凉血活血中药对咪喹莫特诱导小鼠银屑病样皮损模型IL-23/IL-17炎症轴的作用

Effects of blood-cooling and blood-activating medicinal on IL-23/IL-17 axis in psoriasiform mice model induced by imiquimod

目的观察凉血活血中药对咪喹莫特诱导小鼠银屑病样皮损模型白细胞介素-23/白细胞介素-17(IL-23/IL-17)炎症轴的干预作用。方法 BALB/c雄性小鼠随机分为正常组,模型组,凉血活血中药高、中、低剂量组,采用银屑病皮损严重程度指数(PASI)评分观察银屑病样小鼠模型皮损动态变化;光镜观察皮损组织病理变化;采用酶联免疫法(ELISA)、荧光定量PCR、蛋白免疫印迹分析技术检测小鼠血清及皮损中细胞因子含量、mRNA、蛋白的表达水平。结果凉血活血中药高、中、低剂量组小鼠皮肤红斑的出现、色泽及浸润范围、鳞屑面积、鳞屑厚度、皮肤增厚均小于同期的模型组改变(P<0.01或P<0.05)。HE染色显示:凉血活血中药组小鼠皮肤表皮层较平整,角化不全的细胞明显减少,表皮层厚度明显低于模型组(P<0.01)。皮损蛋白及mRNA检测显示,凉血活血中药组小鼠皮肤中IL-17、IL-23蛋白及IL-23mRNA、IL-22mRNA、孤独核受体γt(RORγt)mRNA表达水平均低于模型组(P<0.01或P<0.05)。结论凉血活血中药可抑制咪喹莫特诱导的银屑病样炎症爆发的强度及银屑病样皮损的形成,可能与降低IL-23/IL-17轴相关细胞因子蛋白和mRNA表达水平,并抑制Th17细胞分化转录因子RORγt mRNA的表达有关。

Objective To observe the effects of blood-cooling and blood-activating medicinal（ Liang Xue Huo Xue Yao） on IL-23 / IL-17 axis in psoriasiform mice model induced by imiquimod. Methods BALB / c male mice were randomly divided into control group,model group,and three blood-cooling and bloodactivating medicinal groups of high,medium and low dose（ LXHX-H,LXHX-M and LXHX-L groups）respectively. Pathological changes of skin lesions in mice were observed by using light microscope. The levels,mRNA and protein expression of the cytokines in the serum and skin lesions of mice were tested by using ELISA,RT-PCR,and western blotting assay respectively. Results Compared with model group,LXHX-H,LXHX-M and LXHX-L groups decreased the emergence,color change and infiltration of skin erythema,as well as the PASI scores,epidermal parakeratosis and epidermal over-proliferation of the mice. According to the trend graph of the PASI scores,the area under the curve of LXHX-M group was significantly lower than that in model group. HE staining showed that,compared with model group,mice in LXHX groups had more smooth epidermis cortex,less epidermal parakeratosis and decreased epidermal over-proliferation（ P〈0. 01）. Lesions protein and mRNA test showed that compared with model group,mice in LXHX groups had lower expression level of IL-17 and IL-23 protein,as well as IL-23 mRNA,IL-22 mRNA,and RORγt mRNA（ P〈0. 01 or P〈0. 05）. Conclusion LXHX medicinal could significantly decrease the intensity of the psoriasiform inflammation and the formation of psoriasiform skin lesions induced by imiquimod,the effect of which might be related to the decrease of IL-23 / IL-17 axis cytokine protein and the expression of mRNA,as well as the inhibition of the expression of Th17 cell differentiation transcription factor RORγt mRNA.