搜风顺气丸对咪喹莫特诱导的银屑病样小鼠γδT17的抑制作用

Inhibition by Soufengshunqi decoction towards γδT17 of psoriatic mice which induced by imiquimod

目的观察搜风顺气丸对咪喹莫特诱导的银屑病样小鼠模型皮损的影响。方法40只雄性BALB/c小鼠,背部备皮后,采用数字表法随机分为空白对照组、模型组、搜风顺气丸组和甲氨蝶呤组,背部涂抹5%(质量分数)咪喹莫特乳膏诱导银屑病样模型。采用银屑病皮损面积和疾病严重程度评分(psoriasis area and severity index,PASI)标准进行评分,测量表皮层厚度,记录形态学变化;免疫荧光染色观察小鼠皮损中Ki-67的表达;免疫组织化学检测皮损CD3+T淋巴细胞的表达;流式细胞术检测胸腺及双侧腹股沟淋巴结CD3+T细胞中CD4+、CD8+、γδT+及RORγt+γδT+细胞比例;Western blotting法检测小鼠皮损中磷酸化信号转导与转录激活因子3(phosphorylated-signal transduction and activators of transcription 3,p-STAT3)蛋白的表达水平。结果与模型组相比,搜风顺气丸组小鼠皮损PASI评分及表皮厚度均明显降低、脾指数(P<0.01),胸腺指数及淋巴结指数(P<0.01)均明显低于模型组、Ki-67阳性细胞表达个数减少(P<0.01);真皮CD3+T细胞表达均低于模型组(P<0.01);双侧腹股沟淋巴结及胸腺CD3+T细胞中CD4+T细胞比例差异无统计学意义,但γδT+及RORγt+γδT+细胞比例均明显减少;并且搜风顺气丸组小鼠皮损中p-STAT3表达下降(P<0.01)。结论搜风顺气丸可能通过抑制RORγt的表达作用,从而减少IL-17A的分泌,改善皮损,表明搜风顺气丸对炎性因子的抑制作用可能是其治疗银屑病的机制之一。

Objective To observe the effect of Soufengshunqi(SFSQ)decoction on the skin lesions induced by imiquimod(IMQ)in silver-like mice,and to explore its mechanism of action in psoriasis.Methods Forty BALB/c mice were shaved on the back.They were randomly divided into control group,IMQ group,Sofengshunqi decoction group and methotrexate(MTX)group by 5%(mass fraction)imiquimod.The back of the special cream is applied to induce a skin psoriasis-like IMQ.The lesion area and severity index(PASI)were observed and scored.Morphological changes and epidermal layer thickness of the lesions were observed under light microscope.Ki-67 was observed by immunofluorescence staining in mouse skin lesions.CD3+T lymphocyte expression in the dermis of the lesions was detected by immunohistochemistry.The ratio of CD4+,CD8+,γδT+and RORγt+γδT+cells in thymus and lymph node CD3+T cells were detected by Western blotting.Expression of the phosphorylated-signal transduction and activators of transcription 3(p-STAT3)was tested.Results Compared with the IMQ group,the PASI score and epidermal thickness of the skin of the SFSQ decoction group were significantly decreased,the spleen index(P<0.01),and the thymus index and lymph node index(P<0.01)were significantly lower than the IMQ group.The number of Ki-67 postive cells decreased(P<0.01).The expression of dermal CD3+T cells was lower than that of the IMQ group(P<0.01).There was no significant difference in CD4+T cells between lymph nodes and thymus CD3+T cells butγδT+and the proportion of RORγt+γδT+cells was significantly reduced.The expression of STAT3 related pathway protein P-STAT3 was decreased in the skin of SFSQ decoction group(P<0.01).Conclusion SFSQ Decoction may inhibit the secretion of IL-17A by inhibiting the expression of RORγt,thereby improving the change of psoriasis-like lesions induced by imiquimod in mice,suggesting the inflammatory response and related factors of SFSQ decoction.Its inhibitory effect may be one of its potential mechanisms for the treatment of psoriasis.