摘要
目的:基于Toll样受体4(TLR4)/髓分化因子88(MyD88)/核转录因子-κB(NF-κB)信号通路探讨清热利湿中药对痤疮的治疗作用及机制。方法:制备大鼠胃肠湿热证耳廓复合痤疮模型,分为对照组,模型组,清热利湿方高、低剂量组(13.5、4.5 g·kg^(–1)),连续给药14 d。造模21 d后观察大鼠耳廓形态及厚度变化,苏木素-伊红(HE)染色观察病理变化,原位末端转移酶标记(TUNEL)染色检测细胞凋亡率,透射电镜观察棘细胞亚显微结构,酶联免疫吸附分析(ELISA)检测大鼠血清白细胞介素-6(IL-6)、IL-1β和肿瘤坏死因子-α(TNF-α)水平,蛋白质免疫印迹法(Western blot)检测大鼠耳组织TLR4、MyD88、NF-κB、BCL2相关蛋白X(Bax)、B淋巴细胞瘤-2(Bcl-2)的蛋白质表达水平。结果:与对照组相比,模型组大鼠耳廓出现红肿、增厚、变硬、质地干燥粗糙、组织角化、表皮增厚、棘细胞出现凋亡现象,耳组织中TLR4、MyD88、NF-κB、Bax的蛋白质表达水平显著升高,Bcl-2的蛋白质表达量显著降低(P<0.05);与模型组相比,清热利湿方组大鼠耳廓痤疮样皮损明显好转,病理损伤减轻,角质层和棘层厚度减少,细胞凋亡率降低,血清炎症因子水平下降,组织中TLR4、MyD88、NF-κB、Bax的蛋白质表达水平明显降低,Bcl-2的蛋白质表达水平明显升高。结论:清热利湿中药可能通过抑制TLR4/MyD88/NF-κB信号通路发挥抗炎、抗凋亡作用,进而治疗痤疮。
Objective:To explore the therapeutic effect and mechanism of Qingre Lishi Prescription on acne based on the Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)nuclear factor kappa-B(NF-κB)signaling pathway.Methods:A rat model of auricular composite acne with the syndrome of dampness-heat in stomach and intestine was established.Rats were grouped as follows:blank,model,high and low doses(13.5、4.5 g·kg–1,respectively)of Qingre Lishi Prescription and administrated with corresponding agents for 14 d.After 21 d of modeling,the morphological and thickness changes of rat auricles were observed.Hematoxylin-eosin staining was used to observe pathological changes,and terminal deoxynucleotidyl transferase-mediated nick end labeling was employed to detect the apoptosis rate of cells.Transmission electron microscopy was employed to observe the submicroscopic structure of heckle cells.Enzyme-linked immunosorbent assay was employed to measure the levels of interleukin-6(IL-6),interleukin-1β(IL-1β),and tumor necrosis factor-α(TNF-α)in the serum.Western blot was employed to determine the protein levels of TLR4,MyD88,NF-κB,B-cell lymphoma-2(Bcl-2),and Bcl-2-associated X protein(Bax)and in the rat ear tissue.Results:Compared with those in the blank group,the auricles of rats in the model group showed redness,swelling,thickening,hardening,dry and rough texture,keratinization,epidermal thickening,and apoptosis of heckle cells.In addition,the ear tissue in the model group presented up-regulated protein levels of TLR4,MyD88,NF-κB,and Bax and down-regulated protein level of Bcl-2(P<0.05).Compared with the model group,the Qingre Lishi Prescription groups demonstrated reduced acne-like skin lesions of the auricles,alleviated pathological damage,decreased thickness of stratum corneum and spinous layer,lowered apoptosis rate,declined serum levels of inflammatory factors,significantly down-regulated protein levels of TLR4,MyD88,NF-κB,and Bax,and significantly up-regulated protein level of Bcl-2.Conclusion:Qingre Lishi Prescription may play anti-inflammatory and anti-apoptosis roles by inhibiting the TLR4/MyD88/NF-κB signaling pathway,thus treating acne.
作者
孙玉雯
程莹莹
尚津锋
张晓璐
王伯洪
史启今
高琛
金敏
陈雯
SUN Yuwen;CHENG Yingyinga;SHANG Jinfeng;ZHANG Xiaolu;WANG Bohong;SHI Qijin;GAO Chen;JIN Min;CHEN Wen(Guang'anmen Hospital,China Academy of Chinese Medical Sciences,Beijing 100053,China;School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China)
出处
《中国现代中药》
2025年第7期1291-1299,共9页
Modern Chinese Medicine
基金
国家中医药管理局2022中医药科学技术研究专项(GZY-KJS-2022-004)
国家中医药管理局中药炮制技术传承基地项目(国中医药科技中药〔2022〕136号)
国家中医药管理局全国老药工传承工作室建设项目(国中医药人教函〔2024〕255号)
中央高水平中医医院临床研究和成果转化能力提升项目——医疗机构制剂研发与新药转化专项(HLCMHPP2023056)。
关键词
清热利湿中药
痤疮
炎症
凋亡
Toll样受体4/髓分化因子88/核转录因子-κB信号通路
Qingren Lishi Prescription
acne
inflammation
apoptosis
Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor kappa-B(NF-κB)signaling pathway
