基于p38 MAPK/NF-κB信号通路探讨麻杏苦甘汤对油酸诱导的急性肺损伤大鼠炎症反应及细胞凋亡的影响  

Impact of Maxing Kugan Decoction on Inflammatory Response and Apoptosis in Oleic Acid-induced Acute Lung Injury in Rats via p38 MAPK/NF-κB Signaling Pathway

作　　者：焦太强 南一[2,3] 袁玲[4] 李佳庆 牛阳[2,3] JIAO Taiqiang;NAN Yi;YUAN Ling;LI Jiaqing;NIU Yang(School of Traditional Chinese Medicine,Ningxia Medical University,Yinchuan 750004,China;Ningxia Regional Key Laboratory of Ministry of Education of Traditional Chinese Medicine for Prevention and Treatment of Regional High Incidence Disease,Ningxia Medical University,Yinchuan 750004,China;Ningxia Regional Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Regional High Incidence Disease,Ningxia Medical University,Yinchuan 750004,China;College of Pharmacy,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学中医学院,银川750004 [2]宁夏医科大学宁夏区域高发病中医药防治教育部重点实验室,银川750004 [3]宁夏医科大学宁夏区域高发病中西医结合防治研究重点实验室,银川750004 [4]宁夏医科大学药学院,银川750004

出　　处：《中国实验方剂学杂志》2025年第7期108-116,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：宁夏医科大学校级重点项目(XZ2023010);国家自然科学基金区域创新发展联合基金项目(U20A20404);国家名老中医药专家传承工作室建设项目(中医药教育函〔2022〕75号)。

摘　　要：目的:观察麻杏苦甘汤对油酸(OA)诱导的急性肺损伤(ALI)大鼠炎症反应及细胞凋亡的影响,并初步探讨其作用机制。方法:将60只SD大鼠随机分为正常组,模型组,地塞米松组(2 mg·kg^(−1)),麻杏苦甘汤低、中、高(3.1、6.2、12.4 g·kg^(−1))剂量组。各组分别给予同体积生理盐水和相应浓度麻杏苦甘汤灌胃,连续给药7 d后,模型组和各给药组采用尾静脉注射OA(0.2 mL·kg^(−1))的方法建立ALI模型。造模12 h后,分析各组大鼠血气情况和测定肺组织湿/干质量比(W/D);酶联免疫吸附测定法(ELISA)检测大鼠肺泡灌洗液(BALF)中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)的含量;苏木素-伊红(HE)和原位末端标记法(TUNEL)染色观察肺组织细胞损伤、凋亡情况并对其进行评分;蛋白免疫印迹法(Western blot)和实时荧光定量聚合酶链式反应(Real-time PCR)检测各组大鼠肺组织p38丝裂原活化蛋白激酶/核转录因子-κB(p38 MAPK/NF-κB)信号通路相关和凋亡相关蛋白及mRNA的表达。结果:与正常组比较,模型组大鼠可见氧分压(PaO_(2))、血氧饱和度(SaO_(2))、氧合指数(PaO_(2)/FiO_(2))显著降低,二氧化碳分压(PaCO_(2))、W/D显著升高(P<0.01);BALF中TNF-α、IL-6、IL-1β含量显著升高(P<0.01);肺组织病理染色可见明显炎性浸润、组织水肿、肺泡间隔增厚及肺组织细胞凋亡;肺组织p38 MAPK、NF-κB p65、NF-κB抑制蛋白α(IκBα)、B细胞淋巴瘤-2(Bcl-2)相关X蛋白(Bax)、Caspase-3 mRNA表达和磷酸化(p)-p38 MAPK、p-NF-κB p65、p-IκBα、Bax、胱天蛋白酶-3(Caspase-3)、剪切的(cleaved)Caspase-3蛋白表达显著增高,Bcl-2 mRNA和蛋白表达显著降低(P<0.01);与模型组比较,给予麻杏苦甘汤干预能显著升高大鼠PaO_(2)、SaO_(2)、PaO_(2)/FiO_(2),显著降低PaCO_(2)、W/D(P<0.01);显著降低BALF中TNF-α、IL-6、IL-1β含量(P<0.01);缓解肺组织中炎性浸润、组织水肿、肺泡间隔增厚及肺�Objective:To investigate the effects of Maxing Kugan decoction(MKD)on inflammatory response and apoptosis in rats with oleic acid(OA)-induced acute lung injury(ALI)and explore its mechanism of action.Methods:Sixty Sprague-Dawley(SD)rats were randomly assigned into six groups:a control group,a model group,a dexamethasone-treated group(2 mg·kg^(−1)),and three MKD-treated groups at low,medium,and high doses(3.1,6.2,12.4 g·kg^(−1)).Each group was administered either an equivalent volume of normal saline or the corresponding concentration of MKD by gavage for seven consecutive days.The model group and each administration group were used to establish the ALI model by tail vein injection of OA(0.2 mL·kg^(−1)).Twelve hours after modeling,blood gas analyses were conducted,and the wet-to-dry(W/D)weight ratio of lung tissue was measured for each group.Additionally,enzyme-linked immunosorbent assay(ELISA)was employed to quantify the levels of tumor necrosis factor-alpha(TNF-α),interleukin-1β(IL-1β),and interleukin-6(IL-6)in the bronchoalveolar lavage fluid(BALF)of the rats.Cell damage and apoptosis in lung tissue were examined via hematoxylin-eosin(HE)staining and TdTmediated dUTP-biotin nick end labeling(TUNEL)assays,and the results were subsequently scored.The expression levels of the p38 mitogen-activated protein kinase(p38 MAPK)/nuclear factor kappa-B(NF-κB)signaling pathway and apoptosis-related proteins and mRNAs were assessed using Western blot and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR).Results:Compared with the control group,the model group exhibited a significant decrease in partial pressure of oxygen(PaO_(2)),blood oxygen saturation(SaO_(2)),and oxygenation index(PaO_(2)/FiO_(2)),along with a marked increase in partial pressure of carbon dioxide(PaCO_(2))and lung W/D ratio(P<0.01).Additionally,levels of TNF-α,IL-6,and IL-1βin BALF were significantly elevated(P<0.01).Histopathological analysis of lung tissue showed significant inflammatory infiltration,tissue edem

关 键 词：麻杏苦甘汤 p38丝裂原活化蛋白激酶/核转录因子-κB(p38 MAPK/NF-κB)信号通路 油酸 急性肺损伤 

分 类 号：R289[医药卫生—方剂学] R285[医药卫生—中药学] R259[医药卫生—中医学]