白鲜碱对特应性皮炎小鼠PKCα/NF-κB通路及肥大细胞浸润的影响  

Effects of dictamnine on PKCα/NF-κB pathway and mast cell infiltration in mice with atopic dermatitis

作　　者：张春华 马欣[2] 丁佩军[2] 彭明霞 ZHANG Chun-hua;MA Xin;DING Pei-jun;PENG Ming-xia(Department of Dermatology,Shanghai Baoshan Hospital of Integrated Traditional Chinese and Western Medicine(Baoshan Branch,Shuguang Hospital,Shanghai University of Traditional Chinese Medicine),Shanghai 201999;Department of Dermatology,Shuguang Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海市宝山区中西医结合医院皮肤科(上海中医药大学附属曙光医院宝山分院皮肤科),上海201999 [2]上海中医药大学附属曙光医院皮肤科,上海201203

出　　处：《热带医学杂志》2021年第6期711-716,739,F0004,共8页Journal of Tropical Medicine

基　　金：上海市卫生健康委员会名老中医学术经验研究工作室建设项目(SHGZS-2017020);上海市卫生健康委员会临床研究专项项目(20184Y0020);上海市卫生健康委员会海派中医流派传承人才培养项目(LPRC2017041)。

摘　　要：目的探讨白鲜碱对激活蛋白激酶Cα(PKCα)/核转录因子κB(NF-κB)通路的调控作用,及对特应性皮炎(AD)小鼠肥大细胞浸润的影响。方法按照随机数字表法将C57BL/6小鼠分为正常对照组、模型组、白鲜碱低剂量(1.2 mg/kg)组、白鲜碱中剂量(2.4 mg/kg)组、白鲜碱高剂量(4.8 mg/kg)组、地塞米松组(阳性对照),每组12只。除正常对照组外,其他组小鼠用2,4-二硝基氯苯(DNCB)反复刺激建立AD模型。白鲜碱低、中、高剂量组小鼠背部每天分别涂抹1.2、2.4、4.8 mg/kg白鲜碱,地塞米松组涂抹地塞米松乳膏,模型组及正常对照组涂抹10mL/kg橄榄油,各组连续给药7 d,1次/d。观察小鼠抓搔次数及背部皮损程度;ELISA法检测血清组胺及免疫球蛋白E(IgE)水平;HE及甲苯胺蓝染色检测背部皮损组织病理变化及肥大细胞浸润情况;类胰蛋白酶免疫组化法检测肥大细胞活化状态;WB法测定PKCα、NF-κBp65及磷酸化NF-κBp65(p-NF-κBp65)、酪氨酸激酶(Lyn)及磷酸化Lyn(p-Lyn)、脾酪氨酸激酶(Syk)及磷酸化Syk(p-Syk)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、IL-1β等蛋白表达。结果与正常对照组比较,模型组小鼠背部皮肤角化、增厚及炎性损伤较重,血清组胺及IgE水平、搔抓次数、肥大细胞浸润、PKCα、p-NF-κBp65/NF-κBp65、p-Lyn/Lyn、p-Syk/Syk、TNF-α、IL-6、IL-1β表达升高,差异有统计学意义(P<0.05)。与模型组比较,白鲜碱低、中、高剂量组和地塞米松组小鼠背部损伤缓解,血清组胺及IgE水平、搔抓次数、肥大细胞浸润、PKCα、p-NF-κBp65/NF-κBp65、p-Lyn/Lyn、p-Syk/Syk、TNF-α、IL-6、IL-1β表达降低,差异有统计学意义(P<0.05);且呈剂量依赖性。白鲜碱高剂量组与地塞米松组比较差异无统计学意义(P>0.05)。结论白鲜碱可能通过抑制PKCα/NF-κBp65通路活化,降低肥大细胞浸润活化介导的炎症反应,来改善AD小鼠炎症及瘙痒症状。Objective To investigate the regulatory effect of dictamnine on activated protein kinase Cα(PKCα)/nuclear transcription factor kappa B(NF-κB)pathway and its influence on mast cell infiltration in atopic dermatitis(AD)mice.Methods C57 BL/6 mice were divided into normal control group,model group,low dose(1.2 mg/kg)dictamnine group,medium dose(2.4 mg/kg)dictamnine group,high dose(4.8 mg/kg)dictamnine group and dexamethasone group(positive control)by random number table,with 12 in each group. Except for the normal control group,mice in other groups were repeatedly stimulated with 2,4-dinitrochlorobenzene(DNCB)to establish AD models. The backs of mice in the low,medium and high dose dictamnine group were smeared with 1.2,2.4,4.8 mg/kg dictamnine per day;the dexamethasone group applied dexamethasone cream;the model group and the normal control group applied 10 mL/kg olive oil,each group was continuously administered for 7 days,once a day. The scratching times and the degree of skin lesions on the back of mice in each group were observed;the levels of histamine and immunoglobulin E(IgE)in serum were detected by ELISA method;the pathological changes and mast cell infiltration of the skin lesions were detected by HE and toluidine blue staining;and the activation status of mast cells was detected by tryptase immunohistochemical staining;the expression of PKCα,NF-κBp65 and phosphorylated NF-κBp65(p-NF-κBp65),tyrosine kinase(Lyn)and phosphorylated Lyn(pLyn),spleen tyrosine kinase(Syk)and phosphorylated Syk(p-Syk),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-1β(IL-1β)proteins was detected by Western blotting(WB). Results Compared with the normal control group,the keratinization,thickening and inflammatory injury of the back skin in the model group were serious,the serum histamine and IgE levels,scratching times,mast cell infiltration,PKCα,p-NF-κBp65/NF-κBp65,p-Lyn/Lyn,p-Syk/Syk,TNF-α,IL-6,IL-1β expression levels were increased,and the differences were statistically significant(P<0.05).Compare

关 键 词：白鲜碱 蛋白激酶CΑ 核转录因子ΚB 特应性皮炎 肥大细胞浸润 

分 类 号：R758.32[医药卫生—皮肤病学与性病学]