川芎嗪通过miR-143-3p靶向调节LIMK1/cofilin通路减轻脂多糖诱导的小鼠急性肺损伤  

Tetramethylpyrazine attenuates lipopolysaccharide-induced acute lung injury in mice by targeting the LIMK1/cofilin pathway through miR-143-3p

作　　者：薛蕾 刘小碣 陶伟婷 闵思敏 高可飞 李言 张永 XUE Lei;LIU Xiaojie;TAO Weiting;MIN Simin;GAO Kefei;LI Yan;ZHANG Yong(Department of Pulmonary and Critical Care Medicine,the First Affiliated Hospital of Bengbu Medical University,Bengbu,Anhui 233000,China;Anhui Clinical and Preclinical Key Laboratory of Respiratory Disease,Bengbu,Anhui 233000,China;Cardiovascular and Cerebrovascular Diseases Research Center,Bengbu,Anhui 233000,China;Changzhou University,Changzhou,Jiangsu 213000,China)

机构地区：[1]蚌埠医科大学第一附属医院呼吸与危重症医学科,安徽蚌埠233000 [2]安徽省呼吸系病临床基础省级重点实验室,安徽蚌埠233000 [3]心血管疾病基础与临床重点实验室,安徽蚌埠233000 [4]常州大学,江苏常州213000

出　　处：《临床肺科杂志》2025年第1期13-23,共11页Journal of Clinical Pulmonary Medicine

基　　金：国家自然基金资助项目(No.81673791)。

摘　　要：目的 探究川芎嗪(TMP)通过miR-143-3p靶向调节LIM激酶1/丝切蛋白(LIMK1/cofilin)信号通路对脂多糖(LPS)诱导的小鼠急性肺损伤(ALI)的影响。方法 将C57BL/6小鼠随机分为对照组(CON组)、模型组(LPS组)、TMP组、空载病毒组(GFP组)、过表达miR-143-3p组、敲低miR-143-3p组,每组10只。HE染色观察肺组织病理变化情况;ELISA检测肺泡灌洗液(BALF)中白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)含量;肺湿/干比(肺W/D)、BALF中白细胞数量、蛋白浓度以及埃文斯蓝(Evans Blue)染料检测肺血管通透性;Western Blot检测p-LIMK1、p-cofilin的表达情况。结果 与CON组相比,LPS组肺组织病理损伤加重,肺损伤评分、肺W/D、BALF白细胞数量、蛋白浓度和炎症因子、Evans Blue渗漏、p-LIMK1、p-cofilin表达水平增加(P均<0.01);与LPS组相比,过表达miR-143-3p组肺组织病理损伤减轻,肺损伤评分、BALF白细胞数量、蛋白浓度和炎症因子、Evans Blue渗漏降低(P均<0.01),肺W/D降低(P<0.05),p-LIMK1、p-cofilin表达水平降低(P<0.01,P<0.05);与LPS组相比,TMP组肺组织病理损伤减轻,肺损伤评分、肺W/D、BALF白细胞数量、蛋白浓度和炎症因子、Evans Blue渗漏降低(P均<0.01),p-LIMK1、p-cofilin表达水平降低(P<0.01,P<0.05);与TMP组相比,敲低miR-143-3p组逆转了TMP对LPS诱导的小鼠ALI的改善作用(P均<0.01)。双荧光素酶实验证实miR-143-3p可靶向调控LIMK1表达(P<0.001)。结论 TMP通过上调miR-143-3p来抑制LIMK1/cofilin信号通路,从而减轻LPS诱导的小鼠ALI。Objective To investigate the effect of tetramethylpyrazine(TMP)on lipopolysaccharide(LPS)-induced acute lung injury(ALI)in mice through miR-143-3p targeting and regulation of LIM kinase 1/filament cutin(LIMK1/cofilin)signaling pathway.Methods C57BL/6 mice were randomly divided into a control group(CON group),model group(LPS group),TMP group,null virus group(GFP group),overexpression of miR-143-3p group,and knockdown of miR-143-3p group,with 10 mice in each group.HE staining was used to observe the pathological changes of the lung histology;ELISA was used to detect the levels of interleukin 1β(IL-1β),interleukin 6(IL-6),and tumor necrosis factorα(TNF-α)in bronchoalveolar lavage fluid(BALF);lung wet/dry ratio(lung W/D),leukocyte count in BALF,protein concentration in BALF,and Evans Blue dye were used to detect lung vascular permeability;Western blot was used to detect the expression of p-LIMK1 and p-cofilin.Results Compared with the CON group,lung histopathological injury was aggravated in the LPS group,and the expression levels of lung injury score, lung W/D, BALF leukocyte counts, protein concentration and inflammatory factors, Evans Blue leakage, p-LIMK1, and p-cofilin were increased ( P <0.01);compared with the LPS group, lung histopathological injury in the group overexpressing miR-143-3p was attenuated, lung injury score, BALF leukocyte count, protein concentration and inflammatory factors, Evans Blue leakage were decreased (all P <0.01), lung W/D was decreased ( P <0.05), and p-LIMK1, p-cofilin expression levels were decreased ( P <0.01, P <0.05);compared with the LPS group, lung histopathological injury was attenuated in the TMP group, lung injury score, lung W/D, BALF leukocyte count, protein concentration and inflammatory factors, Evans Blue leakage were reduced (all P <0.01), and the expression levels of p-LIMK1, p-cofilin were reduced ( P <0.01, P <0.05);compared with the TMP group, the knockdown of miR-143-3p group reversed the effect of TMP on LPS-induced mouse ALI amelioration (all P <0.01). Dual

关 键 词：川芎嗪 急性肺损伤 LIMK1/cofilin通路 miR-143-3p 

分 类 号：R285.5[医药卫生—中药学]