加味荆芩汤对玫瑰痤疮模型小鼠皮损组织炎症及免疫反应的影响  

Effects of Jiawei Jingqin Decoction(加味荆芩汤)on Inflammatory and Immune Responses in Rosacea Model Mice

作　　者：杨瑾[1] 赵丽娟[1] 伍迪[1] 张旭[1] 叶建州[1] YANG Jin;ZHAO Lijuan;WU Di;ZHANG Xu;YE Jianzhou(Yunnan Provincial Hospital of Traditional Chinese Medicine,Kunming,650021)

机构地区：[1]云南省中医医院,云南省昆明市650021

出　　处：《中医杂志》2024年第3期308-316,共9页Journal of Traditional Chinese Medicine

基　　金：国家中医药管理局第七批全国老中医药专家学术经验继承工作项目(国中医药人教函[2022]76号);国家中医药管理局全国名老中医药专家传承工作室建设项目;云南省高层次中医药后备人才培养对象(云卫中医发展发[2021]1号);云南省科学技术厅中医联合专项(202101AZ070001-216)。

摘　　要：目的探讨加味荆芩汤治疗玫瑰痤疮的可能作用机制。方法50只8周龄雌性BALB/c小鼠随机分为正常对照组、模型组和加味荆芩汤低、中、高剂量组,每组10只。除正常对照组外,其余各组小鼠予背部注射40μl抗菌肽LL-37(浓度为320μmol/L),每12 h重复注射1次,共注射4次,以制备玫瑰痤疮小鼠模型,正常对照组小鼠同时间点背部皮内注射40μl PBS溶液。造模成功后加味荆芩汤低、中、高剂量组小鼠分别给予加味荆芩汤4.35、8.71、17.42 g/(kg·d)灌胃,正常对照组和模型组小鼠给予20 ml/(kg·d)生理盐水灌胃。各组均每日1次,持续5天。观察各组小鼠皮损变化,并进行红斑面积测定及红斑程度评分;取注射部位皮肤组织HE染色观察病理形态学变化,甲苯胺蓝染色检测肥大细胞数量;ELISA法检测皮损组织中白细胞介素6(IL-6)、白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)水平;RT-qPCR法检测玫瑰痤疮发病机制中固有免疫关键因子[Toll样受体2(TLR2)及下游分子丝氨酸蛋白酶激肽释放酶5(KLK5)、基质金属蛋白酶9(MMP-9)]与相关炎症因子[IL-1β、IL-6、TNF-α]mRNA,适应性免疫CD4^(+)T细胞相关因子干扰素γ(INF-γ)mRNA表达;免疫组化染色法检测KLK5、MMP-9蛋白表达及CD4^(+)T阳性细胞数;Western blot法检测CD4^(+)T细胞相关因子[IFN-γ、CC趋化因子受体5(CCR5)、信号转导转录激活因子1(STAT1)、信号转导转录激活因子3(STAT3)]蛋白表达水平。结果正常对照组小鼠背部注射部位皮肤正常,未见皮损及红斑,病理显示未见炎性细胞浸润。模型组小鼠红斑程度评分及红斑面积较正常对照组显著升高(P<0.05),病理显示大量炎细胞浸润,部分可见深层结缔组织损伤。与模型组比较,加味荆芩汤中、高剂量组小鼠红斑程度评分及红斑面积均显著降低(P<0.05),炎性渗出及炎性细胞浸润有所减少;加味荆芩汤低剂量组差异无统计学意义(P>0.05),炎细胞浸�Objective To investigate the potential mechanisms of Jiawei Jingqin Decoction(加味荆芩汤)in the treatment of rosacea.Methods Fifty 8-week-old female BALB/c mice were randomly divided into normal control group,model group,and Jiawei Jingqin Decoction low,medium,and high-dose groups,with 10 mice in each group.Except for the normal control group,mice in the other groups were injected with 40μl of antimicrobial peptide LL-37 at a concentration of 320μmol/L on the back,repeated once every 12 hours for a total of 4 injections,to induce the ro⁃sacea mouse model.The mice in the normal control group were injected with 40μl of PBS solution at the same time.After successful modeling,the mice in the Jiawei Jingqin Decoction low,medium,and high-dose groups were orally administered Jiawei Jingqin Decoction at doses of 4.35,8.71,and 17.42 g/(kg·d)respectively,while the mice in the normal control group and the model group were orally administered 20 ml(/kg·d)of normal saline.All groups were treated once daily for 5 days.The changes in skin lesions were observed,and the erythema area was measured and scored.Skin tissue at the injection site was collected for histopathological examination using HE staining,and the number of mast cells was detected using toluidine blue staining.The levels of interleukin-6(IL-6),interleukin-1β(IL-1β),and tumor necrosis factor-alpha(TNF-α)in the skin lesion tissues were measured using the ELISA method.The mRNA expression levels of key factors in the innate immune response pathway of rosacea disease mechanism,including Toll-like receptor 2(TLR2),downstream molecules serine protease kallikrein 5(KLK5)and matrix metal⁃loproteinase 9(MMP-9),as well as related inflammatory factors(IL-1β,IL-6,TNF-α),and adaptive immune CD4^(+)T cell-related factors interferon-gamma(INF-γ)mRNA were detected using RT-qPCR.Immunohistochemical stain⁃ing was performed to detect the protein expression of KLK5,MMP-9,and the number of CD4^(+)T-positive cells.Western blot analysis was used to determine the pro

关 键 词：玫瑰痤疮 CD4^(+)T细胞 炎症因子 免疫反应 加味荆芩汤 

分 类 号：R285.5[医药卫生—中药学]