefei 230038, China.

College of Integrated Traditional Chinese and Western Medicine, Anhui University of Chinese Medicine, Hefei 230038, China.

出版信息

Zhen Ci Yan Jiu. 2022 Aug 25;47(8):684-9. doi: 10.13702/j.1000-0607.20210979.

Abstract

OBJECTIVE

To observe the effect of electroacupuncture (EA) at "Zusanli"(ST36) pretreatment on lung functions, inflammatory response, and levels of angiotensin-converting enzyme 2 (ACE2) and angiotensin (1-7) [Ang (1-7)] in rats with sepsis-induced acute lung injury (ALI), so as to explore its mechanisms underlying improvement of ALI.

METHODS

Thirty male SD rats were randomly divided into normal, model and EA groups (=10 in each group). The sepsis-related ALI model was established by intraperitoneal injection of lipopolysaccharide (LPS, 5 mg/kg). Rats of the EA group received EA (4 Hz/20 Hz, 1-3 mA) stimulation at bilateral ST36 for 30 min, once each day, for 7 days before modeling. The lung functions including forced vital capacity (FVC), forced expiratory volume at 0.1 second (FEV0.1) and FEV0.3 were detected using a respiratory function detector for small animals at 3 h after modeling. The bronchoalveolar lavage fluid (BALF) was collected for assaying the contents of Ang (1-7), tumor necrosis factor-α (TNF-α) and interleukin-1 β (IL-1β) using ELISA. The lung wet/dry weight (W/D) ratio, FEV0.1/FVC, and FEV0.3/FVC were calculated. The histopathological changes of lung tissues were displayed by hematoxylin-eosin (H.E.) staining. The expression of ACE2 and mitochondrial assembly receptor (MasR) mRNAs and proteins in the lung tissue was detected by fluorescence quantitative real-time PCR and Western blot, separately.

RESULTS

Following modeling, the levels of FVC, FEV0.1, FEV0.3, ratio of FEV0.1/FVC and FEV0.3/FVC, content of Ang (1-7) in the BALF, and the expression levels of ACE2 and MasR mRNAs and proteins in the lung tissue were significantly decreased (<0.01), while the level of W/D ratio and TNF-α and IL-1β contents in the BALF significantly increased (<0.01) in the model group relevant to the normal group. In comparison with the model group, the levels of FVC, FEV0.1, FEV0.3, ratio of FEV0.1/FVC and FEV0.3/FVC, content of Ang (1-7) in the BALF, and expression levels of ACE2 and MasR mRNAs and proteins in the lung tissue were significantly increased (<0.05, <0.01), whereas the level of W/D ratio, and TNF-α and IL-1β contents in the BALF were significantly decreased (<0.05, <0.01) in the EA group. H.E. staining showed pulmonary interstitial edema and alveolar septum thickening with severe inflammatory cell infiltration in the model group, which was relatively milder in the EA group.

CONCLUSION

EA preconditioning at ST36 can improve pulmonary function in sepsis-related ALI rats, which may be related to its effects in inhibiting inflammatory response and up-regulating ACE2 and MasR expression and Ang (1-7) content in the lung tissue.

摘要

目的

观察电针“足三里”(ST36)预处理对脓毒症诱导的急性肺损伤(ALI)大鼠肺功能、炎症反应及血管紧张素转换酶2(ACE2)和血管紧张素(1-7)[Ang(1-7)]水平的影响,以探讨其改善ALI的作用机制。

方法

将30只雄性SD大鼠随机分为正常组、模型组和电针组(每组10只)。通过腹腔注射脂多糖(LPS,5 mg/kg)建立脓毒症相关性ALI模型。电针组大鼠在造模前7天,每天1次,给予双侧ST36电针刺激(4 Hz/20 Hz,1-3 mA)30分钟。造模后3小时,使用小动物呼吸功能检测仪检测肺功能,包括用力肺活量(FVC)、0.1秒用力呼气量(FEV0.1)和FEV0.3。收集支气管肺泡灌洗液(BALF),采用酶联免疫吸附测定法(ELISA)检测Ang(1-7)、肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)含量。计算肺湿/干重(W/D)比值、FEV0.1/FVC和FEV0.3/FVC。采用苏木精-伊红(H.E.)染色观察肺组织病理变化。分别采用荧光定量实时聚合酶链反应(PCR)和蛋白质免疫印迹法检测肺组织中ACE2和线粒体装配受体(MasR)的mRNA和蛋白表达。

结果

造模后,模型组大鼠的FVC、FEV0.1、FEV0.3、FEV0.1/FVC和FEV0.3/FVC水平、BALF中Ang(1-7)含量以及肺组织中ACE2和MasR的mRNA和蛋白表达水平均显著降低(<0.01),而W/D比值以及BALF中TNF-α和IL-1β含量显著升高(<0.01),与正常组相比差异有统计学意义。与模型组相比,电针组大鼠的FVC、FEV0.1、FEV0.3、FEV0.1/FVC和FEV0.3/FVC水平、BALF中Ang(1-7)含量以及肺组织中ACE2和MasR的mRNA和蛋白表达水平均显著升高(<0.05,<0.01),而W/D比值以及BALF中TNF-α和IL-1β含量显著降低(<0.05,<0.01)。H.E.染色显示,模型组出现肺间质水肿和肺泡间隔增厚,伴有严重的炎症细胞浸润,电针组病变相对较轻。

结论

ST36电针预处理可改善脓毒症相关性ALI大鼠的肺功能,其机制可能与抑制炎症反应、上调肺组织中ACE2和MasR表达以及Ang(1-7)含量有关。

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