益气养阴通络方调控miR-885-5p/PTEN信号通路参与胶原诱导型关节炎模型大鼠细胞自噬-炎症的机制研究  

Mechanism of Yiqi Yangyin Tongluo Decoction(益气养阴通络方)Regulating miR-885-5p/PTEN Signaling Pathway to Regulate Autophagy-Inflammation in Collagen-Induced Arthritis Model Rats

作　　者：张博纶 杨松楠 鞠宝兆[3] 高明利[4] 姜泉[5] 于静[4] 赵夜雨[4] 齐庆[4] 杜立英 ZHANG Bolun;YANG Songnan;JU Baozhao;GAO Mingli;JIANG Quan;YU Jing;ZHAO Yeyu;QI Qing;DU Liying(Shenyang Medical College,Shenyang 110034,Liaoning,China;Shenyang Jianzhong Phrmaceuticd Co.Ltd.,Shenyang 110089,Liaoning,China;Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,Liaoning,China;Guang′anmen Hospital,Chinese Academy of Chinese Medical Sciences,Beijing 100000,China)

机构地区：[1]沈阳医学院,辽宁沈阳110034 [2]沈阳市建中医药有限公司,辽宁沈阳110089 [3]辽宁中医药大学,辽宁沈阳110847 [4]辽宁中医药大学附属医院,辽宁沈阳110032 [5]中国中医科学院广安门医院,北京100000

出　　处：《辽宁中医杂志》2025年第3期182-189,I0004,F0003,共10页Liaoning Journal of Traditional Chinese Medicine

基　　金：国家重点研发计划项目(2018YFC1705202);辽宁省中医药临床学(专)科能力建设项目(2017-32);中医药古籍文献和特色技术传承专项(GZY-KJS-2020-045);《中华医藏》提要编纂项目(二、三编)(KJS-ZHYC-2020-010);辽宁省中央引导地方科技发展专项(2020JH610500004)。

摘　　要：目的通过体内实验探讨益气养阴通络方介导miR-885-5p/磷酸酶和紧张素同源物(PTEN)信号通路参与胶原诱导型关节炎(collagen induced arthritis,CIA)模型大鼠细胞自噬-炎症的作用机制。方法选取40只SD雄性大鼠随机分为4组,每组10只,分别为正常组(CTRL)、模型组(MOD)、益气养阴通络方组(TCM)、甲氨蝶呤组(MTX)。除CTRL组外,其余各组造模后分别接受相应的药物或灭菌注射用水灌胃,给药8周。①评价关节炎症状态:采用关节炎指数(arthritis index,AI)、关节肿胀度直观评价关节炎性状态,苏木精-伊红染色(hematoxylin-eosin,HE)评价关节病理,酶联免疫吸附(enzyme-linked immunosorbent assay,ELISA)法测定血清白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-18(interleukin-18,IL-18)水平评价炎性水平。②评价miR-885-5p/PTEN通路表达情况及细胞自噬表型:采用蛋白免疫印迹(Western blot,WB)检测大鼠滑膜组织PTEN及自噬标志蛋白LC3Ⅱ/Ⅰ、p62蛋白表达情况,实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)测定组织miR-885-5p相对表达水平,透射电镜观察组织细胞自噬小体及溶酶体情况。结果①炎性水平方面:与MOD组比较,TCM组、MTX组均能够降低踝关节肿胀度、AI指数、IL-1β、IL-18水平,差异有统计学意义(P<0.05),同时改善踝关节滑膜组织病理学破坏;TCM与MTX的改善情况差异无统计学意义(P>0.05)。②miR-885-5p/PTEN通路表达情况及细胞自噬表型方面:与MOD组比较,TCM组、MTX组PTEN、LC3Ⅱ/Ⅰ蛋白表达水平提高,miR-885-5p mRNA表达水平提高,p62蛋白表达下降,差异有统计学意义(P<0.05),电镜下自噬小体和溶酶体增多;与MTX组比较,TCM组各项结果相近,差异无统计学意义(P>0.05)。结论miR-885-5p/PTEN信号通路介导的细胞自噬是益气养阴通络方干预CIA大鼠的重要作用机制之一。Objective To explore the mechanism of Yiqi Yangyin Tongluo Decoction(益气养阴通络方)in mediating the miR-885-5p/PTEN signaling pathway involved in autophagy-inflammation of collagen induced arthritis(CIA)model rats through in vivo experiments.Methods Forty male SD rats were randomly divided into four groups(n=10):normal control group(CTRL),model group(MOD),Yiqi Yangyin Tongluo Decoction(TCM)group and methotrexate group(MTX).All groups except the CTRL group were treated with intragastric administration of drugs or sterilized water for injection for 8 weeks after modeling.①Evaluation of joint inflammatory state:The arthritis index(AI)and the degree of joint swelling were used to visually evaluate the arthritic state,while hematoxylin-eosin(HE)staining was used to evaluate the joint pathology.Serum levels of interleukin-1β(IL-1β)and interleukin-18(IL-18)were measured by enzyme-linked immunosorbent assay(ELISA)to evaluate the inflammatory level.②Evaluation of the expression of miR-885-5p/PTEN pathway and autophagy phenotype:Western blot(WB)was used to detect the expression of PTEN and autophagy marker proteins LC3 II/I and p62 in rat synovial tissue,and quantitative real-time PCR(qRT-PCR)was used to determine the relative expression level of miR-885-5p in the tissue.The autophagosomes and lysosomes were observed under the transmission electron microscope.Results①Inflammatory level:Compared with MOD,TCM and MTX could reduce the ankle swelling,AI index,IL-1βand IL-18 with statistical difference(P<0.05),and improve the histopathological damage of ankle synovial membrane.There was no significant difference in improvement between TCM and MTX groups(P>0.05).②The expressions of miR-885-5p/PTEN pathway and autophagy phenotype:Compared with MOD,the expression levels of PTEN and LC3 II/I proteins in TCM and MTX groups were increased,the molecular level of miR-885-5P was increased,and the expression of p62 protein was decreased(P<0.05).The autophagosomes and lysosomes were increased under the electron microsco

关 键 词：益气养阴通络方 类风湿关节炎 细胞自噬 miR-885-5p PTEN信号通路 

分 类 号：R285[医药卫生—中药学]