SHP2调控巨噬细胞表型对系统性硬皮病纤维化病变的影响  

Effects of SHP2 regulating macrophage phenotypic transformation on fibrotic lesions in systemic scleroderma

作　　者：龙阳华 王珍真 LONG Yang-Hua;WANG Zhen-zhen(Department of Pharmacy,Xiangyang Central Hospital,Affiliated Hospital of Hubei University of Arts and Sciences,Xiangyang 441000,Hubei Province,China)

机构地区：[1]湖北文理学院附属医院、襄阳市中心医院药剂科,湖北襄阳441000

出　　处：《中国临床药理学杂志》2024年第2期244-248,共5页The Chinese Journal of Clinical Pharmacology

基　　金：湖北省知识创新专项(自然科学基金)基金资助项目(2019CFB448)。

摘　　要：目的探究含Src同源2结构域蛋白酪氨酸磷酸酶(SHP2)对系统性硬皮病(SSc)小鼠炎性反应及纤维化病变的影响,及其作用机制。方法将40只小鼠随机分为空白组、模型组和低、高剂量实验组,每组10只。除空白组外,其余3组小鼠均皮下注射博来霉素建立SSc模型。造模成功后,低、高剂量实验组分别腹腔注射2.5和5.0 mg·kg^(-1)PHPS1;空白组和模型组均腹腔注射等量的0.9%NaCl。4组小鼠每天给药1次,连续给药20 d。用免疫荧光染色法检测巨噬细胞浸润情况,用流式细胞术测定巨噬细胞极化情况,用实时荧光定量聚合酶链反应法检测M1型巨噬细胞肿瘤坏死因子-α(TNF-α)、诱导型一氧化氮合酶(iNOS)与M2型巨噬细胞重组人精氨酸酶1(Arg-1)、转化生长因子-β(TGF-β)的表达水平。结果高剂量实验组和模型组、空白组的外周血F4/80+CD86+标记的M1型巨噬细胞比例分别为(16.81±1.59)%、(36.95±3.54)%和(1.70±0.15)%,F4/80+CD206+标记的M2型巨噬细胞比例分别为(18.49±1.76)%、(2.99±0.27)%和(16.62±1.49)%;上述3组皮肤组织中巨噬细胞标记物F4/80相对荧光强度分别为1.46±0.13、2.14±0.19和1.00±0.04,TNF-αmRNA分别为2.09±0.20、3.54±0.34和1.00±0.05,iNOS mRNA分别为1.63±0.15、3.28±0.31和1.00±0.04,Arg-1 mRNA分别为0.75±0.07、0.38±0.03和1.00±0.06,TGF-βmRNA分别为0.79±0.08、0.41±0.04和1.00±0.04;上述3组肺组织中TNF-αmRNA分别为1.46±0.14、2.61±0.25和1.00±0.04,iNOS mRNA分别为1.53±0.15、2.82±0.29和1.00±0.03,Arg-1 mRNA分别为0.67±0.07、0.31±0.03和1.00±0.05,TGF-βmRNA分别为0.71±0.07、0.39±0.04和1.00±0.05。高剂量实验组的上述指标与模型组相比较,在统计学上差异均有统计学意义(均P<0.05)。结论SHP2抑制剂作用于SSc小鼠能够改善其皮肤组织和肺组织的炎性损伤及纤维化,该作用与调控巨噬细胞极化相关。Objective To investigate the effect of Src homology 2 domain-containing protein tyrosine phosphatase(SHP2)on inflammatory response and fibrosis in mice with systemic scleroderma(SSc)and its mechanism.Methods Forty mice were randomly divided into blank group,model group and experimental-L,-H groups,with 10 mice in each group.Except the blank group,the other three groups of mice were established SSc model.The experimental-L,-H groups we re given PHPS1 treatment of 2.5 mg·kg^(-1)and 5 mg·kg^(-1),respectively;blank group and model group were intraperitoneally injected with 0.9%NaCl.Four groups of mice were given the drug once a day for 20 days.Immunofluorescence staining was used to detect the macrophage infiltration.Flow cytometry was used to determine the macrophage polarization,real-time quantitative fluorescence polymerase chain reaction was used to detect the expression of M1 macrophage markers tumor necrosis factor-α(TNF-α),inducible nitric oxide synthase(iNOS)and M2 macrophage markers recombinant human arginase 1(Arg-1),transforming growth factor-β(TGF-β)in skin tissues and lung tissues.Results The proportions of M1-type macrophages labeled F4/80+CD86+in peripheral blood of experimental-H group,model group,blank group were(16.81±1.59)%,(36.95±3.54)%,(1.70±0.15)%,respectively;the proportions of M2-type macrophages labeled F4/80+CD206+were(18.49±1.76)%,(2.99±0.27)%,(16.62±1.49)%,respectively.The relative fluorescence intensity of F4/80 of macrophage marker in skin tissue were 1.46±0.13,2.14±0.19,1.00±0.04,respectively;TNF-αmRNA were 2.09±0.20,3.54±0.34,1.00±0.05,respectively;iNOS mRNA were 1.63±0.15,3.28±0.31,1.00±0.04,respectively,Arg-1 mRNA were 0.75±0.07,0.38±0.03,1.00±0.06,respectively,TGF-βmRNA were 0.79±0.08,0.41±0.04,1.00±0.04,respectively.In lung tissue,TNF-αmRNA were 1.46±0.14,2.61±0.25,1.00±0.04,respectively;iNOS mRNA were 1.53±0.15,2.82±0.29,1.00±0.03,respectively,Arg-1 mRNA were 0.67±0.07,0.31±0.03,1.00±0.05,respectively;TGF-βmRNA were 0.71±0.07,0.39±0.04,1.0

关 键 词：系统性硬皮病 含Src同源2结构域蛋白酪氨酸磷酸酶 炎症 组织纤维化 巨噬细胞极化 肺损伤 

分 类 号：R97[医药卫生—药品]