当归芍药散加味调控JNK/p38 MAPK通路改善痤疮炎症和凋亡的机制  

Mechanism of Modified Danggui Shaoyaosan in Improving Inflammation and Apoptosis in Acne via Regulating JNK/p38 MAPK Pathway

作　　者：陈功珍 杨雨齐 刘鑫 唐挺 CHEN Gongzhen;YANG Yuqi;LIU Xin;TANG Ting(Guizhou University of Traditional Chinese Medicine(TCM),Guiyang 550025,China;The First Affiliated Hospital of Guizhou University of TCM,Guiyang 550001,China)

机构地区：[1]贵州中医药大学,贵阳550025 [2]贵州中医药大学第一附属医院,贵阳550001

出　　处：《中国实验方剂学杂志》2025年第5期31-40,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82360929,82260824);贵州省科技计划项目(黔科合基础-ZK[2022]一般501);贵阳中医一附院新进博士启动基金项目(GYZYYFY-BS-2021[03])。

摘　　要：目的:基于c-Jun氨基末端激酶(JNK)和p38丝裂原活化蛋白激酶(p38 MAPK)通路探讨当归芍药散加味对痤疮的治疗作用及机制。方法:SD大鼠建立大鼠耳痤疮模型,分为正常组,模型组,当归芍药散加味低、中、高剂量组(7.15、14.30、28.60 g·kg·d^(-1)),每组6只,连续给药14 d。干预结束后观察大鼠耳廓形态变化,使用苏木素-伊红(HE)染色观察大鼠痤疮耳组织的病理变化,酶联免疫吸附测定法(ELISA)检测大鼠耳组织白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)水平,实时荧光定量聚合酶链式反应(Real-time PCR)检测大鼠耳组织胱天蛋白酶-3(Caspase-3)、B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、JNK和p38 MAPK mRNA水平,蛋白免疫印迹法(Western blot)检测大鼠耳组织Caspase-3、Bcl-2、Bax、JNK和p38 MAPK的蛋白水平。通过网络药理学分析获取当归芍药散加味在治疗痤疮中发挥主要作用的活性成分和关键靶点,并使用分子对接技术对主要活性成分和靶点进行对接。结果:动物实验结果表明,与正常组比较,模型组大鼠耳廓出现红肿、增厚、变硬、质地干燥粗糙,表面见皮脂鳞屑和脱痂,伴有丘疹样隆起或囊肿的痤疮样皮损,组织出现角化、表皮增厚、真皮胶原纤维变性和坏死、皮下肌肉变性和坏死、炎症细胞浸润及纤维组织增生等病理改变,组织中IL-1β、TNF-α、Caspase-3、Bax、JNK和p38 MAPK mRNA和蛋白的表达水平显著升高(P<0.01),Bcl-2 mRNA和蛋白的水平显著降低(P<0.01);与模型组比较,当归芍药散加味各给药组大鼠耳廓痤疮样皮损明显好转,组织病理损害得到不同程度的减轻,组织中IL-1β、TNF-α、Caspase-3、Bax、JNK和p38 MAPK mRNA和蛋白的表达水平明显降低(P<0.05,P<0.01),Bcl-2 mRNA和蛋白的水平明显升高(P<0.05,P<0.01)。网络药理学分析表明,当归芍药散加味在治疗痤疮中发挥主要作用的化合物主要为刺槐素、山柰酚、木犀草素、槲Objective:To explore the therapeutic effects and mechanisms of modified Danggui Shaoyaosan on acne based on the c-Jun N-terminal kinase(JNK)/p38 mitogen-activated protein kinase(p38 MAPK)pathway.Methods:A rat ear acne model was established in SD rats,and the rats were divided into a blank group,a model group,and low-,medium-,and high-dose groups of modified Danggui Shaoyaosan(7.15,14.30,28.60 g·kg·d^(-1)),with six rats in each group.After the administration for 14 consecutive days,morphological changes in the rats'auricles were observed,and hematoxylin-eosin(HE)staining was used to examine the pathological changes in the acne-affected ear tissue.Enzyme-linked immunosorbent assay(ELISA)was employed to measure the levels of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in the ear tissue.Quantitative reverse transcription polymerase chain reaction(Real-time PCR)was performed to detect the mRNA expression levels of Caspase-3,B cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),JNK,and p38 MAPK in the ear tissue.Additionally,Western blot analysis was conducted to assess the protein levels of Caspase-3,Bcl-2,Bax,JNK,and p38 MAPK in the ear tissue.The active components and key targets of modified Danggui Shaoyaosan in the treatment of acne were identified through network pharmacology analysis.Molecular docking was then employed to evaluate the interactions between the main active components and the key targets.Results:The results of the animal experiment demonstrated that compared with those in the blank group,rats in the model group exhibited redness,swelling,thickening,hardening,dryness,and roughness of the auricle.The surface showed sebaceous scales and desquamation,accompanied by acne-like lesions such as papule-like elevations or cysts.Histopathological changes included keratinization,epidermal thickening,dermal collagen fiber degeneration and necrosis,subcutaneous muscle degeneration and necrosis,inflammatory cell infiltration,and fibrous tissue proliferation.The mRNA and protein expression leve

关 键 词：痤疮 当归芍药散加味 c-Jun氨基末端激酶(JNK)和p38丝裂原活化蛋白激酶(p38 MAPK)信号通路 炎症 凋亡 

分 类 号：R285[医药卫生—中药学] R289[医药卫生—中医学] R275