桑桔四君饮对慢性阻塞性肺疾病大鼠JAK/STAT信号通路介导的炎症和氧化应激及纤维化的影响*

作者：韩萍萍1，孙 烨1，蔡 云1，夏欣欣1，胡 珊1，孙连庆1，刘 昳1，蔡 培2

单位：1.西安交通大学第一附属医院，陕西 西安 710061； 2.西安国际医学中心医院胸科医院，陕西 西安 710119

引用：引用：韩萍萍，孙烨，蔡云，夏欣欣，胡珊，孙连庆，刘昳，蔡培.桑桔四君饮对慢性阻塞性肺疾病大鼠JAK/STAT信号通路介导的炎症和氧化应激及纤维化的影响[J].中医药导报,2025,31(5):20-27.

DOI：10.13862/j.cn43-1446/r.2025.05.004

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摘要：

目的：探究桑桔四君饮（SJSJY）对慢性阻塞性肺疾病（COPD）大鼠Janus激酶（JAK）/信号转导和转录激活子（STAT）信号介导的炎症、氧化应激和纤维化的影响。方法：将72只大鼠随机分为正常组（n=12）与造模组（n=60），造模组大鼠采用烟熏法和脂多糖（LPS）诱导建立COPD大鼠模型。将造模成功的大鼠随机分为模型组、桑桔四君饮低剂量组、桑桔四君饮中剂量组、桑桔四君饮高剂量组及激动剂组，每组12只。正常组和模型组大鼠灌胃生理盐水，桑桔四君饮低、中、高剂量组大鼠分别灌胃0.5、1.0、2.0 g/（kg·d）的桑桔四君饮，激动剂组大鼠灌胃桑桔四君饮[2.0 g/（kg·d）]并腹腔注射1 mg/（kg·d）Colivelin TFA（C-TFA，STAT3激动剂），各组大鼠均给药2周。给药结束后分别检测各组大鼠肺功能、支气管肺泡灌洗液（BALF）中炎症因子水平及肺组织氧化应激指标水平，采用苏木素-伊红（HE）染色和Masson三色染色观察肺形态和纤维化程度，并测量胶原体积分数（CVF），采用Western blotting检测肺组织中α-平滑肌肌动蛋白（α-SMA）、Ⅰ型胶原α1链（COL1A1）、转化生长因子-β1（TGF-β1）、结缔组织生长因子（CTGF）、纤连蛋白（Fibronectin）、磷酸化-Janus激酶2（p-JAK2）和磷酸化-信号转导和转录激活子3（p-STAT3）的蛋白表达水平，采用免疫荧光染色检测肺组织中α-SMA的表达。结果：模型组大鼠肺组织出现明显损伤，最大自主分钟通气量（MVV）、0.3 s用力呼气量（FEV0.3）/用力肺活量（FVC）及最大呼气峰流速（PEF）均低于正常组（P<0.05），CVF高于正常组（P<0.05）；模型组大鼠BALF中白介素-1β（IL-1β）、白介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）及巨噬细胞炎性蛋白-2（MIP-2）水平均高于正常组（P<0.05）；模型组大鼠肺组织超氧化物歧化酶（SOD）及过氧化氢酶（CAT）水平均低于正常组（P<0.05），丙二醛（MDA）水平高于正常组（P<0.05）；模型组大鼠肺组织α-SMA、COL1A1、TGF-β1、CTGF、Fibronectin蛋白相对表达量以及JAK2、STAT3蛋白磷酸化水平均高于正常组（P<0.05）。桑桔四君饮中、高剂量组大鼠肺组织损伤明显减轻，MVV、FEV0.3/FVC及PEF均高于模型组（P<0.05），CVF低于模型组（P<0.05）；桑桔四君饮低、中、高剂量组大鼠BALF中IL-1β、IL-6、TNF-α及MIP-2水平均低于模型组（P<0.05）；桑桔四君饮低、中、高剂量组大鼠肺组织SOD及CAT水平均高于模型组（P<0.05），MDA水平低于模型组（P<0.05）；桑桔四君饮低、中、高剂量组大鼠肺组织α-SMA、COL1A1、TGF-β1、CTGF、Fibronectin蛋白相对表达量以及JAK2、STAT3蛋白磷酸化水平均低于模型组（P<0.05）。激动剂组大鼠肺组织损伤加重，MVV、FEV0.3/FVC及PEF均低于桑桔四君饮高剂量组（P<0.05），CVF高于桑桔四君饮高剂量组（P<0.05）；激动剂组大鼠BALF中IL-1β、IL-6、TNF-α及MIP-2水平均高于桑桔四君饮高剂量组（P<0.05）；激动剂组大鼠肺组织SOD及CAT水平均低于桑桔四君饮高剂量组（P<0.05），MDA水平高于桑桔四君饮高剂量组（P<0.05）；激动剂组大鼠肺组织α-SMA、COL1A1、TGF-β1、CTGF、Fibronectin蛋白相对表达量以及JAK2、STAT3蛋白磷酸化水平均高于桑桔四君饮高剂量组（P<0.05）。结论：桑桔四君饮可有效改善COPD大鼠的肺功能并减轻肺损伤，其机制与抑制JAK2/STAT3信号通路介导的炎症、氧化应激和纤维化有关。

关键词：慢性阻塞性肺疾病；桑桔四君饮；炎症；氧化应激；纤维化；JAK/STAT信号通路；大鼠

Abstract：

Objective: To explore the effects of Sangju Sijun Yin (SJSJY) on Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signal-mediated inflammation, oxidative stress and fibrosis in rats with chronic obstructive pulmonary disease (COPD). Methods: Totally 72 rats were randomly divided into normal group (n=12) and modelling group (n=60). The rats in the modelling group were given smoke and lipopolysaccharide (LPS) induction to establish COPD rat model. The rats that were successfully modeled were randomly divided into model group, SJSJY low dose group, SJSJY medium dose group, SJSJY high dose group and agonist group, with 12 rats in each group. The rats were intragastrically administered with normal saline in normal group and model group. The rats in SJSJY low dose group, SJSJY medium dose group and SJSJY high dose group were intragastrically administered with 0.5, 1.0 and 2.0 g/(kg·d) of SJSJY, respectively. And rats in agonist group were given with 2.0 g/(kg·d) of SJSJY and intraperitoneally injected with 1 mg/(kg·d) Colivelin TFA (C-TFA, STAT3 agonist). All rats were treated for 2 weeks. After the treatment, the lung function test, the levels of inflammatory factors in bronchoalveolar lavage fluid (BALF), and the levels of oxidative stress indicators in lung tissue were measured. Hematoxylin eosin (HE) staining and Masson trichrome staining were performed on lung tissue to observe the lung morphology and degree of fibrosis, and the collagen volume fraction (CVF) was measured. The protein expression levels of α-smooth muscle actin (α-SMA), collagen type I alpha 1 chain (COL1A1), transforming growth factor-β1 (TGF-β1), connective tissue growth factor (CTGF), fibronectin, phosphorylated-Janus kinase 2 (p-JAK2), and phosphorylated-signal transducer and activator of transcription 3 (p-STAT3) in lung tissue were detected by Western blotting. The expression of α-SMA in lung tissue was detected by immunofluorescence staining. Results: The lung tissue of the rats in model group showed obvious damage. The model group showed lower maximal voluntary ventilation (MVV), forced expiratory volume in 0.3 s (FEV0.3)/forced vital capacity (FVC) and peak expiratory flow (PEF) than normal group (P<0.05), while higher CVF than normal group (P<0.05). The model group showed higher levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and macrophage inflammatory protein-2 (MIP-2) in BALF, than normal group (P<0.05). The model group showed lower levels of superoxide dismutase (SOD) and catalase (CAT) in lung tissue, than normal group (P<0.05), while higher level of malondialdehyde (MDA) than normal group (P<0.05). The model group showed higher relative expression levels of α-SMA, COL1A1, TGF-β1, CTGF, fibronectin and the phosphorylation levels of JAK2 and STAT3 in lung tissue, than normal group (P<0.05). The lung tissue damage of rats in SJSJY medium dose group and SJSJY high dose group were significantly alleviated. SJSJY medium dose group and SJSJY high dose group showed higher MVV, FEV0.3/FVC and PEF than model group (P<0.05), while lower CVF than model group (P<0.05). The SJSJY low dose group, SJSJY medium dose group and SJSJY high dose group showed lower levels of IL-1β, IL-6, TNF-α and MIP-2 in BALF, than model group (P<0.05). The SJSJY low dose group, SJSJY medium dose group and SJSJY high dose group showed higher levels of SOD and CAT in lung tissue, than model group (P<0.05), while lower levels of MDA than model group (P<0.05). The SJSJY low dose group, SJSJY medium dose group and SJSJY high dose group showed lower relative expression levels of α-SMA, COL1A1, TGF-β1, CTGF, fibronectin protein and the phosphorylation levels of JAK2 and STAT3 protein in lung tissue, than model group (P<0.05). The lung tissue injury of rats in the agonist group was aggravated. The agonist group showed lower MVV, FEV0.3/FVC and PEF than those SJSJY high dose group (P<0.05), while higher CVF than SJSJY high dose group (P<0.05). The agonist group showed higher levels of IL-1β, IL-6, TNF-α and MIP-2 in BALF, than SJSJY high dose group (P<0.05). The agonist group showed lower levels of SOD and CAT in lung tissue, than SJSJY high dose group (P<0.05), while higher level of MDA than SJSJY high dose group (P<0.05). The agonist group showed higher relative expression of α-SMA, COL1A1, TGF-β1, CTGF, fibronectin proteins and the phosphorylation levels of JAK2 and STAT3 proteins than SJSJY high dose group (P<0.05). Conclusion: Sangju Sijun Yin can effectively improve the lung function and reduce lung damage in COPD rats, and its mechanism is related to the inhibition of JAK2/STAT3 signal-mediated inflammation, oxidative stress and fibrosis.

Key words：chronic obstructive pulmonary disease; Sangju Sijun Yin; inflammation; oxidative stress; fibrosis; JAK/STAT signaling pathway; rat

发布时间：2025-12-23

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