升麻鳖甲汤对银屑病小鼠表皮炎症及角质形成细胞增殖的影响

The Effect of Shengma Biejia Decoction(升麻鳖甲汤)on Epidermal Inflammation and Proliferation of Keratinocytes in Psoriasis Mice

目的:探讨升麻鳖甲汤对银屑病小鼠表皮炎症及角质形成细胞增殖的影响及机制。方法:将48只BALB/c小鼠随机分为空白组、模型组、甲氨蝶呤组、升麻鳖甲汤低剂量组、升麻鳖甲汤中剂量组、升麻鳖甲汤高剂量组,每组8只。除空白组外,其余各组采用咪喹莫特造模并予相应药物干预,观察小鼠银屑病皮损严重程度指数(PASI)评分;干预结束后处死小鼠,取小鼠皮肤HE染色观察皮损部病理变化并评定Baker评分;采用蛋白质印迹法(Western blotting)检测小鼠皮损部p-STAT3、STAT3蛋白相对表达量;采用酶联免疫吸附法(ELISA)测定皮损部白细胞介素-17(IL-17)、IL-22、维甲酸相关孤儿核受体-γt(ROR-γt)蛋白含量;采用免疫组化法检测小鼠皮损部Ki-67的阳性细胞表达率。结果:模型组小鼠PASI评分、Baker评分均高于空白组(P<0.01);升麻鳖甲汤中剂量组、升麻鳖甲汤高剂量组、甲氨蝶呤组小鼠PASI评分、Baker法评分均低于模型组(P<0.01)。模型组小鼠皮损部p-STAT3/STAT3高于空白组(P<0.05);升麻鳖甲汤高剂量组、甲氨蝶呤组小鼠皮损部p-STAT3/STAT3均低于模型组(P<0.05)。模型组小鼠皮损部IL-17、IL-22、ROR-γt蛋白含量及Ki-67阳性细胞率高于空白组(P<0.01);甲氨蝶呤组小鼠皮损部IL-17、IL-22、ROR-γt蛋白含量及Ki-67阳性细胞率低于模型组(P<0.01);升麻鳖甲汤高剂量组小鼠皮损部IL-17蛋白含量低于模型组(P<0.01);升麻鳖甲汤中、高剂量组小鼠皮损部IL-22、ROR-γt蛋白含量及Ki-67阳性细胞率均低于模型组(P<0.01)。结论:升麻鳖甲汤可能通过抑制STAT3信号通路活性,抑制皮损部IL-17、IL-22、ROR-γt分泌及角质形成细胞增殖,发挥治疗银屑病的作用。

Objective:To study the effect and mechanism of Shengma Biejia decoction on epidermal inflammation and keratinocyte proliferation in psoriasis mice.Method:Totally 48 BALB/c mice were divided into blank group,model group,methotrexate group,Shengma Biejia decoction low dose group(low dose group),Shengma Biejia decoction medium dose group(medium dose group),and Shengma Biejia decoction high dose group(high dose group),with 8 mice in each group.Except blank group,all other groups were modeled with imiquimod and intervened with corresponding drugs.The Psoriasis Area and Severity Index(PASI)score were observed in mice.After the intervention,the mice were euthanized.And their skin was stained with HE to observe the pathological changes in the skin lesions and scored using the Baker method.Western blotting was used to detect the expression levels of STAT3 and p-STAT3 proteins in skin lesions.Enzyme linked immunosorbent assay(ELISA)was used to determine IL-17,IL-22 and ROR-γt in the skin lesions.Immunohistochemical method was used to detect the expression rate of Ki-67 positive cells in skin lesions.Results:The model group showed higher PASI score and Baker method score than blank group(P<0.01).The medium dose group,high dose group and methotrexate group showed lower PASI score and Baker method score than model group(P<0.01).The model group showed higher p-STAT3/STAT3 than blank group(P<0.05).The high dose group and methotrexate group showed lower p-STAT3/STAT3 than model group(P<0.05).The model group showed higher IL-17,IL-22 and ROR-γt protein expression and expression rate of Ki-67 positive cells than blank group(P<0.01).The methotrexate group showed lower IL-17,IL-22 and ROR-γt protein expression and expression rate of Ki-67 positive cells than model group(P<0.01).The high dose group showed lower IL-17 protein expression than model group(P<0.01).The medium dose group and high dose group showed lower IL-22 and ROR-γt protein expression and expression rate of Ki-67 positive cells than model group(P<0.01).Conclusion:Shengma Biejia decoction may inhibit the activity of STAT3 signaling pathway and inhibit IL-17,IL-22,and RORγt in skin lesions secretion and proliferation of keratinocytes,to play a therapeutic role in psoriasis.